Effects of berberine on proliferation, cell cycle distribution and apoptosis of human breast cancer T47D and MCF7 cell lines

Document Type: Original Article

Authors

1 Molecular Research Lab, Department of Pharmacology and Toxicology, Tehran University of Medical Sciences, Tehran, Iran

2 Department of Molecular Medicine, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran.

3 Department of Pharmacology and Toxicology, Faculty of Pharmacy, Tehran University of Medical Sciences, Tehran, Iran

Abstract

Objective(s):Berberine, a naturally occurring isoquinoline alkaloid, has shown antitumor properties in some in vitro systems. But the effect of berberine on breast cancer has not yet been completely studied. In this study, we evaluated anticancer properties of berberine in comparison to doxorubicin.
Materials and Methods: The antiproliferative effects of berberine and doxorubicin alone and in combination were evaluated in T47D and MCF7 cell lines using MTT cytotoxicity assay. In addition, flow cytometry analysis was performed to evaluate the cell cycle alteration and apoptosis induction in these cell lines following exposure to berberine and doxorubicin alone and in combination.
Results: The IC50 of berberine was determined to be 25 µM after 48 hr of treatment in both cell lines but for doxorubicin it was 250 nM and 500 nM in T47D and MCF-7 cell lines, respectively. Co-treatment with berberine and doxorubicin increased cytotoxicity in T47D cells more significantly than in MCF-7 cells. Flow cytometry results demonstrated that berberine alone or in combination with doxorubicin induced G2/M arrest in the T47D cells, but G0/G1 arrest in the MCF-7 cells. Doxorubicin alone induced G2/M arrest in both cell lines. Furthermore, berberine and doxorubicin alone or in combination significantly induced apoptosis in both cell lines.
Conclusion: Berberine alone and in combination with doxorubicin inhibited cell proliferation, induced apoptosis and altered cell cycle distribution of breast cancer cells. Therefore, berberine showed to be a good candidate for further studies as a new anticancer drug in the treatment of human breast cancer.

Keywords


1. Razmkhah M, Ghaderi A. HLA Class I Allele Frequencies in Southern Iranian Women With Breast Cancer. Iran J Basic Med Sci 2013; 16:140-143.

2. Cao H, Yang Zh, Jiang G. Expression and clinical significance of activating transcription factor 3 in human breast cancer. Iran J Basic Med Sci 2013; 16:1151-1154.

3. Karimian Fathi N, Shekari Khaniani M, Montazeri V, Mansoori Derakhshan S. Minor role of BRCA2 mutation (Exon2 and Exon11) in patients with early-onset breast cancer amongst Iranian Azeri-Turkish women. Iran J Basic Med Sci 2014; 17:108-111.

4. Tavakoli-Yaraki M, Karami-Tehrani F. Apoptosis Induced by 13-S hydroxyoctadecadienoic acid in the Breast Cancer Cell Lines, MCF-7 and MDA-MB-231. Iran J Basic Med Sci 2013; 16: 661-669.

5. Lynch MD, Cariati M, Purushotham AD. Breast cancer, stem cells and prospects for therapy. Breast Cancer Res 2006; 8:211.

6. Zhong L, Shen H, Huang C, Jing H, Cao D. AKR1B10 induces cell resistance to daunorubicin and idarubicin by reducing C13 ketonic group. Toxicol Appl Pharmacol 2011; 255:40-47.

7. Moretti E, Oakman C, Di Leo A. Predicting anthra-cycline benefit: have we made any progress? Curr Opin Oncol 2009; 21:507-515.

8. Hussein MA. Preclinical rationale, mechanisms of action, and clinical activity of anthracyclines in myeloma. Clin Lymphoma Myeloma 2007; 4: S145-149.

9. Brechbuhl HM, Kachadourian R, Min E, Chan D, Day BJ. Chrysin enhances Doxorubicin-induced cytotoxicity in human lung epithelial cancer cell lines: the role of glutathione. Toxicol Appl Pharmacol 2012; 258:1-9.

