|Year : 2022 | Volume
| Issue : 77 | Page : 193-200
Elucidating the immunomodulatory effect of daidzein in Benzo(a)pyrene -Induced lung cancer mice model through modulation of proliferating cell nuclear antigen, NF-κB, CYP1A1, and NRF
Jianyu Feng1, Wei Meng2, Yandong Liu1, Cuiyun Li3, Chunnan Zhang1, Peng Wang1, Hesham S Almoallim4, Velu Manikandan5, Hui Guan1
1 Department of Integrated TCM and Western Medicine, Heilongjiang Provincial Hospital, Harbin, Heilongjiang, 150001, China
2 Department of Emergency, Heilongjiang Provincial Hospital, Harbin, Heilongjiang, 150001, China
3 Department of Rehabilitation, Heilongjiang Provincial Hospital, Harbin, Heilongjiang, 150001, China
4 Department of Oral and Maxillofacial Surgery, College of Dentistry, King Saud University, Riyadh -11545, Saudi Arabia
5 Division of Biotechnology, College of Environmental and Bioresource Sciences, Jeonbuk National University, Iksan 54596, South Korea
|Date of Submission||17-Jul-2021|
|Date of Decision||23-Sep-2021|
|Date of Acceptance||14-Dec-2021|
|Date of Web Publication||28-Mar-2022|
Department Western and Medicine and Integrated TCM, Heilongjiang Provincial Hospital, Harbin 150001, Heilongjiang
Source of Support: None, Conflict of Interest: None
| Abstract|| |
Background: Lung cancer is the second most predominant reason for cancer deaths globally. It is estimated to be approximately 30% among the cancer deaths. Daidzein (DAZ) is a polyphenolic compound present commonly in soy-based plants and is proven to have various therapeutic properties. Objectives: In this study, we aimed to understand the immunomodulatory activity of DAZ in the mice model with benzo(a)pyrene (B(a)P)-induced lung carcinoma. Materials and Methods: The mice were divided into five groups: Group I served was the control group; Group II animals were challenged with B(a)P; Group III animals were treated with DAZ before challenge with B(a)P; Group IV animals were treated with DAZ after challenging the animals with B(a)P; and Group V animals were treated with DAZ alone till the end of the experimental period. Tumor incidence was calculated, and the following parameters were analyzed: Body weight, lung weight, total number of tumors, percentage of inhibition, immunoglobulin (Ig) levels (immunoglobulin G, immunoglobulin A, and immunoglobulin M), key marker enzymes, and proinflammatory cytokines in both treated and normal mice. The lung tissues were analyzed through the histopathological analysis. Results: According to our results, all the markers that favor the growth of cancer were increased in the lung cancer group. After the administration of DAZ, all the markers returned to their original levels. Conclusion: In conclusion, DAZ protected the cells against the B(a)P-induced inflammatory responses in lung cancer.
Keywords: Benzo(a)pyrene, carcinoembryonic antigen, CYP1A1, daidzein, lung cancer, tumor necrosis factor-alpha
|How to cite this article:|
Feng J, Meng W, Liu Y, Li C, Zhang C, Wang P, Almoallim HS, Manikandan V, Guan H. Elucidating the immunomodulatory effect of daidzein in Benzo(a)pyrene -Induced lung cancer mice model through modulation of proliferating cell nuclear antigen, NF-κB, CYP1A1, and NRF. Phcog Mag 2022;18:193-200
|How to cite this URL:|
Feng J, Meng W, Liu Y, Li C, Zhang C, Wang P, Almoallim HS, Manikandan V, Guan H. Elucidating the immunomodulatory effect of daidzein in Benzo(a)pyrene -Induced lung cancer mice model through modulation of proliferating cell nuclear antigen, NF-κB, CYP1A1, and NRF. Phcog Mag [serial online] 2022 [cited 2022 Sep 27];18:193-200. Available from: http://www.phcog.com/text.asp?2022/18/77/193/341066
- Lung cancer is known to be one of the most prevalent cancers which cause higher mortality globally
- DAZ initiation in B(a)P-induced lung cancer modulates the antioxidant activities and immune responses of the animals
- DAZ might possess the ability to alter the responses of immune cells, either scavenge or inhibit the formation of reactive oxygen species.
Abbreviations used: DAZ: Daidzein; PAHs: Polycyclic aromatic hydrocarbons; B(a)P: Benzo(a) pyrene; ICDH -Isocitrate dehydrogenase; KDH: α-Ketoglutarate dehydrogenase; SDH: Succinate dehydrogenase; MDH: Malate dehydrogenase.
| Introduction|| |
Worldwide, lung cancer is known to be one of the leading causes of cancer deaths. Statistical reports describe that approximately 2 lakh new lung cancer cases were found including both male and female, among which approximately more than 1 lakh deaths have been reported in 2020., Previous studies have shown that diet plays a major role in initiating the process of cancer., It has been reported that soy food helps in minimizing the risk of lung cancer, especially aggressive cancers.,,
The most frequently used therapeutic strategies are surgery, radiotherapy, and chemotherapy. Although modern chemotherapy is effective during the initial days of treatment, the long duration of chemotherapy can lead to chemoresistance, which is the major reason for treatment failure. Therefore, there is an increasing demand to identify novel drugs with minimal risk for the treatment of cancer. Tobacco smoking is the leading cause of lung cancer, which is caused mainly due to the presence of polycyclic aromatic hydrocarbons (PAHs). The major component having the carcinogenic property of PAHs is benzo(a)pyrene (B(a)P). It is present in the great quantities in cigarettes. One of the key roles played by B(a)P is the formation of DNA adduct, which in turn leads to the initiation of tumor formation. However, an imbalance in the processes of detoxification and metabolic activation processes increases the risk of cancer in an individual., Plant-based compounds initiate various signaling pathways and thus help in inhibiting the damages caused by the carcinogens.
