Pharmacognosy Magazine

: 2020  |  Volume : 16  |  Issue : 68  |  Page : 181--186

Tumor retardation and immunomodulatory potential of polyherbal formulation HC9 in mouse melanoma model

Snehal Suryavanshi, Kavita Shinde, Prerna Raina, Ruchika Kaul-Ghanekar 
 Cancer Research Laboratory, Interactive Research School for Health Affairs, Bharati Vidyapeeth (Deemed to be University), Pune, Maharashtra, India

Correspondence Address:
Ruchika Kaul-Ghanekar
Cancer Research Lab, Interactive Research School for Health Affairs, Bharati Vidyapeeth (Deemed to be University), Pune-Satara Road, Katraj, Pune - 411 043, Maharashtra


Background: HC9, a polyherbal formulation, is based on Stanyashodhana Kashaya (an Ayurvedic formulation) that is being prescribed by Ayurvedic physicians for the treatment of various disorders of mammary glands. We have recently reported anticancer activity of HC9 in breast cancer cell lines through various molecular mechanisms. Few studies have shown an association between breast cancer and melanoma that has prompted us to find whether HC9 could regulate the melanoma growth as well. Aim of the Study: The aim was to investigate the tumor retardation and immunomodulatory potential of HC9 in mouse melanoma model. Materials and Methods: C57BL/6 mice, with B16F10-induced melanoma tumors, were divided into six groups: tumor control, doxorubicin (2 mg/kg body weight [b.w.]), low dose (100 mg/kg b.w.), intermediate (200 mg/kg b.w.), and high dose (400 mg/kg b.w.) of HC9. No tumor control served as the negative control group. The mice were orally gavaged with HC9 daily for 3 weeks. The urine and blood samples from all the animals were taken before necropsy. The expression of T-helper type 1 (Th1) (interferon-γ and interleukin [IL]-2) and Th2 (IL-4 and IL-10) serum cytokines was evaluated by ELISA assay. Results: HC9 significantly retarded the tumor growth in C57BL/6 mouse melanoma model. The animals did not show any changes in body weight and food consumption throughout the study period. Urine and histopathological analysis revealed no signs of toxicity in HC9-treated animals. HC9 appreciably increased the serum levels of Th1 with a concomitant decrease in Th2 cytokines. Conclusion: HC9 retarded the tumor growth in mouse melanoma model and induced immunomodulation, thereby suggesting the potential of the formulation against melanomas.

How to cite this article:
Suryavanshi S, Shinde K, Raina P, Kaul-Ghanekar R. Tumor retardation and immunomodulatory potential of polyherbal formulation HC9 in mouse melanoma model.Phcog Mag 2020;16:181-186

How to cite this URL:
Suryavanshi S, Shinde K, Raina P, Kaul-Ghanekar R. Tumor retardation and immunomodulatory potential of polyherbal formulation HC9 in mouse melanoma model. Phcog Mag [serial online] 2020 [cited 2022 Dec 6 ];16:181-186
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Full Text


  • HC9 retarded tumor growth in C57BL/6 mouse melanoma model
  • HC9 increased the serum level of Th1 cytokines and decreased Th2 cytokines
  • There was no difference in the body weight and food consumption of animals treated with HC9
  • Urine and histopathological analysis revealed no signs of toxicity in HC9-treated animals.


Abbreviations used: HC9: Herbal composition comprised of 9 medicinal plants; TC: Tumor control; Dox: Doxorubicin; NTC: No Tumor Control; IFN-γ: Interferon-gamma; IL: Interleukin; NF-κB: Nuclear factor kappa-light-chain-enhancer of activated B-cells; COX-2: Cyclooxygenase-2; MMP: Matrix metalloproteinase; HIF1-α: Hypoxia-inducible factor 1-alpha; VEGF: Vascular endothelial growth factor; SMAR1: Scaffold/Matrix-associated region-1; CDP/Cux: CCAAT displacement protein/cut homeobox.