10. Hembruff SL, Laberge ML, Villeneuve DJ, Guo B, Veitch Z, Cecchetto M, et al. Role of drug transporters and drug accumulation in the temporal acquisition of drug resistance. BMC Cancer 2008; 8:318.

11. Zhong L, Shen H, Huang C, Jing H, Cao D. AKR1B10 induces cell resistance to daunorubicin and idarubicin by reducing C13 ketonic group. Toxicol Appl Pharmacol 2011; 255:40-47.

12. Meeran SM, Katiyar S, Katiyar SK. Berberine-induced apoptosis in human prostate cancer cells is initiated by reactive oxygen species generation. Toxicol Appl Pharmacol 2008; 229:33-43.

13. Mantena SK, Sharma SD, Katiyar SK. Berberine, a natural product, induces G1-phase cell cycle arrest and caspase-3-dependent apoptosis in human prostate carcinoma cells. Mol Cancer Ther 2006; 5:296-308.

14. Peng PL, Hsieh YS, Wang CJ, Hsu JL, Chou FP. Inhibitory effect of berberine on the invasion of human lung cancer cells via decreased productions of urokinase-plasminogen activator and matrix metalloproteinase-2. Toxicol Appl Pharmacol 2006; 214:8-15.

15. Kim S, Han J, Lee SK, Choi MY, Kim J, Lee J, et al. Berberine suppresses the TPA-induced MMP-1 and MMP-9 expressions through the inhibition of PKC-alpha in breast cancer cells. J Surg Res 2012;              176:e21-9.

16. Kim JB, Ko E, Han W, Shin I, Park SY, Noh DY. Berberine diminishes the side population and ABCG2 transporter expression in MCF-7 breast cancer cells. Planta Med 2008; 74:1693-1700.

17. Patil JB, Kim J, Jayaprakasha GK. Berberine induces apoptosis in breast cancer cells (MCF-7) through mitochondrial-dependent pathway. Eur J Pharmacol 2010; 645:70-78.

18. Rezaei PF, Fouladdel S, Hassani S, Yousefbeyk F, Ghaffari SM, Amin G, et al. Induction of apoptosis and cell cycle arrest by pericarp polyphenol-rich extract  of Baneh in human colon carcinoma HT29 cells. Food Chem Toxicol 2012; 50:1054-1059.

19. Shahveisi K, Mousavi SH, Hosseini M, Khajavi Rad A, Jalali SA, Rajaei Z, et al. The role of local renin-angiotensin system on high glucose-induced cell toxicity, apoptosis and reactive oxygen species production in PC12 cells. Iran J Basic Med Sci 2014; 17:613-621.

20. Simstein R, Burow M, Parker A, Weldon C, Beckman B. Apoptosis, chemoresistance, and breast cancer: insights from the MCF-7 cell model system. Exp Biol Med (Maywood) 2003; 228: 995-1003.

21. Lonning PE, Knappskog S, Staalesen V, Chrisanthar R, Lillehaug JR. Breast cancer prognostication and prediction in the postgenomic era. Ann Oncol 2007; 18:1293-1306.

22. Bouchalova K, Cizkova M, Cwiertka K, Trojanec R, Hajduch M. Triple negative breast cancer--current status and prospective targeted treatment based on HER1 (EGFR), TOP2A and C-MYC gene assessment. Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub 2009; 153: 13-17.

23. Kuo CL, Chi CW, Liu TY. The anti-inflammatory potential of berberine in vitro and in vivo. Cancer Lett 2004; 203:127-137.

24. Kim S, Choi JH, Kim JB, Nam SJ, Yang JH, Kim JH, et al. Berberine suppresses TNF-alpha-induced MMP-9 and cell invasion through inhibition of AP-1 activity in MDA-MB-231 human breast cancer. Cells Molecules 2008; 13:2975-2985.

25. Ramasamy S, Abdul Wahab N, Zainal Abidin N, Manickam S. Effect of extracts from Phyllanthus watsonii Airy Shaw on cell apoptosis in cultured human breast cancer MCF-7 cells. Exp Toxicol Pathol 2012; 65:341-349.