Daidzein (DAZ), also called as 7-hydroxy-3-4-hydroxyphenyl chromen-4-one, is a polyphenolic compound derived from various soy-based plants. Previous studies suggest that DAZ is associated with the isoflavones group shows antioxidative, antidysrhythmic, and anti-inflammatory properties., Studies also show that isoflavone minimize the risk of cancer., In addition, previous reports state that DAZ induces cell death in different types of cancer cells., Many experiments have proven that DAZ inhibits the growth of tumor cells by initiating apoptosis., However, to the best of our knowledge, there are no studies conducted to test the beneficial effects of DAZ against lung cancer. Therefore, in this study, we aimed to discover the therapeutic activity of DAZ against the B(a)P-induced lung cancer in mice.
| Materials and Methods|| |
In this study, healthy male Swiss albino mice (aged from 6 to 10 weeks) weighing 20–30 g were employed. Animals were purchased from the Institutional Animal Facility and were maintained in a 12 h/12 h light–dark cycle. The temperature and humidity were constantly regulated.
All the fine chemicals including B(a)P (purity: ≥96%), DAZ (purity: ≥98%), and solvents were obtained from Sigma-Aldrich (USA).
All animals were randomly divided into five groups with six animals in each group. Group I animals were fed with 0.2 mL of corn oil orally for 16 weeks. Group II animals were orally fed with B(a)P (50 mg/kg b. w. dissolved in maize oil) twice a week for 4 weeks, and then were continued with or without the administration of vehicle for another 12 weeks. Group III mice were administered with DAZ (20 mg/kg b. w. dissolved in corn oil orally) daily for 6 weeks and then B(a)P (as in group II) twice a week for 10 weeks. For the postinitiating experiments, animals in Group IV were administered with B(a)P (as in Group II) for 6 weeks and then with DAZ (as in Group III) for 10 weeks. Histopathological analysis validated the lung carcinoma inductions. DAZ (as in Group III) was administered to Group V animals alone for 16 weeks to verify if DAZ caused any cytotoxicity. The chemopreventive potentials of DAZ in experimental animals were studied using the initiation and postinitiation treatment of DAZ.
Body weight, lung weight, and tumor incidence
The body weight (BW) of the animals was recorded throughout the experimental period, once at the start of the trial and then once a week until the experiment was completed. Following the completion of the experiment, the mice were anesthetized using ketamine/xylazine (90/10 mg/kg). Subsequently, the animals were sacrificed, and their lungs removed, cleaned in saline, and weighed. Tumor incidence (TI) was calculated through manual counting.
Estimation of serum immunoglobins G, A, and M (immunoglobulin G, immunoglobulin A, and immunoglobulin M, respectively)
The serum levels of immunoglobulin G (IgG), immunoglobulin M (IgM), and immunoglobulin A (IgA) in the blood samples of control and treated mice were examined by using the previously described procedures.,
Evaluation of biochemical parameters
The lung homogenate was subjected to the following biochemical analyses: Adenosine deaminase (ADA), aryl hydrocarbon hydroxylase (AHH), gamma glutamyl transferase (GGT), 5′-nucleotidase, and lactate dehydrogenase (LDH).
NADPH-cytochrome c reductase activity was measured using Wharton and Tzagoloff's technique. Omura and Sato's approach was used to calculate the activity of cytochromes P450 and b5.
Phase II enzymes: Assay for detoxification enzymes
The activity of QR was measured using 2,6-dichlorophenol-indophenol as an electron acceptor. The activity of cytosolic fraction of UDP glucuronyltransferase Uridine 5'-diphospho-glucuronosyltransferase (UDP-GT) was evaluated using p-nitrophenol. The glutathione S-transferase (GST) activity was measured using the protocol described by Habig et al.
Evaluation of carcinoembryonic antigen and CK-19 fragment (CYFRA 21-1)
CYFRA 21-1 and carcinoembryonic antigen (CEA) were quantified in the serum samples of normal and treated animals using a chemiluminescent immunoassay on a SIEMENS ADVIA Centaur (Bayer, USA).
Analysis of proinflammatory cytokines
In this study, we measured the level of interleukin (IL) 1 β, IL-6, and tumor necrosis factor alpha (TNF-α) in the cancer tissues. Briefly, tissue homogenate (10%) of the tumor tissue was prepared in phosphate-buffered saline (0.01 M, pH 7.4), and the supernatant was centrifuged for 20 min at 10,000 g. The supernatant was used to quantify the levels of IL1, IL6, and TNF-α (USA).
