Antiproliferative and apoptotic effect of herbal formulation containing black and green tea extract against HCT-116 human colon cancer cells

Main Article Content

S.Jeswin Immanuel
Parameswari R P
Lakshmi Thangavelu


black tea, green tea, polyphenols, colon cancer, antiproliferative, apoptosis


Background: Due to their safety, greater tolerance, and economics, phytochemical-based dietary interventions have drawn attention as a preventive and therapeutic method against most cancers. The purpose of this study was to investigate the antiproliferative and apoptotic effect of black and green tea formulation.
Materials and Methods: Aqueous extracts of black and tea were prepared. The formulation was prepared by mixing 1:1 part of individual aqueous extracts of black and tea. The prepared formulation was evaluated for in vitro cytotoxicity using HCT-116 colon cancer cells. Morphological changes were observed and the apoptotic effect of the formulation was also assessed.
Results: The results showed that the formulation was able to inhibit the growth of colon cancer cells effectively at all the tested concentrations between the range of 5-50μg/ml. A dose-dependent decrease in the viability of cells was observed. IC50 was calculated by linear regression analysis. Based on the IC50, efficacy concentration of 45μg/ml was chosen for morphology and dual staining assays. Morphological changes were observed after treating the cells at a single concentration of 45μg/ml. The treated cells demonstrated loosely adhered boundaries with spherical morphology indicating the onset of the cell death following the black and green tea formulation treatment. To further confirm the antiproliferative effect, the formulation treated cells were stained with ethidium bromide/acridine orange and assessed for apoptotic effect. The images showed greenish yellow fluorescent cells in addition to few green fluorescent cells when compared to untreated cells that appeared only green in fluorescence.
Conclusion: Hence from the results of the present study it may be concluded that the black and green tea formulation were found to show significant antiproliferative and apoptotic effect in HCT 116 colon cancer cells.