Epidemiological studies have suggested a relationship between breast cancer and cutaneous melanoma.[1] Interestingly, carriers of genetic mutations in BRCA2 gene have an increased risk of developing melanoma, whereas those having mutations in CDKN2A (melanoma susceptibility) gene have a higher propensity to develop breast cancer. It was reported that in young breast cancer patients, there was a 46% elevated risk of developing a second melanoma. Thus, there seems to be an overlap between the pathways underlying the two types of cancers.

Herbal drugs either as single herbs or polyherbal formulations have been shown to exhibit anticancer activity against various cancers. These drugs exhibit various biological activities and are easily accessible, cost-effective, and safe.[2] However, due to lack of extensive scientific and clinical evidences, herbal drugs have not yet found clinical application.[3] Thus, it becomes essential to ascertain the safety and therapeutic efficacy of herbal medicines, particularly, polyherbal formulations at preclinical level.

HC9 polyherbal formulation has been developed based on Stanya Shodhana Kashaya (SSK). The latter has been prescribed by Ayurvedic practitioners mainly for the breast milk detoxification and lactation-related disorders (Charak Samhita, Chapter 22).[4] The original SSK formulation is composed of ten medicinal plants: Picrorhiza kurroa Royle ex Benth., Cyperus rotundus L., Zingiber officinale Roscoe, Cedrus deodara (Roxb. ex D. Don) G. Don, Tinospora cordifolia (Willd.) Miers, Holarrhena antidysenterica (Roth) Wall. ex A.DC., Swertia chirata Buch.-Ham. ex Wall., Cyclea peltata (Lam.) and Hemidesmus indicus (L.) R. Br. ex Schult, and Marsdenia tenacissima (Roxb.) Moon. In HC9 formulation, only nine plants have been included because of the non-availability of M. tenacissima, which is found only in the Himalayan region. HC9 was previously standardized and found to contain marker compounds such as picroside-I, nootkatone, 6-gingerol, matairesinol, swertiamarin, berberine, connesine, and 2-hydroxy-4-methoxybenzaldehyde.[5] HC9 exhibited antioxidant activity and significantly reduced the viability of MCF-7 and MDA-MB-231.[5] Acute and subacute toxicity studies in Swiss albino have revealed the safety of HC9 up to 2000 mg/kg body weight (b.w.) of mice with no adverse effects.[6] Recently, we have reported that HC-induced cell cycle arrest reduced migration and expression of angiogenic markers in breast cancer cells.[7] It also regulated the expression of chromatin and inflammatory marker proteins. In the present study, we have evaluated the anticancer activity and immunomodulatory potential of HC9 in a mouse melanoma model.

 Materials and Methods

Plant material

The nine different plant materials of HC9, such as P. kurroa (R-120), C. rotundus (R-121), Z. officinale (R-122), C. deodara (S/B-096), T. cordifolia (S/B-097), H. antidysenterica (S-119), S. chirata (WP-078), Cissampelos pareira (Medicinal Plant Conservation Centre [MPCC] 290), and H. indicus (MPCC 2354), were purchased from Shri Shailya Medi Pharms, Solapur, Maharashtra, India. They were botanically authenticated at the Department of Botany, Agharkar Research Institute and Herbaria of MPCC, Pune.[7],[8],[9]

Preparation of ethanolic extract

HC9 formulation was made by mixing equal parts (1:1 ratio) of each plant material and extracted in ethanol by Soxhlet apparatus as described previously.[7],[8],[9],[10]

Experimental animals

The study was sanctioned by the Institutional Animal Ethics Committee (CPCSE Reg. No. 258/CPCSE), Bharati Vidyapeeth University, Pune. Female C57BL/6 mice, 6–8 weeks old with an average weight of 18-22g, were procured from the National Institute of Nutrition (Hyderabad, India). The mice were divided into different groups and housed in polypropylene cages at 21°C ± 3°C with relative humidity of 30%–70% and 12:12 h light/dark rhythm. The mice were acclimatized to laboratory conditions and fed with commercial food pellets (Nutrivet, Pune) and water ad libitum.