Lung tissues were removed, weighed, and homogenized in 0.1 M Tris–HCl buffer (pH 7.4). The method described by Johnson and Lardy was used to isolate the mitochondria from the lungs and liver. The total protein content in serum and tissue samples was quantified by the method of Lowry et al.
Evaluation of mitochondrial enzymes
The level of mitochondrial enzymes, namely, isocitrate dehydrogenase (ICDH), α-ketoglutarate dehydrogenase (KDH), succinate dehydrogenase (SDH), and malate dehydrogenase (MDH) (L-malate: NAD oxidoreductase) was measured.
Electron transport chain complex assay
The activities of electron transport chain complexes I II, III, and IV, were measured as described earlier. The method of Lowry et al. was used to estimate the protein content.
Adenosine-5-triphosphate measurements in mice
Adenosine-5-triphosphate (ATP) levels in mice were analyzed using the ATP lite assay kit (Perkin Elmer) immediately after the collection of serum. The assay was conducted as per the manufacturer's instructions. As previously mentioned, (Cicko et al., 2010), the cell lysis phase was avoided to evade the contamination of intracellular ATP.
RNA isolation and reverse transcriptase quantitative polymerase chain reaction assay
For the expression analysis of proliferating cell nuclear antigen (PCNA), NF-κB, CYP1A1, and NRF2, and reverse transcriptase quantitative polymerase chain reaction (RT-qPCR) were used. The RNA was extracted from the tumor tissues with the help of RNA kit, and the assay was conducted based on the manufacturer's protocol. Spectrophotometer was used to determine the total RNA content (Jasco V-700, Japan). We followed the manufacturer's instructions to synthesize cDNA from 2 μg total RNA using the QuantiNova Reverse Transcription Kit (Qiagen, USA). The SYBR green PCR Master Mix (Qiagen, USA) was used to perform qPCR in a Rotor Gene Q 5Plex HRM equipment (Qiagen, USA). The control gene was the actin gene. GenBank was used to create gene-specific primers (NCBI, Bethesda, MD, USA). PCR results were quantified using the ΔCq method.
The lungs of the experimental animals were fixed in 10% formalin and processed for paraffin block production. Hematoxylin and eosin staining was performed on paraffin slices with a thickness of 5 μm. In a microscope (Olympus, Tokyo, Japan), the slides were seen and photographed at ×40 magnification.
Data were analyzed using the SPSS software version 19 (SPSS Inc., IBM Corp, NY, USA). We performed the one-way analysis of variance by Tukey's pos hoc assay to find out the level of significance. The results were displayed as mean ± standard deviation of 6 mice. For significance, P < 0.05 and P < 0.01 were considered statistically significant.
| Results|| |
Impact of daidzein on body weight and lung weight with tumor incidence
[Table 1] shows the data on the general parameters including BW, lung weight, and TI. In comparison with all the groups, Group II mice demonstrated low BW. The mice which were supplemented as preinitiation with DAZ and B(a)P (Group III) had a higher impact on BW related to mice which were supplemented as postinitiation with DAZ and B(a)P (Group IV). Group II mice demonstrated a significantly increased lung weight followed by the Group III and Group IV mice. When compared with the control group, mice in all the treated groups showed an elevation in the lung weight. The TI rate was negligible in both Groups I and V. The TI was significantly high in Group II. There was a significant decrease in the TI in both the DAZ initiation with B(a)P groups.
|Table 1: Effect of daidzein on body weight, lung weight, and tumor incidence in benzo(a)pyrene induced experimental animals|
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Impact of daidzein on Ig levels
[Figure 1]a shows the expression levels of IgG, IgA, and IgM in both control and treated groups. There was a significant decrease in the levels of IgG and IgM in the treated animals when compared to the control animals. The level of IgA was remarkably (P < 0.05) increased in B(a)P-challenged mice. DAZ remarkably increased the levels of IgG and IgM and decreased the levels of IgA in the treated mice. Group V mice did not exhibit any changes in the Ig levels compared with control animals.
|Figure 1: Effect of Daidzein on immunoglobulin levels and tissue maker enzymes in the serum of BaP-induced experimental animals. (a) Immunoglobulin levels (b) tissue marker enzymes. Results are expressed as mean ± standard deviation for six mice in each group. Statistical significance at P < 0.05. * indicates compared with control group (Group I); # indicates compared with Group II|
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Impact of daidzein on tissue marker enzymes and xenobiotic agents
[Figure 1]b shows the results obtained for tissue marker enzymes and xenobiotics (GGT, 5′NT, AHH, LDH) analyzed in the samples obtained from both treated and control animals. All the enzymes were markedly (P < 0.05) augmented in the B(a)P-induced animals in comparison to control group animals. This significant increase might be because of the damage caused to the lungs which increased TI. DAZ supplemented mice showed a reduction in the enzyme activity. Moreover, we did not find variations in the mice that were treated with DAZ alone and control animals.
