二甲双胍依赖ARID1A促进子宫内膜腺癌细胞发生有丝分裂灾难

Metformin relies on ARID1A to promote mitotic catastrophe in endometrial adenocarcinoma cells

目的建立ARID1A稳定敲除的人子宫内膜腺癌KLE细胞株,初步探讨二甲双胍抑制子宫内膜腺癌细胞增殖的作用机制。方法利用CRISPR/Cas9技术构建ARID1A稳定敲除的人子宫内膜腺癌KLE细胞株;给予二甲双胍刺激后,Western blot检测KLE细胞ARID1A敲除前后Cyclin B1、p-H2AX以及Cleaved-caspase3蛋白表达水平;免疫荧光实验检测二甲双胍刺激下,KLE细胞ARID1A敲除前后多核细胞所占百分比;CCK8实验检测二甲双胍刺激下,KLE细胞ARID1A敲除前后的增殖水平。结果经筛选,成功构建ARID1A完全敲除的KLE细胞株;ARID1A敲除后,子宫内膜腺癌细胞对于二甲双胍造成的有丝分裂灾难、增殖抑制以及凋亡不敏感。结论二甲双胍依赖ARID1A导致子宫内膜腺癌细胞发生有丝分裂灾难。

Objective To establish KLE cell line of human endometrial adenocarcinomawith stable knockout of ARID1A and investigate the mechanism of Metformin inhibiting the proliferation of endometrial adenocarcinoma cells. Methods ARID1A stable knockout human endometrial adenocarcinoma KLE cell line was constructed using CRISPR/Cas9 technology. After stimulation with Metformin, the expressions of CyclinB1, p-H_2AX and Cleaved caspase3 were detected by Western blot. The percentage of Metformin-stimulated multinucleated cells before and after ARID1A knockout was detected by immunofluorescence. The proliferation level of metformin stimulated KLE cells before and after ARID1A knockout was detected by CCK8 assay. Results After screening, KLE cell lines with ARID1A completely knocked out were successfully constructed. After ARID1A knockout, endometrial adenocarcinoma cells were insensitive to mitosis disaster, proliferation inhibition, and apoptosis caused by Metformin. Conclusion Metformin caused mitotic catastrophe is ARID1A dependent in endometrial adenocarcinoma cells.