Tumor induction and its assessment

The tumors were raised in mice by subcutaneous injection of 0.2 ml of B16F10 cells (5 × 105 cells/animal) into the right flank region. Tumors were palpable at the 8th day after the injection of cells after which the animals were grouped with n = 4/group. Groups I and II were no tumor control (NTC) and tumor control (TC), respectively, which received distilled water. Group III was positive control (PC) which received doxorubicin (Dox) (intravenous [i.v.], 2 mg/kg b.w.) on the 1st, 5th, and 9th days after development of tumors. Groups IV, V, and VI were orally gavaged with low (100 mg/kg b.w.), intermediate (200 mg/kg b.w.), and high (400 mg/kg b.w.) doses of HC9, respectively, daily for 2 weeks. Food consumption, body weights, and tumor sizes were recorded after every 3 days. Tumor volume (mm 3) was calculated as: 0.5 × shortest diameter 2 × largest diameter. Percentage of tumor growth inhibition was calculated as: (Average tumor volume of control group − Average tumor volume of test group)/(Average tumor volume of control group) ×100. The urine and blood samples were collected before necropsy of animals. After necropsy, different organs were collected for histopathological analysis.

Determination of serum cytokine levels

The serum cytokines (interleukin [IL]-2, IL-4, IL-10, and interferon [IFN]-γ) levels were determined using mouse T-helper type 1/T-helper type 2 (Th1/Th2) ELISA-Ready-Set-Go (eBioscience, San Diego, CA, USA) kit after following the manufacturer's instructions.[11] Readings were taken at 450 nm with FLUOstar Omega microplate reader (CA, USA).

Statistical analysis

The data were analyzed using GraphPad Prism 5 software (GraphPad Software, Inc., San Diego, CA, USA). The experiment was done once, and values with *P < 0.05, **P < 0.01, and ***P < 0.001 were considered to be statistically significant.


HC9 retarded tumor growth

Oral administration of HC9 significantly reduced the tumor growth in the mice. After comparing with the TC group, mice treated with Dox, HC9 100, 200, and 400 showed significant reduction in the tumor volume by ~65.3 ± 28.3% (P < 0.0001), 53.2% ± 17.45% (P < 0.05), 47.3% ± 23.7% (P < 0.05), and 76.5% ± 12.7% (P < 0.0001), respectively [Figure 1].{Figure 1}

HC9 was safe for the animals

No significant change was observed in the body weight [Table 1] and food intake [Table 2] of animals treated with either Dox or different doses (100, 200, and 400 mg/kg b.w.) of HC9 compared to NTC or TC. No significant difference (P > 0.05) in the relative organ weights of the treated mice was observed compared to either NTC or TC mice. The urine analysis revealed no signs of toxicity in the HC9-treated mice [Table 3]a, [Table 3]b, [Table 3]c, [Table 3]d, [Table 3]e, [Table 3]f.{Table 1}{Table 2}{Table 3}

HC9 modulated T-helper type 1/T-helper type 2 cytokine levels

It was observed that compared to the TC group, at 400 mg/kg dose, HC9 increased IFN-γ and IL-2 levels by ~1.7 (P > 0.05; NS) and ~2.7 (P < 0.05) folds, respectively [Figure 2]a. At 400 mg/kg dose, HC9 decreased IL-10 and IL-4 levels by ~1.2 (P > 0.05; NS) and ~4.4 (P < 0.0001) folds, respectively, compared to the TC group [Figure 2]b.{Figure 2}


The current study reported the potential of HC9, a polyherbal formulation on tumor retardation and regulation of immunomodulatory cytokines in a mouse melanoma model. We have recently shown that HC9 demonstrated a significant anticancer activity against breast cancer cells through various mechanisms. Here, we have evaluated the effect of HC9 against melanoma. Various studies have reported an increased risk for skin cancer in patients having an earlier episode of breast cancer.[8] Female breast cancer survivors <45 years of age showed a 38% higher risk of developing melanoma as a second cancer than the general population.[9] Thus, newer drugs need to be explored for targeting not only breast cancer but also for preventing the recurrence of secondary cancers such as melanoma.