Impact of daidzein on Phase I and II enzymes in the lungs
[Figure 2]a and [Figure 2]b depicts the expression level of Phase I and II enzymes in the lungs of the control and treated mice. The expression levels of cytochromes P450 and b5, NADPH, and cytochrome P450 reductase were found to be (P < 0.05) increased in the B(a)P-treated animals, and their levels were decreased upon treatment with DAZ. Other enzymes such as UDP glucuronosyltransferase (UDP-GT), GST, and quinone reductase (QR), the expression levels were significantly decreased in the B(a)P-induced mice. DAZ elevated the expressions of these enzymes in the treated animals. Both the control groups and DAZ alone treated groups did not show any deviations in both the phase of the enzymes.
|Figure 2: Effect of Daidzein on phase I and phase II enzymes in the lung of BaP-induced experimental animals. (a) Phase I enzymes (b) Phase II enzymes. Results are expressed as mean ± standard deviation for six mice in each group. Statistical significance at P < 0.05. * indicates compared with the control group (Group I); # indicates compared with Group II|
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Impact of daidzein on tumor markers and cytokines in serum
[Figure 3]a and [Figure 3]b shows the results obtained with respect to tumor markers such as CEA and CYFRA 21-1. The expression of both these markers was significantly (P < 0.05) upregulated in B(a)P-challenged mice. Upon pre- and postadministration with DAZ, the expression levels were downregulated in the serum samples. In addition, the postinitiation group with DAZ showed decreased expression compared with preinitiation group with DAZ. [Figure 3]c shows the data for the estimation of TNF-α, IL-6, and IL-1β in all the groups. Our results show that the expression levels were significantly upregulated in the B(a)P-challenged mice. Furthermore, upon DAZ initiation, the level of all the aforementioned cytokines was decreased. No changes were found in the control and DAZ alone supplemented mice.
|Figure 3: Effect of Daidzein on the levels of tumor markers and cytokines in the serum of BaP induced experimental animals. (a) tumor markers CEA and CYFRA21; (b) tissue maker enzymes adenosine deaminase; (c) pro inflammatory cytokines. The results are expressed as mean ± standard deviation for six mice in each group. Statistical significance at P < 0.05. * indicates compared with control group (Group I); # indicates compared with Group II|
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Effect of daidzein on mitochondrial enzymes in the lungs
[Figure 4]a, [Figure 4]b, [Figure 4]c, [Figure 4]d shows the expression levels of the key mitochondrial enzymes: KDH, ICDH, SDH, and MDH, which were estimated in the lungs of the treated and control animals. According to the results, the mitochondrial enzymes in the B(a)P-challenged animals were found to be significantly downregulated (P < 0.05). According to our results, DAZ increased the activity of these mitochondrial enzymes. We also found that no changes were noted in both control and DAZ alone treated mice.
|Figure 4: Effect of Daidzein on the activity of mitochondrial enzymes in the lung of BaP-induced experimental animals. (a) KDH, (b) ICDH, (c) SDH; (d) MDH Results are expressed as mean ± S.D. for six mice in each group. Statistical significance at P < 0.05. * indicates compared with control group (Gr standard deviation oup I); # indicates compared with Group II.|
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Impact of daidzein on electron transport chain complex and adenosine-5-triphosphate levels in the lungs
[Figure 5]a shows the effect of DAZ in modulating the electron chain transport chain complex, and [Figure 5]b shows the ATP status in the lungs of the animals. According to our results, all four complexes (I, II, III, and IV) and ATP levels were significantly (P < 0.05) downregulated in the B(a)P-treated animals. Pre- and postinitiation of DAZ showed an upregulation of all the complexes and ATP levels. The complexes and ATP levels were not changed in the control and DAZ alone supplemented mice.
|Figure 5: Effect of Daidzein on the activity of electron transport chain complex of the lung in the BaP-induced experimental animals. (a) electron transport chain complex; (b) The levels of ATP in the lung mitochondria. The results are expressed as mean ± standard deviation for six mice in each group. Statistical significance at P < 0.05. * indicates compared with the control group (Group I); # indicates compared with Group II|
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Effect of proliferating cell nuclear antigen, NF-κB, CYP1A1, and NRF2 in lungs
[Figure 7] represents the levels of the key markers including PCNA, NF-κB, CYP1A1, and NRF2 in the lungs of the animals using RT-qPCR analysis. According to the results, the expression levels of markers such as PCNA, NF-κB, and CYP1A1 were significantly upregulated and NRF2 status were significantly downregulated (P < 0.05) in the B(a)P-challenged mice. The expression levels of these markers were modulated in pre- and postinitiation of DAZ in the animals. All the four markers did not show any change in their levels in control and DAZ alone treated animals.
|Figure 7: Effects of Daidzein on proliferating cell nuclear antigen, NF-kB, CYP1AI and NRF-2 mRNA gene expression in benzo(a)pyrene-induced lung cancer. Results are expressed as mean ± standard deviation for six mice in each group. Statistical significance at P < 0.05. * indicates compared with control group (Group I); # indicates compared with Group II|
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Effect of daidzein in histological changes in the lung
[Figure 6] shows the effect of DAZ in histological findings of the lung from control and treated animals. It is observed that control and DAZ alone treated group showed normal histological structures with small nuclei. The B(a)P-induced animals showed lesions exerting proliferations with focal bronchial and hyperplasia in alveolar epithelium. The animals which had pre- and postinitiation of DAZ showed reduced injury in the alveolar region with near-normal arrangements.