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1. Siegel RL, Wagle NS, Cercek A, Smith RA, Jemal A. Colorectal cancer statistics, 2023. CA Cancer J Clin. 2023 Mar 1. doi: 10.3322/caac.21772.
2. Islami F, Goding Sauer A, Miller KD, et al. Proportion and number of cancer cases and deaths attributable to potentially modifiable risk factors in the United States. CA Cancer J Clin. 2018;68(1):31-54.
3. Namrata, D., Lakshmi, T., Rushabh, D. and Ezhilarasan, D., 2022. Role of stress management in oral malignant and premalignant conditions. Oral Oncology, 127, pp.105813-105813.
4. Ramakrishnan, D.S., Gouthaman, S.S. and Muthusekhar, M.R., 2022. Ameloblastic fibrosarcoma transformation from ameloblastic fibroma. National Journal of Maxillofacial Surgery, 13(Suppl 1), pp.S145-S149.
5. Jasperson, K.W., Tuohy, T.M., Neklason, D.W. and Burt, R.W., 2010. Hereditary and familial colon cancer. Gastroenterology, 138(6), pp.2044-2058.
6. Amersi, F., Agustin, M. and Ko, C.Y., 2005. Colorectal cancer: epidemiology, risk factors, and health services. Clinics in colon and rectal surgery, 18(03), pp.133-140.
7. Singh, S., Ramani, P., Jayakumar, N.D., Pannu, S.J., Sharma, R.K. and Gill, S.S., 2022. Role of Pro-inflammatory and Anti-inflammatory Cytokines in Pathophysiology of Psoriasis. Current Oral Health Reports, 9(4), pp.132-145.
8. Sahoo, N.K., Sahu, M., Pullaiah, C.P. and Muralikrishna, K.S., 2020. In vitro anticancer activity of Eclipta alba whole plant extract on colon cancer cell HCT-116. BMC Complementary Medicine and Therapies, 20(1), pp.1-8.
9. Kim, H.S., Quon, M.J. and Kim, J.A., 2014. New insights into the mechanisms of polyphenols beyond antioxidant properties; lessons from the green tea polyphenol, epigallocatechin 3-gallate. Redox biology, 2, pp.187-195.
10. Joshua D Lambert, Does tea prevent cancer? Evidence from laboratory and human intervention studies, The American Journal of Clinical Nutrition, Volume 98, Issue 6, December 2013, Pages 1667S–1675S.
11. Wang, L.F., Lee, J.Y., Chung, J.O., Baik, J.H., So, S. and Park, S.K., 2008. Discrimination of teas with different degrees of fermentation by SPME–GC analysis of the characteristic volatile flavour compounds. Food chemistry, 109(1), pp.196-206.
12. Beltz LA, Bayer DK, Moss AL, Simet IM. Mechanisms of cancer prevention by green and black tea polyphenols. Anticancer Agents Med Chem. 2006 Sep;6(5):389-406.
13. Pan, M.H., Lai, C.S., Wang, H., Lo, C.Y., Ho, C.T. and Li, S., 2013. Black tea in chemo-prevention of cancer and other human diseases. Food Science and Human Wellness, 2(1), pp.12-21.
14. Imran, A., Butt, M.S., Xiao, H., Imran, M., Rauf, A., Mubarak, M.S. and Ramadan, M.F., 2019. Inhibitory effect of black tea (Camellia sinensis) theaflavins and thearubigins against HCT 116 colon cancer cells and HT 460 lung cancer cells. Journal of food biochemistry, 43(5), p.e12822.
15. Min, K.J. and Kwon, T.K., 2014. Anticancer effects and molecular mechanisms of epigallocatechin-3-gallate. Integrative medicine research, 3(1), pp.16-24.
16. Gurumurthy, Kaviyaselvi, S. Balaji Ganesh, S. Jayalakshmi, and S. Sasidharan. "Effects Of Black Tea and Coffee on the Colour Stability of Glass Ionomer Cement - An in vitro study." International Journal of Dentistry and Oral Science (2021): 4642-4647.
17. Rady, I., Mohamed, H., Rady, M., Siddiqui, I.A. and Mukhtar, H., 2018. Cancer preventive and therapeutic effects of EGCG, the major polyphenol in green tea. Egyptian Journal of Basic and Applied Sciences, 5(1), pp.1-23.
18. Lu, J. B., Ho, C. T., Ghai, G., & Chen, K. Y. (2000). Differential effects of theaflavin monogallates on cell growth, apoptosis and Cox‐2 gene expression in cancerous versus normal cells. Cancer Research, 60, 6465–6471.
19. Suganya, M., Gnanamangai, B.M., Ravindran, B., Chang, S.W., Selvaraj, A., Govindasamy, C., Elsadek, M.F. and Ponmurugan, P., 2019. Antitumor effect of proanthocyanidin induced apoptosis in human colorectal cancer (HT-29) cells and its molecular docking studies. BMC chemistry, 13(1), pp.1-14.
20. Long, J., Guan, P., Hu, X., Yang, L., He, L., Lin, Q., Luo, F., Li, J., He, X., Du, Z. and Li, T., 2021. Natural polyphenols as targeted modulators in colon cancer: molecular mechanisms and applications. Frontiers in Immunology, 12, p.635484.
21. Ezhilarasan, D., Apoorva, V.S. and Ashok Vardhan, N., 2019. Syzygium cumini extract induced reactive oxygen species‐mediated apoptosis in human oral squamous carcinoma cells. Journal of Oral Pathology & Medicine, 48(2), pp.115-121.
22. Saldanha, S.N., Kala, R. and Tollefsbol, T.O., 2014. Molecular mechanisms for inhibition of colon cancer cells by combined epigenetic-modulating epigallocatechin gallate and sodium butyrate. Experimental cell research, 324(1), pp.40-53.
23. Kumar, S.A. and Ramachandra Prabhakar, N., 2021. Raj Vikram, SP Saravana Dinesh, S Rajeshkumar. Antioxidant Activity of Silymarin/Hydroxyapatite/Chitosan Nano Composites-An In Vitro Study. Int J Dentistry Oral Sci, 8(1), pp.1402-1404.
24. Nasim, I., Jabin, Z., Kumar, S.R. and Vishnupriya, V., 2022. Green synthesis of calcium hydroxide-coated silver nanoparticles using Andrographis paniculata and Ocimum sanctum Linn. leaf extracts: An antimicrobial and cytotoxic activity. Journal of Conservative Dentistry, 25(4), p.369.
25. Kumar S, L., Naik, Z., Panwar, A., Keluskar, V. and Kumar, R.S., 2022. Comparative evaluation of the efficacy of Nigella sativa (75% v/v) cream and clobetasol propionate (0.05% w/w) gel in oral lichen planus—a double-blinded randomized control trial. Oral and Maxillofacial Surgery, pp.1-10.
26. Panji, M., Behmard, V., Zare, Z., Malekpour, M., Nejadbiglari, H., Yavari, S., Safaeian, A., Bakhshi, A., Abazari, O., Abbasi, M. and Khanicheragh, P., 2021. Synergistic effects of green tea extract and paclitaxel in the induction of mitochondrial apoptosis in ovarian cancer cell lines. Gene, 787, p.145638.
27. Trisha, A.T., Shakil, M.H., Talukdar, S., Rovina, K., Huda, N. and Zzaman, W., 2022. Tea Polyphenols and Their Preventive Measures against Cancer: Current Trends and Directions. Foods, 11(21), p.3349.
28. Marimuthu, M., Andiappan, M., Wahab, A., Muthusekhar, M.R., Balakrishnan, A. and Shanmugam, S., 2018. Canonical Wnt pathway gene expression and their clinical correlation in oral squamous cell carcinoma. Indian Journal of Dental Research, 29(3), p.291.
29. Aigner T. Apoptosis, necrosis, or whatever: how to find out what really happens? J. Pathol. (2002) 198: 1-4.
30. Teja, K.V., Ramesh, S. and Priya, V., 2018. Regulation of matrix metalloproteinase-3 gene expression in inflammation: A molecular study. Journal of conservative dentistry: JCD, 21(6), p.592.
31. Pandiar, D., Ramani, P., Krishnan, R.P. and Monica, K., 2021. Multifaceted multinucleated giant cells in oral squamous cell carcinoma. Oral Oncology, 121, pp.105400-105400.
32. Yuwanati, M., Ramadoss, R., Kudo, Y., Ramani, P. and Senthil Murugan, M., 2022. Prevalence of oral submucous fibrosis among areca nut chewers: A systematic review and meta‐analysis. Oral Diseases.
33. Muthukrishnan, L., 2022. TGM3-A novel biomarker as a potential diagnostic target for head and neck squamous cell carcinoma (HNSCC). Oral oncology, 134, p.106118.
34. Jabin, Z., Nasim, I. and Agarwal, N., 2022. Comparative Evaluation of Salivary Fluoride Concentration after Topical Application of Silver Diamine Fluoride and Sodium Fluoride: A Randomized Controlled Trial. International Journal of Clinical Pediatric Dentistry, 15(3), p.371.