Interestingly, HC9 retarded tumor growth in mouse melanoma model. HC9 did not have any potential adverse effect on either mouse survival or organ pathology. Moreover, the tumor retardation potential of HC9 was comparable to that of the PC Dox. Our recent work has shown that HC9 modulated the expression of cell cycle, inflammation, and chromatin regulatory proteins. It decreased migration and invasion of breast cancer cells through modulation of matrix metalloproteinase (MMP)-2, MMP-9, hypoxia-inducible factor 1-alpha, and vascular endothelial growth factor expression.[7] We have reported earlier that HC9 contained picroside I, 6-gingerol, matairesinol, connesine, swertiamarin, berberine, and 2-hydroxy-4-methoxy-benzaldehyde found in P. kurroa, C. rotundus, Z. officinale, C. deodara, T. cordifolia, H. antidysenterica, S. chirata, C. pareira, and H. indicus, respectively.[5] Recently, these bioactives have been shown to decrease the viability of breast cancer cells (communicated). Moreover, compared to other bioactives, MA exhibited lower IC50 values. Thus, the anticancer activity of HC9 could be attributed to its component bioactives. Further, the individual plant materials of the formulation have been reported to exhibit anticancer activity. For example, P. kurroa,[10]C. rotundus,[12]Z. officinale,[13]T. cordifolia,[14]H. indicus,[15] and C. deodara[16] have been reported to retard the growth of Ehrlich ascites carcinoma-induced tumors in mice. H. antidysenterica has exhibited anticancer activity against human OVCAR-5 (ovary), HT-29 (colon), SK-N-MC (neuroblastoma), HEP-2 (liver), COLO-205 (colon), NIH-OVCAR-3 (ovary), and A-549 (lung) cell lines.[17]S. chirata has exhibited anticancer activity in DMBA-induced mouse skin carcinogenesis model.[18]C. peltata showed anticancer activity against human breast carcinoma cells.[19] All these studies suggested that the bioactives present in HC9 formulation could be regulating the melanoma growth through modulation of various signal transduction mechanisms.

Various herbal formulations have been shown to exhibit anticancer activity against melanoma models. For example, a herbal composition having Sophorae Flos and Lonicerae Japonicae Flos has been reported to inhibit melanoma growth in vivo.[20] Other plants such as Artemisia annua,[21]Hedyotis diffusa,[22] and Rosmarinus officinalis[23] have also been reported to inhibit the proliferation, invasion, migration, and angiogenesis of melanoma cells in vitro. Curcuma rhizome exhibited antiproliferative and proapoptotic activities in B164A5 melanoma cells.[24]Ganoderma lucidum exhibited antimelanoma activity in vitro and in vivo, by inducing oxidative stress, apoptosis, and inhibition of cell proliferation.[25]Coptidis rhizoma water extracts or its major active chemical component, berberine, showed activity against human melanoma cells.[26] The coumarin fraction of Cachrys pungens Jan, was shown to be useful in the treatment of melanoma and nonmelanoma skin cancers.[27] Aerial components of Ficus carica L. cultivar Dottato (F. carica) exhibited antioxidant and phototoxic activities in human melanoma cells.[28]