|Figure 6: Effect of Daidzein on the histopathological changes of the lung on BaP-induced experimental animals. Control (Group I) and Daidzein alone (Group V) treated group showed normal histomorphological structures without changes. The B(a)P treated animals (Group II) showed lesions (black arrows) exerting abnormal proliferations (blue arrows) with focal bronchial and hyperplasia (yellow arrows) in alveolar epithelium. The animals pre- and postinitiation of Daidzein (Group III and IV) showed diminished injury in the alveolar region with near normal arrangements|
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| Discussion|| |
Most of the lung-related problems are caused due to tobacco smoking. In this study, we aimed to understand the immunomodulatory effect of DAZ in the B(a)P-provoked animals. B(a)P-activated mice develop lung cancer in the treated animals. Previous studies suggest that B(a)P induction leads to loss in BW, and it further enhances the process of carcinogenesis in pulmonary region which is known to be the common sign of tumorigenesis process. One of the major reasons for this weight loss is tumor anorexia cachexia, which is the key player in the inhibition of skeletal muscle and adipose tissue of the animals., Another key factor for this reduction in BW is the higher occurrence of inflammatory nodules. In our study, we found that postinitiation of DAZ helped in maintaining the BW by arresting the TI. We observed that the number of tumors were significantly lower in the DAZ postinitiation group. These results demonstrate that DAZ possess a strong defensive ability and has the potential to arrest the proliferation, tumor growth, and inflammation.
Recent studies suggest that immunomodulation is an efficient alternative strategy for the cancer treatment, which can be achieved by using the bioactive components from plant origin or synthetic chemicals. Our results show that the expression level of IgG and IgM was diminished in cancer, which is a clear indicator of low humoral immune response. These Ig play a key role in neutralizing the toxins, complement activation, and opsonization. The levels of the aforementioned Ig in the serum of patients with cancer are known to be expressed diversely. Studies suggest that factors involved in the hepatic injury may lead to the IgA leakage in serum samples. B(a)P showed immunosuppressive activity in the treated animals and thereby compromising immune system of the animals. After treatment with DAZ, the levels of IgM and IgG were found to be higher and IgA levels were lower than that of control animals. This shows that DAZ possess ameliorative effect.
AHH is known to be cytochrome P-450-dependent tumor-activating enzyme and a key marker for diagnosing the lung cancer. Its activity and expression level are activated by PAHs. A previous study showed that AHH levels were higher in both serum and tissues of B(a)P-provoked animals. LDH is known to be a promising prognostic marker. Previous studies report that there is an increase in the level of LDH in the serum samples obtained from patients with cancer. It plays a key role in the glycolytic pathway, which helps in the production of energy for the tumor to grow. It is also important for tumor survival and growth. Other important tumor-specific markers such as GGT, 5′-NT, and ADA are known to have ability in predicting the process of tumorigenesis. In this study, we found an increase in the cytochromes P450 and b5, NADPH, cytochrome P450 reductase, AHH, LDH, GGT, 5′-NT, and ADA levels in B(a)P-provoked mice, and their levels decreased after DAZ administration.
In general, the level of GST in plasma was elevated due to the hepatic injury and leakage of the cytosolic enzymes that is excreted into the extracellular region.,,, Another important phase II enzyme called UDPGTs is distributed broadly in both hepatic and extrahepatic injury., In this study, we found that these phase II enzymes were restored to their normal levels after DAZ initiation in the treated animals.
There several studies conducted on the inflammatory markers such as CEA, CYFRA 21-1, TNF-α, IL-6, and IL-1β with respect to the development and progression of cancer., In this study, DAZ reduced the levels of these aforementioned markers close to normal, which shows the promising immunomodulatory effects of DAZ.
Enzymes having thiol groups act as receptor sites for the carcinogen to bind, which causes imbalance in the cellular activities. In this study, we found that the enzymes of Trichloroacetic acid (TCA) cycle such as ICDH, MDH, KDH, and SDH were diminished in a B(a)P-challenged mice. The main reason behind this is the changes in the cell morphology and mitochondrial changes happening in the cancer cells. DAZ can balance this situation by increasing the levels of antioxidants.
The electron transport system is known to be the important endogenous source of reactive oxygen species (ROS). ROS has the potential to affect various cellular process which in turn can lead to changes that compromise the integrity and function of normal cells leading to induction of cancer., Our results confirm that DAZ protected the key TCA cycle enzymes and electron transport complexes, thereby the ATP levels were also increased with DAZ initiation.
During the process of tumorigenesis, various transcription factors either get induced or suppressed. NF-κB is the most studied target as it is present in each and every cell performing key functions. Previous studies have shown that B(a)P is functionally activated by CYP1A1 which eventually leads to the formation of DNA adducts, which further increases cancer formation. According to the literature, PCNA is a 33 kDa nuclear protein and is a key player in the proliferation and progression of cancer cells. Our results show that all the key markers were elevated in B(a)P-treated animals which suggest its protective role against cancer conditions. The data from the histopathological analysis of the lung tissue strongly suggest that DAZ initiation reduces the induction of the tumor and ameliorates the factors involved with immuneregulatory action, which might be helpful in the management of lung cancer.
| Conclusion|| |
In conclusion, B(a) P is responsible for the changes in the tissue and serum of the animals with lung cancer. According to our results, it is evident that DAZ initiation in B(a)P-activated lung cancer modulates the antioxidant activities and immune responses of the animals. DAZ might alter the responses of immune cells, either by scavenging or inhibiting the formation of ROS. Overall DAZ possess an immunomodulatory action in B(a)P-induced lung cancer.
