百里醌联合5-氟尿嘧啶对胃癌HGC-27细胞增殖和凋亡的影响

摘要

Objective To investigate the proliferation and apoptosis induced by thymoquinone(TQ) combine with 5 fluorouracil(5-Fu) on gastric cancer HGC-27 cells. Methods We conducted the experiment in central laboratory of Renmin Hospital of Wuhan University since September 2016. HGC-27 cells were treated with 50 μmol/L TQ,75 μg/ml 5 Fu or 50 μmol/L TQ + 75 μg/ml 5-Fu,and then the proliferation were detected by CCK-8. Apoptosis was detected by Hoechst stai-ning and Flow cytometry. The protein expressions of Bax and Bcl-2 were detected by Western blot. Results CCK-8 results showed that inhibited exposure to thymoquinone for 24 h clearly inhibited the viability of cells. The inhibition rate of the com-bined group was higher than 5-Fu group and TQ group(F=767.05,P<0.01). Hoechst result showed that the apoptosis rate in the combined group was significantly higher than the 5-Fu. Flow cytometry showed the apoptosis rate of the control group was (2.26 ± 0.17) % and the apoptosis rate of 50 μmol/L TQ,75 μg/ml 5-Fu and 50 μmol/L TQ + 75 ug/ml 5 Fu was (3.76 ± 0.44) %,(6.24 ± 0.19) % and(9.76 ± 0.25) % respectively (F=449.58,P<0.01). The protein expression of Bax was elevated (F= 58.803,P<0.01) and Bcl-2 was decreased (F=67.203, P<0.01) in combined group com-pared with single group. Conclusion Thymoquinone combined with 5 fluorouracil inhibits the proliferation and apoptosis in HGC-27 cell line,and thymoquinone can sensitize 5-Fu mediated growth inhibition.%目的 观察百里醌(TQ) 联合5-氟尿嘧啶(5-Fu) 诱导对胃癌HGC-27细胞发生增殖和凋亡的影响.方法 于2015年9月—2017年8月在武汉大学中心实验室进行实验,以TQ 50 μmol/L(TQ组)、5-Fu 75 μg/ml(5-Fu组)及 TQ 50 μmol/L + 5-Fu 75 μg/ml(联合组)刺激胃癌HGC-27细胞24 h后,CCK-8检测其对增殖的影响,Hoechst染色、流式细胞术检测细胞凋亡,蛋白质印迹法分析凋亡相关蛋白 Bax和Bcl-2的表达.结果 CCK-8结果显示,百里醌可抑制胃癌HGC-27细胞的增殖,联合组对细胞的抑制率高于5-Fu组及TQ组( F =767.05,P<0.01).Hoechst结果显示,联合组的凋亡率明显高于5-Fu组.流式细胞仪检测显示,对照组的凋亡率为(2.26 ± 0.17)%,TQ组、5-Fu组及联合组的凋亡率分别为(3.76 ± 0.44)%、(6.24 ± 0.19)%及(9.76 ± 0.25)%,联合组的凋亡率高于TQ组和5-Fu组( F =449.58,P<0.01).蛋白质印迹法结果显示,联合组的促凋亡蛋白Bax表达量均较(TQ组或5-Fu组)升高( F =58.803,P<0.01),且抑凋亡蛋白Bcl-2较TQ组或5-Fu组降低( F =67.203, P<0.01).结论 百里醌联合5-氟尿嘧啶可抑制胃癌HGC-27细胞增殖并促进其凋亡,百里醌可增加胃癌HGC-27细胞对5-氟尿嘧啶的敏感性.