26. Abd El-Wahab AE, Ghareeb DA, Sarhan EE, Abu-Serie MM, El Demellawy MA. In vitro biological assessment of berberis vulgaris and its active constituent, berberine: antioxidants, anti-acetylcholinesterase, anti-diabetic and anticancer  effects. BMC Complement Altern Med 2013; 13:218.

27. Kuo HP, Chuang TC, Tsai SC, Tseng HH, Hsu SC, Chen YC, et al. Berberine, an isoquinoline alkaloid, inhibits the metastatic potential of breast cancer cells via Akt pathway modulation. J Agric Food Chem 2012; 60:9649-9658.

28. Hill GM, Moriarity DM, Setzer WN. Attenuation of cytotoxic natural product DNA intercalating agents by caffeine. Sci Pharm 2011; 79:729-747.

29. Pinto-Garcia L, Efferth T, Torres A, Hoheisel JD, Youns M. Berberine inhibits  cell growth and mediates caspase-independent cell death in human pancreatic cancer cells. Planta Med 2010; 76:1155-1161.

30. Tsang CM, Cheung YC, Lui VW, Yip YL, Zhang G, Lin VW, et al. Berberine suppresses tumorigenicity and growth of nasopharyngeal carcinoma cells by inhibiting STAT3 activation induced by tumor associated fibroblasts. BMC Cancer 2013; 13:619. 

31. Pazhang Y, Ahmadian S, Javadifar N, Shafiezadeh M. COX-2 and survivin reduction may play a role in berberine-induced apoptosis in human ductal breast epithelial tumor cell line. Tumour Biol 2012; 33:207-214.

32. Kim JB, Lee KM, Ko E, Han W, Lee JE, Shin I, et al. Berberine  inhibits growth of the breast cancer cell lines MCF-7 and MDA-MB-231. Planta Med 2008; 74:39-42.

33. Xu LN, Lu BN, Hu MM, Xu YW, Han X, Qi Y, et al. Mechanisms involved in the cytotoxic effects of berberine on human colon cancer HCT-8 cells. Biocell  2012; 36:113-120.

34. Chidambara Murthy KN, Jayaprakasha GK, Patil BS. The natural alkaloid berberine targets multiple pathways to induce cell death in cultured human colon  cancer cells. Eur J Pharmacol 2012; 688:14-21.

35. Wu K, Yang Q, Mu Y, Zhou L, Liu Y, Zhou Q, et al. Berberine inhibits the proliferation of colon cancer cells by inactivating Wnt/β-catenin signaling. Int  J Oncol 2012; 41:292-298.

36. Lin JP, Yang JS, Lee JH, Hsieh WT, Chung JG. Berberine induces cell cycle arrest and apoptosis in human gastric carcinoma SNU-5 cell line. World J Gastroenterol 2006; 12:21-28.

37. Wang Y, Liu Q, Liu Z, Li B, Sun Z, Zhou H, et al. Berberine, a genotoxic alkaloid, induces ATM-Chk1 mediated G2 arrest in prostate cancer cells. Mutat Res 2012; 734:20-29.

38. Choi MS, Oh JH, Kim SM, Jung HY, Yoo HS, Lee YM,  et al. Berberine inhibits p53-dependent cell growth through induction of apoptosis of prostate cancer cells. Int J Oncol 2009; 34:1221-1230.

39. Park KS, Kim JB, Lee SJ, Bae J. Berberine-induced growth inhibition of epithelial ovarian carcinoma cell lines. J Obstet Gynaecol Res 2012; 38:535-540.

40. Park KS, Kim JB, Bae J, Park SY, Jee HG, Lee KE, et al. Berberine inhibited  the growth of thyroid cancer cell lines 8505C and TPC1. Yonsei Med J 2012; 53:346-351.

41. Yan K, Zhang C, Feng J, Hou L, Yan L, Zhou Z, et al. Induction of G1 cell cycle arrest and apoptosis by berberine in bladder cancer cells. Eur J Pharmacol 2011; 661:1-7.

42. Hou Q, Tang X, Liu H, Tang J, Yang Y, Jing X, et al. Berberine induces cell death in human hepatoma cells in vitro by downregulating CD147. Cancer Sci 2011; 102:1287-1292.