HC9 modulated Th1 and Th2 cytokine response in the mice. These cytokines reflect immune response in various human diseases, including cancer.[29] Several studies have shown that there is a shift from Th1 to Th2 cytokines in cancer patients undergoing chemo- and radiotherapies that usually lead to immunosuppression.[30],[31] Recently, plant extracts have been reported to modulate Th1 and Th2 expression in cancer.[32],[33] HC9 upregulated IFN-γ and IL-2 (Th1) cytokines and reduced IL-10 and IL-4 (Th2) cytokine levels in mice. Resveratrol has been shown to improve the efficacy of IL-2 immunotherapy against melanoma model in vivo.[34] IFN-γ is an important lymphokine that can activate the immune cells (natural killer cells, cytotoxic T-lymphocytes, and tumoricidal macrophages), which preferentially attack breast cancer,[35] melanoma,[36] neuroblastoma,[37] and methylcholanthrene-induced tumors [38] by activation of CD8+ cytotoxic T-cells. IL-2 is a tumor suppressor/cytokine with pleiotropic effects on the immune system including monocytes and activated T-cells. IL-2 has been reported to suppress the growth of melanoma cells through activation of IL-24.[39] On the other hand, IL-4 and IL-10 contribute to tumor aggressiveness, induce immunosuppression, and help in avoiding tumor immune surveillance.[40],[41] Some plants have been shown to inhibit the growth of melanoma cells through modulation of immune response. T. cordifolia has been shown to activate tumor-associated macrophages and induce antitumor activity in the spontaneous T-cell lymphoma.[42]Astragalus membranaceus has been shown to exhibit immune regulation by enhancing NK cell activity, inducing lymphocyte-mediated killing of tumors, and stimulating macrophage and B-cell activities.[43] Similarly, the aqueous extract of Daphne gnidium was reported to exhibit antitumor and immunomodulatory activities in mouse melanoma model.[44]


The current report indicated the antitumor and immunomodulatory potential of HC9 in melanoma model although it has been earlier reported to exhibit antibreast cancer activity. Herbal drugs that would target multiple cancers could be a boon to the patients as such drugs could prevent recurrence with second cancers.


We thank IRSHA, Bharati Vidyapeeth (Deemed to be University), for providing the infrastructure for completing this work.

Financial support and sponsorship

We thank IRSHA, BVDU, Pune for financially supporting this work. Snehal Suryavanshi, ICMR-SRF, would like to thank Indian Council of Medical Research (ICMR) for financial assistance.

Conflicts of interest

There are no conflicts of interest.