Financial support and sponsorship
This work was supported by Integrated TCM and Western Medicine Department, Heilongjiang Provincial Hospital, Harbin, Heilongjiang, 150001, China. This project was supported by Researchers Supporting Project number (RSP-2021/283) King Saud University, Riyadh, Saudi Arabia.
Conflicts of interest
There are no conflicts of interest.
| References|| |
Siegel RL, Miller KD, Jemal A. Cancer statistics, 2020. CA Cancer J Clin 2020;70:7-30.
Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A. Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 2018;68:394-424.
Wu SH, Liu Z. Soy food consumption and lung cancer risk: A meta-analysis using a common measure across studies. Nutr Cancer 2013;65:625-32.
Baena Ruiz R, Salinas Hernández P. Cancer chemoprevention by dietary phytochemicals: Epidemiological evidence. Maturitas 2016;94:13-9.
Yang G, Shu XO, Chow WH, Zhang X, Li HL, Ji BT, et al
. Soy food intake and risk of lung cancer: Evidence from the Shanghai Women's Health Study and a meta-analysis. Am J Epidemiol 2012;176:846-55.
Yang G, Shu XO, Li HL, Chow WH, Wen W, Xiang YB, et al
. Prediagnosis soy food consumption and lung cancer survival in women. J Clin Oncol 2013;31:1548-53.
Wakai K, Ohno Y, Genka K, Ohmine K, Kawamura T, Tamakoshi A, et al
. Risk modification in lung cancer by a dietary intake of preserved foods and soyfoods: Findings from a case-control study in Okinawa, Japan. Lung Cancer 1999;25:147-59.
McCarthy WJ, Meza R, Jeon J, Moolgavkar SH. Chapter 6: Lung cancer in never smokers: Epidemiology and risk prediction models. Risk Anal 2012;32 Suppl 1:S69-84.
Moorthy B, Chu C, Carlin DJ. Polycyclic aromatic hydrocarbons: From metabolism to lung cancer. Toxicol Sci 2015;145:5-15.
Alexandrov K, Rojas M, Satarug S. The critical DNA damage by benzo(a)pyrene in lung tissues of smokers and approaches to preventing its formation. Toxicol Lett 2010;198:63-8.
Perera F. Carcinogenicity of airborne fine particulate benzo(a)pyrene: An appraisal of the evidence and the need for control. Environ Health Perspect 1981;42:163-85.
Hecht SS. Tobacco smoke carcinogens and lung cancer. J Natl Cancer Inst 1999;91:1194-210.
Tan BL, Norhaizan ME, Liew WP, Sulaiman Rahman H. Antioxidant and oxidative stress: A mutual interplay in age-related diseases. Front Pharmacol 2018;9:1162.
Hooper L, Ryder JJ, Kurzer MS, Lampe JW, Messina MJ, Phipps WR, et al
. Effects of soy protein and isoflavones on circulating hormone concentrations in pre- and post-menopausal women: A systematic review and meta-analysis. Hum Reprod Update 2009;15:423-40.
Peng Y, Shi Y, Zhang H, Mine Y, Tsao R. Anti-inflammatory and anti-oxidative activities of daidzein and its sulfonic acid ester derivatives. J Funct Foods 2017;35:635-40.
Cassidy A, Albertazzi P, Lise Nielsen I, Hall W, Williamson G, Tetens I, et al
. Critical review of health effects of soyabean phyto-oestrogens in post-menopausal women. Proc Nutr Soc 2006;65:76-92.
Applegate CC, Rowles JL, Ranard KM, Jeon S, Erdman JW. Soy consumption and the risk of prostate cancer: An updated systematic review and meta-analysis. Nutrients 2018;10:40.
Guo JM, Xiao BX, Liu DH, Grant M, Zhang S, Lai YF, et al
. Biphasic effect of daidzein on cell growth of human colon cancer cells. Food Chem Toxicol 2004;42:1641-6.
Bao C, Namgung H, Lee J, Park HC, Ko J, Moon H, et al
. Daidzein suppresses tumor necrosis factor-α induced migration and invasion by inhibiting hedgehog/Gli1 signaling in human breast cancer cells. J Agric Food Chem 2014;62:3759-67.
Park HJ, Jeon YK, You DH, Nam MJ. Daidzein causes cytochrome c-mediated apoptosis via the Bcl-2 family in human hepatic cancer cells. Food Chem Toxicol 2013;60:542-9.
Jin S, Zhang QY, Kang XM, Wang JX, Zhao WH. Daidzein induces MCF-7 breast cancer cell apoptosis via the mitochondrial pathway. Ann Oncol 2010;21:263-8.
