三黄煎剂降低ROS促进乳腺癌他莫昔芬耐药细胞凋亡的实验研究

San Huang decoction reduced ROS to promote MCF-7 tamoxifen-resistant cell apoptosis

目的探讨三黄煎剂对MCF-7他莫昔芬耐药(MCF-7/TAMR)细胞ROS水平和凋亡的影响。方法建立MCF-7/TAMR细胞系,采用CCK-8法检测他莫昔芬对乳腺癌细胞MCF-7、MCF-7/TAMR增殖的影响,并检测耐药指数;同时检测三黄煎剂对MCF-7/TAMR增殖的影响。免疫荧光法评价MCF-7、MCF-7/TAMR及各给药组的ROS的水平;流式细胞仪检测各实验组细胞凋亡情况;Western Blot法检测三黄煎剂对MCF-7/TAMR细胞凋亡相关蛋白表达对的影响。结果成功构建了MCF-7/TAMR细胞株,不仅在形态上发生了改变,而且对他莫昔芬的抵抗能力得到了增强,其RI指数为(3.61±0.35)(P<0.05),三黄煎剂对MCF-7/TAMR细胞株有明显的抑制作用,IC50值为(4.28±0.06)mg/m L,能够降低耐药细胞株的ROS水平,同时能够下调MCF-7/TAMR细胞的Bcl-2活性,上调Ba X/Bak,促使MCF-7/TAMR细胞产生凋亡。结论三黄煎剂通过降低乳腺癌MCF-7TAMR细胞内的ROS水平,抑制乳腺癌MCF-7TAMR细胞增殖并促进乳腺癌MCF-7TAMR细胞凋亡。

Objective To explore the effect of San Huang decoction on the ROS level and the apoptosis of MCF-7 tamoxifen resistance（ MCF-7 / TAMR）. Methods To establish MCF-7 / TAMR cell line,CCK 8 method testing tamoxifen for MCF-7,the influence of MCF-7 / TAMR proliferation,and detection of drug resistance index; While detecting San Huang decoction of MCF-7 / TAMR proliferation. Immunofluorescence method to evaluate MCF-7,MCF-7 /TAMR and each dose group of ROS levels; Flow cytometry instrument for detecting various experimental cell apoptosis; Western Blot method to detect San Huang decoction of MCF-7 / TAMR apoptosis related proteins expression of influence. Results The successful build the MCF-7 / TAMR cell lines,not only changed on the form,but also for tamoxifen resistance ability has been enhanced,the RI index was（ 3. 61 ± 0. 35）（ P〈0. 05）,San Huang decoction of MCF-7 / TAMR cell lines have obvious inhibitory effect,IC50 alue concentration of（ 4. 28 ± 0. 06） mg / m L,can reduce the ROS levels of drug resistance cell line,at the same time can cut MCF-7 the Bcl-2 / TAMR cell activity,increase Ba X / Bak,prompting MCF-7 / TAMR cells apoptosis. Conclusion San Huang decoction by reducing the ROS level in the MCF-7 tamr breast cancer cells,inhibit the proliferation and promote its apoptosis.