1Goggins W, Gao W, Tsao H. Association between female breast cancer and cutaneous melanoma. Int J Cancer 2004;111:792-4.
2Metri K, Bhargav H, Chowdhury P, Koka PS. Ayurveda for chemo-radiotherapy induced side effects in cancer patients. J Stem Cells 2013;8:115-29.
3Zhang AL, Xue CC, Fong HH. Integration of herbal medicine into evidence-based clinical practice. In: Herbal Medicine: Biomolecular and Clinical Aspects. 2nd ed. United Kingdom: CRC Press/Taylor and Francis; 2011.
4Shastri SS. Charak Samhita with elaborated Vidhyotini Hindi commentary Sutrasthan Langhanbruhmniya Adhyaya. Bharty academy: Varanasi Chukhambha; 2014. p. 429.
5Suryavanshi S, Zanwar A, Hegde M, Kaul-Ghanekar R. Standardization of a polyherbal formulation (HC9) and comparative analysis of its cytotoxic activity with the individual herbs present in the composition in breast cancer cell lines. Pharmacog J 2014;6:87-95.
6Suryavanshi SA, Kadam SK, Raina P, Nimbargi R, Pandit VA. Evaluation of acute and sub-acute toxicity of a standardized polyherbal formulation (Hc9): An in vivo study. Int J Pharm Pharm Sci 2015;7:110-7.
7Suryavanshi S, Choudhari A, Raina P, Kaul-Ghanekar R. A polyherbal formulation, HC9 regulated cell growth and expression of cell cycle and chromatin modulatory proteins in breast cancer cell lines. J Ethnopharmacol 2019;242:1-10.
8Drueppel D, Schultheis B, Solass W, Ergonenc H, Tempfer CB. Primary malignant melanoma of the breast: Case report and review of the literature. Anticancer Res 2015;35:1709-13.
9Yang GB, Barnholtz-Sloan JS, Chen Y, Bordeaux JS. Risk and survival of cutaneous melanoma diagnosed subsequent to a previous cancer. Arch Dermatol 2011;147:1395-402.
10Soni D, Grover A. Picrosides from Picrorhiza kurroa as potential anti-carcinogenic agents. Biomed Pharmacother 2019;109:1680-7.
11Deshpande R, Raina P, Shinde K, Mansara P, Karandikar M, Kaul-Ghanekar R. Flax seed oil reduced tumor growth, modulated immune responses and decreased HPV E6 and E7 oncoprotein expression in a murine model of ectopic cervical cancer. Prostaglandins Other Lipid Mediat 2019;143:1-10.
12Nidugala H, Avadhani R, Prabhu A, Basavaiah R. In vitro cytotoxic activity of rhizome extracts of Cyperus rotundus (L.) against colon carcinoma and Ehrlich ascites carcinoma. J Appl Pharm Sci 2016;6:172-5.
13John J. Cytotoxic Antitumor Antioxidant and Phytochemical Assays in some Species of Zingiber boehm. Department of Botany: University of Calicut; 2014.
14Sarkar R, Mandal N. In vitro cytotoxic effect of hydroalcoholic extracts of medicinal plants on Ehrlich's Ascites Carcinoma (EAC). Int J Phytomed 2011;3:370-80.
15Zarei M, Javarappa KK. Anticarcinogenic and cytotoxic potential of Hemidesmus indicus root extract against Ehrlich Ascites tumor. Der Pharmacia Lettre 2012;4:906-10.
16Chaudhary AK, Ahmad S, Mazumder A. Cedrus deodara (Roxb.) Loud: A review on its ethnobotany, phytochemical and pharmacological profile. Pharmacog J 2011;3:12-7.
17Sharma V, Hussain S, Bakshi M, Bhat N, Saxena AK. In vitro cytotoxic activity of leaves extracts of Holarrhena antidysenterica against some human cancer cell lines. Indian J Biochem Biophys 2014;51:46-51.
18Saha P, Mandal S, Das A, Das PC, Das S. Evaluation of the anticarcinogenic activity of Swertia chirata Buch. Ham, an Indian medicinal plant, on DMBA-induced mouse skin carcinogenesis model. Phytother Res 2004;18:373-8.
19Bhagya N, Chandrashekar KR, Prabhu A, Rekha PD. Tetrandrine isolated from Cyclea peltata induces cytotoxicity and apoptosis through ROS and caspase pathways in breast and pancreatic cancer cells. In vitro Cell Dev Biol Anim 2019;55:331-40.
20Li T, Fu X, Tse AK, Guo H, Lee KW, Liu B, et al. Inhibiting STAT3 signaling is involved in the anti-melanoma effects of a herbal formula comprising Sophorae Flos and Lonicerae japonicae flos. Sci Rep 2017;7:3097.
21AlQathama A, Prieto JM. Natural products with therapeutic potential in melanoma metastasis. Nat Prod Rep 2015;32:1170-82.
22Ling B, Michel D, Sakharkar MK, Yang J. Evaluating the cytotoxic effects of the water extracts of four anticancer herbs against human malignant melanoma cells. Drug Des Devel Ther 2016;10:3563-72.