43. Wang XN, Han X, Xu LN, Yin LH, Xu YW, Qi Y, et al. Enhancement of apoptosis of human hepatocellular carcinoma SMMC-7721 cells through synergy of berberine and evodiamine. Phytomedicine 2008; 15:1062-1068.

44. Tsang CM, Lau EP, Di K, Cheung PY, Hau PM, Ching YP, et al. Berberine inhibits Rho GTPases and cell migration at low doses but induces G2 arrest and apoptosis at high doses in human cancer cells. Int J Mol Med 2009; 24:131-138.

45. Lin CC, Yang JS, Chen JT, Fan S, Yu FS, Yang JL, et al. Berberine induces apoptosis in human HSC-3 oral cancer cells via simultaneous activation of the death receptor-mediated and mitochondrial pathway. Anticancer Res 2007; 27:3371-3378.

46. Mantena SK, Sharma SD, Katiyar SK. Berberine inhibits growth, induces G1 arrest and apoptosis in human epidermoid carcinoma A431 cells by regulating Cdki-Cdk-cyclin cascade, disruption of mitochondrial membrane potential and cleavage of caspase 3 and PARP. Carcinogenesis 2006; 27:2018-2027.

47. Kuo CL, Chou CC, Yung BY. Berberine complexes with DNA in the berberine-induced apoptosis in human leukemic HL-60 cells. Cancer Lett 1995; 93:193-200.

48. Letasiová S, Jantová S, Miko M, Ovádeková R, Horváthová M. Effect of berberine on proliferation, biosynthesis of macromolecules, cell cycle and induction of intercalation with DNA, dsDNA damage and apoptosis in Ehrlich ascites carcinoma cells. J Pharm Pharmacol 2006; 58:263-270.

49. Kim JB, Yu JH, Ko E, Lee KW, Song AK, Park SY, et al. The alkaloid Berberine inhibits the growth of Anoikis-resistant MCF-7 and MDA-MB-231  breast cancer cell lines by inducing cell cycle arrest. Phytomedicine 2010; 17:436-440.

50. Mittal A, Tabasum S, Singh RP. Berberine in combination with Doxorubicin suppresses growth of murine melanoma B16F10 cells in culture and xenograft. Phytomedicine 2014; 21:340-347.

51. Yip NK, Ho WS. Berberine induces apoptosis via the mitochondrial pathway in liver cancer cells. Oncol Rep 2013; 30:1107-1112.  

52. He W, Wang B, Zhuang Y, Shao D, Sun K, Chen J. Berberine inhibits growth and induces G1 arrest and apoptosis in human cholangiocarcinoma QBC939 cells. J Pharmacol Sci 2012; 119:341-348.

53. Choi MS, Yuk DY, Oh JH, Jung HY, Han SB, Moon DC, et al. Berberine inhibits  human neuroblastoma cell growth through induction of p53-dependent apoptosis. Anticancer Res 2008; 28:3777-3784.

54. Eom KS, Hong JM, Youn MJ, So HS, Park R, Kim JM,              et al. Berberine induces G1  arrest and apoptosis in human glioblastoma T98G cells through mitochondrial/caspases

 

pathway. Biol Pharm Bull 2008; 31:558-562.

55. Serafim TL, Oliveira PJ, Sardao VA, Perkins E, Parke D, Holy J. Different concentrations of berberine result in distinct cellular localization patterns and cell cycle effects in a melanoma cell line. Cancer Chemother Pharmacol 2008; 6:1007-10018.

56. Lin HL, Liu TY, Wu CW, Chi CW. Berberine modulates expression of mdr1 gene product and the responses of digestive track cancer cells to Paclitaxel. Br J Cancer 1999; 81:416-422.

57. Hickman JA. Apoptosis induced by anticancer drugs. Cancer Metastasis Rev 1992; 11:121-139.

58. Crompton M. Bax, Bid and the permeabilization of the mitochondrial outer membrane in apoptosis. Curr Opin Cell Biol 2000; 12:414-419.