Kasala ER, Bodduluru LN, Barua CC, Madhana RM, Dahiya V, Budhani MK, et al
. Chemopreventive effect of chrysin, a dietary flavone against benzo(a)pyrene induced lung carcinogenesis in Swiss albino mice. Pharmacol Rep 2016;68:310-8.
Tennant B, Baldwin BH, Braun RK, Norcross NL, Sandholm M. Use of the glutaraldehyde coagulation test for detection of hypogammaglobulinemia in neonatal calves. J Am Vet Med Assoc 1979;174:848-53.
Satpathy PK, Dutta NK, Mishra PR, Kai B. Glutaraldehyde coagulation test: Standard curve and its applications to detect gammaglobulin level in kids. Indian Vet J 1996;73:257-60.
Bergmeyer HU. Principles of enzymatic analysis. In: Methods of Enzymatic Analysis. New York: Verlag Chemie; 1984.
Buening MK, Chang RL, Huang MT, Fortner JG, Wood AW, Conney AH. Activation and inhibition of benzo(a)pyrene and aflatoxin B1 metabolism in human liver microsomes by naturally occurring flavonoids. Cancer Res 1981;41:67-72.
Orlowski M, Meister A. Isolation of γ-glutamyl transpeptidase from hog kidney. J Biol Chem 1965;240:338-47.
Luly P, Barnabei O, Tria E. Hormonal control in vitro
of plasma membrane-bound (Na+− K+)-ATPase of rat liver. Biochim Biophys Acta 1972;282:447-52.
King C. The transferases-alanine and aspartate transaminases. In: Van D, editor. Practical Clinical Enzymology. London: Nostrand Company Ltd.; 1965. p. 121-38.
Wharton DC; Tzagoloff A. Cytochrome oxidase from beet heart mitochondria. In: 6th
ed. Methods in enzymology. Vol X. Academic Press, New York; 1967. p. 245-250.
Omura T, Sato R. The carbon monoxide-binding pigment of liver microsomes. I. Evidence for its hemoprotein nature. J Biol Chem 1964;239:2370-8.
Benson AM, Hunkeler MJ, Talalay P. Increase of NAD (P) H: Quinone reductase by dietary antioxidants: Possible role in protection against carcinogenesis and toxicity. Proc Natl Acad Sci 1980;77:5216-20.
Luquita MG, Sánchez Pozzi EJ, Catania VA, Mottino AD. Analysis of p-nitrophenol glucuronidation in hepatic microsomes from lactating rats. Biochem Pharmacol 1994;47:1179-85.
Habig WH, Pabst MJ, Jakoby WB. Glutathione S-transferases. The first enzymatic step in mercapturic acid formation. J Biol Chem 1974;249:7130-9.
Naveenkumar C, Asokkumar S, Raghunandhakumar S, Jagan S, Anandakumar P, Augustine TA, et al
. Potent antitumor and antineoplastic efficacy of baicalein on benzo(a)pyrene-induced experimental pulmonary tumorigenesis. Fundam Clin Pharmacol 2012;26:259-70.
Johnson D, Lardy H. Isolation of liver or kidney mitochondria. Methods Enzymol 1967;10:94-6.
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ. Protein measurement with the Folin phenol reagent. J Biol Chem 1951;193:265-75.
Reed LJ, Mukherjee BB. α-Ketoglutarate dehydrogenase complex from Escherichia coli. Academic Press Inc., New York, Methods Enzymol. 1969;13:55-61.
Slater EC, Borner WD Jr. The effect of fluoride on the succinic oxidase system. Biochem J 1952;52:185-96.
Mehler AH, Kornberg A. The enzymatic mechanism of oxidation-reductions between malate or isocitrate and pyruvate. J Biol Chem 1948;174:961-77.
Birch-Machin MA, Briggs HL, Saborido AA, Bindoff LA, Turnbull DM. An evaluation of the measurement of the activities of complexes I-IV in the respiratory chain of human skeletal muscle mitochondria. Biochem Med Metab Biol 1994;51:35-42.
Krähenbühl S, Talos C, Wiesmann U, Hoppel CL. Development and evaluation of a spectrophotometric assay for complex III in isolated mitochondria, tissues and fibroblasts from rats and humans. Clin Chim Acta 1994;230:177-87.
Capaldi RA, Maruschi MF, Taanman JW. Mammalian cytochrome C oxidase: characterization of enzyme and immunological detection of subunits in tissue extracts and whole cells. Methods Biochem Anal 1995;2:427-34.
Cicko S, Lucattelli M, Müller T, Lommatzsch M, De Cunto G, Cardini S, et al
. Purinergic receptor inhibition prevents the development of smoke-induced lung injury and emphysema. J Immunol 2010;185:688-97.
Pfaffl MW. A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 2001;29:e45.
Bancroft JD, Gamble M. Theory and Practice Histological Techniques. 5th
ed. New York, USA: Churchill Livingstone; 2002.
Underner M, Urban T, Perriot J, de Chazeron I, Meurice JC. Cannabis smoking and lung cancer. Rev Mal Respir 2014;31:488-98.
Velli SK, Sundaram J, Murugan M, Balaraman G, Thiruvengadam D. Protective effect of vanillic acid against benzo(a)pyrene induced lung cancer in Swiss albino mice. J Biochem Mol Toxicol 2019;33:e22382.
