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| Study on the effect of Xiaotan Sanjie Recipe on apoptosis of colon cancer stem-like cells and its related mechanism |
| ZHOU Yuqi1,2 ZHANG Yingcheng1 WEI Pinkang1▲ YE Min1▲ |
1.Department of Traditional Chinese Medicine, Changzheng Hospital, the Second Military Medical University, Shanghai 200003, China;
2.Department of Hematology and Oncology, the 101st Hospital of People's Liberation Army, Jiangsu Province, Wuxi 214044, China |
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Abstract Objective To investigate effect of Xiaotan Sanjie Recipe on apoptosis of colon cancer stem-like cells (CCSCs) and its related mechanism. Methods The CCSCs were induced from colon cancer cell line HCTl16 in serum-free medium. The expressions of CD44 were detected to identify CCSCs. The lyophilized powder of Xiaotan Sanjie Recipe Extract was dissolved in cell culture fluid, so as to make preserving fluid of Xiaotan Sanjie Recipe. CCSCs were treated with different concentrations of Xiaotan Sanjie Recipe (0.125, 0.25, 0.5, 1.0 mg/mL) for 72 h. The effects of different concentrations of Xiaotan Sanjie Recipe for CCSCs were detected by Annexin-V/PI staining combined with flow cytometry. The expression changes of Bax, Bcl-2 protein after drug intervention were detected by Western blot. Results HCT116 cells formed cancer stem cell spheres in serum-free medium. The proportion of CD44 cells in cell spheres was (49.69±1.89)%. Xiaotan Sanjie Recipe could induce the cell apoptosis, the apoptosis rate was statistically significant different compared with the control group (0 mg/mL) (P ＜ 0.05), the apoptotic effects showed concentration-dependent. After application of Xiaotan Sanjie Recipe, the expression of pro-apoptotic protein Bax of Bcl-2 family was increased, meanwhile the expression of anti-apoptotic protein Bcl-2 was decreased. The effect was more pronounced in 0.5 mg/mL and 1.0 mg/mL concentration groups. Conclusion CCSCs are successfully induced from HCT116 colon cancer cell line. Xiaotan Sanjie Recipe can induce CCSCs in a dose-dependent manner, the mechanism may be related to the increased expression of pro-apoptotic protein Bax and decreased expression of anti-apoptotic protein Bcl-2.
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