阿托伐醌通过ROS介导的自噬抑制结直肠癌细胞增殖的研究

Inhibitory effect of Atovaquone on the Proliferation of Colorectal Cancer Cells by ROS-mediated Autophagy

目的研究阿托伐醌抑制结直肠癌细胞增殖的作用机制。方法通过MTT法和细胞克隆实验检测阿托伐醌对结直肠癌细胞增殖的影响;免疫荧光、免疫印迹和流式细胞术检测阿托伐醌处理结直肠癌细胞后凋亡、自噬水平的变化以及胞内活性氧(ROS)的产生。结果阿托伐醌对结直肠癌细胞的增殖有浓度依赖性抑制作用,但凋亡水平无明显变化;阿托伐醌可促进结直肠癌细胞自噬,激活胞内ROS的产生;且抑制自噬可削弱阿托伐醌对结直肠癌增殖的抑制;同时,ROS清除剂NAC能抑制阿托伐醌诱导的结直肠癌细胞自噬。结论阿托伐醌通过促进结直肠癌细胞胞内ROS的产生诱导细胞自噬,进而抑制结直肠癌细胞增殖。

Objective To study the mechanism of atovaquone in inhibiting the proliferation of colorectal cancer cells. Methods The MTT method and cell cloning assay were used to detect the effect of atovaquone on the proliferation of colorectal cancer cells. The methods of Immunofluorescence, Western blotting and flow cytometry were used to detect the levels of apoptosis and autophagy and the production of intracellular reactive oxygen species (ROS) in the colorectal cancer cells treated with atovaquone. Results Atovaquone inhibited the proliferation of colorectal cancer cells in a concentration-dependent manner, but there was no significant changes in the apoptotic level. Atovaquone promoted the autophagy and activate the intracellular ROS production in colorectal cancer cells, and the inhibition of autophagy attenuated the inhibitory effect of atovaquone on the proliferation of colorectal cancer. Meanwhile, the ROS scavenger NAC inhibited the atovaquone- induced autophagy of colorectal cancer cells. Conclusion Atovaquone can induce autophagy by promoting the intracellular ROS production in colorectal cancer cells and thereby inhibit the proliferation of colorectal cancer cells.