Huang L, Zhang P, Duan S, Shao H, Gao M, Zhang Q, et al
. The comparison of two mouse models of inflammation-related lung tumorigenesis induced by benzo(a)pyrene and lipopolysaccharide. Exp Anim 2019;68:301-6.
Inamine T, Schnabl B. Immunoglobulin A and liver diseases. J Gastroenterol 2018;53:691-700.
Chandy KG, Hübscher SG, Elias E, Berg J, Khan M, Burnett D. Dual role of the liver in regulating circulating polymeric IgA in man: Studies on patients with liver disease. Clin Exp Immunol 1983;52:207-18.
Rajendran P, Jayakumar T, Nishigaki I, Ekambaram G, Nishigaki Y, Vetriselvi J, et al
. Immunomodulatory effect of mangiferin in experimental animals with Benzo(a) Pyrene-induced lung carcinogenesis. Int J Biomed Sci 2013;9:68-74.
Rao PS, Kumar S. Polycyclic aromatic hydrocarbons and cytochrome P450 in HIV pathogenesis. Front Microbiol 2015;6:550.
Miao P, Sheng S, Sun X, Liu J, Huang G. Lactate dehydrogenase A in cancer: A promising target for diagnosis and therapy. IUBMB Life 2013;65:904-10.
Asokkumar S, Naveenkumar C, Raghunandhakumar S, Kamaraj S, Anandakumar P, Jagan S, et al
. Antiproliferative and antioxidant potential of beta-ionone against benzo(a)pyrene-induced lung carcinogenesis in Swiss albino mice. Mol Cell Biochem 2012;363:335-45.
Xie H, Hanai J, Ren JG, Kats L, Burgess K, Bhargava P, et al
. Targeting lactate dehydrogenase – A inhibits tumorigenesis and tumor progression in mouse models of lung cancer and impacts tumor-initiating cells. Cell Metab 2014;19:795-809.
Ngo EO, Nutter LM. Status of glutathione and glutathione-metabolizing enzymes in menadione-resistant human cancer cells. Biochem Pharmacol 1994;47:421-4.
Hayes PC, Bouchier IA, Beckett GJ. Glutathione S-transferase in humans in health and disease. Gut 1991;32:813-8.
Tiainen P, Lindgren L, Rosenberg PH. Disturbance of hepatocellular integrity associated with propofol anaesthesia in surgical patients. Acta Anaesthesiol Scand 1995;39:840-4.
Motsch J, Schmidt H, Bach A, Böttiger BW, Böhrer H. Long-term sedation with propofol and green discolouration of the liver. Eur J Anaesthesiol 1994;11:499-502.
Hussey AJ, Howie J, Allan LG, Drummond G, Hayes JD, Beckett GJ. Impaired hepatocellular integrity during general anaesthesia, as assessed by measurement of plasma glutathione S-transferase. Clin Chim Acta 1986;161:19-28.
Gram TE, Okine LK, Gram RA. The metabolism of xenobiotics by certain extrahepatic organs and its relation to toxicity. Annu Rev Pharmacol Toxicol 1986;26:259-91.
Tephly TR, Green MD, Coffman BL, King C, Cheng Z, Rios G. Metabolism of endobiotics and xenobiotics by UDP-glucuronosyltransferase. Adv Pharmacol 1998;42:343-6.
Beauchemin N, Arabzadeh A. Carcinoembryonic antigen-related cell adhesion molecules (CEACAMs) in cancer progression and metastasis. Cancer Metastasis Rev 2013;32:643-71.
Bodduluru LN, Kasala ER, Madhana RM, Barua CC, Hussain MI, Haloi P, et al
. Naringenin ameliorates inflammation and cell proliferation in benzo(a) pyrene induced pulmonary carcinogenesis by modulating CYP1A1, NFκB and PCNA expression. Int Immunopharmacol 2016;30:102-10.
Thirunavukkarasu C, Singh JP, Selvendiran K, Sakthisekaran D. Chemopreventive efficacy of selenium against N-nitrosodiethylamine-induced hepatoma in albino rats. Cell Biochem Funct 2001;19:265-71.
Nohl H, Hegner D. Do mitochondria produce oxygen radicals in vivo? Eur J Biochem 1978;82:563-7.
Collins AR, Duthie SJ, Fillion L, Gedik CM, Vaughan N, Wood SG. Oxidative DNA damage in human cells: The influence of antioxidants and DNA repair. Biochem Soc Trans 1997;25:326-31.
Vuillaume M. Reduced oxygen species, mutation, induction and cancer initiation. Mutat Res 1987;186:43-72.
Chandrashekar N, Selvamani A, Subramanian R, Pandi A, Thiruvengadam D. Baicalein inhibits pulmonary carcinogenesis-associated inflammation and interferes with COX-2, MMP-2 and MMP-9 expressions in-vivo. Toxicol Appl Pharmacol 2012;261:10-21.
Abbass M, Chen Y, Arlt VM, Stürzenbaum SR. Benzo[a]pyrene and Caenorhabditis elegans: Defining the genotoxic potential in an organism lacking the classical CYP1A1 pathway. Arch Toxicol 2021;95:1055-69.
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