23Russo A, Lombardo L, Troncoso N, Garbarino J, Cardile V. Rosmarinus officinalis extract inhibits human melanoma cell growth. Nat Prod Commun 2009;4:1707-10.
24Danciu C, Vlaia L, Fetea F, Hancianu M, Coricovac DE, Ciurlea SA, et al. Evaluation of phenolic profile, antioxidant and anticancer potential of two main representants of Zingiberaceae family against B164A5 murine melanoma cells. Biol Res 2015;48:1.
25Harhaji Trajković LM, Mijatović SA, Maksimović-Ivanić DD, Stojanović ID, Momcilović MB, Tufegdzić SJ, et al. Anticancer properties of Ganoderma lucidum methanol extracts in vitro and in vivo. Nutr Cancer 2009;61:696-707.
26Park SY, Song H, Sung MK, Kang YH, Lee KW, Park JH. Carnosic acid inhibits the epithelial-mesenchymal transition in B16F10 melanoma cells: A possible mechanism for the inhibition of cell migration. Int J Mol Sci 2014;15:12698-713.
27Menichini G, Alfano C, Provenzano E, Marrelli M, Statti GA, Menichini F, et al. Cachrys pungens Jan inhibits human melanoma cell proliferation through photo-induced cytotoxic activity. Cell Prolif 2012;45:39-47.
28Conforti F, Menichini G, Zanfini L, Tundis R, Statti GA, Provenzano E, et al. Evaluation of phototoxic potential of aerial components of the fig tree against human melanoma. Cell Prolif 2012;45:279-85.
29Annunziato F, Cosmi L, Liotta F, Maggi E, Romagnani S. Human Th1 dichotomy: Origin, phenotype and biologic activities. Immunology 2014;144:343-51.
30van Meir H, Nout RA, Welters MJ, Loof NM, de Kam ML, van Ham JJ, et al. Impact of (chemo) radiotherapy on immune cell composition and function in cervical cancer patients. Oncoimmunology 2017;6:e1267095.
31Lippitz BE. Cytokine patterns in patients with cancer: A systematic review. Lancet Oncol 2013;14:e218-28.
32Kartini, Piyaviriyakul S, Thongpraditchote S, Siripong P, Vallisuta O. Effects of Plantago major extracts and its chemical compounds on proliferation of cancer cells and cytokines production of lipopolysaccharide-activated THP-1 macrophages. Pharmacogn Mag 2017;13:393-9.
33Burns JJ, Zhao L, Taylor EW, Spelman K. The influence of traditional herbal formulas on cytokine activity. Toxicology 2010;278:140-59.
34Guan H, Singh NP, Singh UP, Nagarkatti PS, Nagarkatti M. Resveratrol prevents endothelial cells injury in high-dose interleukin-2 therapy against melanoma. PLoS One 2012;7:e35650.
35Nagai Y, Tsuchiya H, Ji MQ, Zhang H, Greene MI. Synergistic effect of IFN-γ on breast cancer targeted therapy. J Immunol 2018;198:141-14.
36Zippelius A, Batard P, Rubio-Godoy V, Bioley G, Liénard D, Lejeune F, et al. Effector function of human tumor-specific CD8 T cells in melanoma lesions: A state of local functional tolerance. Cancer Res 2004;64:2865-73.
37Reid GS, Shan X, Coughlin CM, Lassoued W, Pawel BR, Wexler LH, et al. Interferon-gamma-dependent infiltration of human T cells into neuroblastoma tumors in vivo. Clin Cancer Res 2009;15:6602-8.
38Nishikawa H, Kato T, Tawara I, Ikeda H, Kuribayashi K, Allen PM, et al. IFN-gamma controls the generation/activation of CD4+ CD25+ regulatory T cells in antitumor immune response. J Immunol 2005;175:4433-40.
39Jen EY, Poindexter NJ, Farnsworth ES, Grimm EA. IL-2 regulates the expression of the tumor suppressor IL-24 in melanoma cells. Melanoma Res 2012;22:19-29.
40Hassuneh MR, Nagarkatti M, Nagarkatti PS. Role of interleukin-10 in the regulation of tumorigenicity of a T cell lymphoma. Leuk Lymphoma 2013;54:827-34.
41Rawal S, Park HJ, Chu F, Zhang M, Nattamai D, Kannan SC, et al. Role of IL-4 in inducing immunosuppressive tumor microenvironment in follicular lymphoma. Blood 2011;118:-771.
42Singh N, Singh SM, Shrivastava P. Immunomodulatory and antitumor actions of medicinal plant Tinospora cordifolia are mediated through activation of tumor-associated macrophages. Immunotoxicol 2004;26:145-62.
43Cho WC, Leung KN. In vitro and in vivo anti-tumor effects of Astragalus membranaceus. Cancer Lett 2007;252:43-54.
44Chaabane F, Mustapha N, Mokdad-Bzeouich I, Sassi A, Kilani-Jaziri S, Dijoux Franca MG, et al. In vitro and in vivo anti-melanoma effects of Daphne gnidium aqueous extract via activation of the immune system. Tumour Biol 2016;37:6511-7.