咪达唑仑通过下调circ_0000285表达影响宫颈癌CaSki细胞增殖和凋亡的实验研究

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摘要 目的：探究咪达唑仑对宫颈癌CaSki细胞增殖和凋亡的影响及可能机制。方法：用不同剂量（0、15、30、60 μg/mL）咪达唑仑干预CaSki细胞,CCK-8法、流式细胞术分别测定细胞增殖活性和凋亡,蛋白质印迹法测定Ki-67、pro-caspase3、cleaved-caspase3蛋白表达,RT-qPCR法测定circ_0000285表达。通过转染circ_0000285小干扰RNA或过表达载体构建干扰circ_0000285表达或过表达circ_0000285的CaSki细胞,再用0或60 μg/mL咪达唑仑干预,上述相同方法测定细胞增殖活性、凋亡及相关蛋白（Ki-67、pro-caspase3、cleaved-caspase3）表达。结果：CaSki细胞经咪达唑仑干预后,增殖活性、Ki-67、pro-caspase3蛋白表达量降低,细胞凋亡率、cleaved-caspase3蛋白表达量升高,且细胞中circ_0000285表达量降低。干扰circ_0000285表达后,CaSki细胞增殖活性、Ki-67、pro-caspase3蛋白表达量降低,细胞凋亡率、cleaved-caspase3蛋白表达量升高。过表达circ_0000285逆转咪达唑仑对CaSki细胞增殖、凋亡的影响。结论：咪达唑仑可能通过下调circ_0000285抑制宫颈癌CaSki细胞增殖,并促进细胞凋亡。

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	张江川
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	张春梅

关键词 ：宫颈癌, 咪达唑仑, circ_0000285, 细胞增殖, 凋亡

Abstract：Objective To explore the effect and possible mechanism of midazolam on the proliferation and apoptosis of cervical cancer CaSki cells. Methods CaSki cells were intervened with different doses（0,15,30,60 μg/mL）of midazolam,and the cell proliferation activity and apoptosis were measured by CCK-8 method and flow cytometry,respectively. The protein expressions of pro-caspase3 and cleaved-caspase3 were determined by western blotting,and the expression of circ_0000285 was determined by RT-qPCR. After CaSki cells that interfered with circ_0000285 expression or overexpressed circ_0000285 were constructed by transfecting circ_0000285 small interfering RNA or overexpression vector,they were intervened with 0 or 60 μg/mL midazolam,and then the same method sas above were used to measure cell proliferation activity,apoptosis and the expression of related proteins（Ki-67,pro-caspase3,cleaved-caspase3）. Results After CaSki cells were intervened by midazolam,the proliferation activity and the protein expressions of Ki-67 and pro-caspase3 were decreased,but the apoptosis rate and the protein expressions of cleaved-caspase3 were increased,and the expression of circ_0000285 in the cells was decreased. After interfering the expression of circ_0000285,the proliferation activity and the protein expressions of Ki-67 and pro-caspase3 in CaSki cells were decreased,but the apoptosis rate and the protein expressions of cleaved-caspase3 were increased. Overexpression of circ_0000285 reversed the effects of midazolam on the proliferation and apoptosis of CaSki cells. Conclusion Midazolam may inhibit the proliferation of cervical cancer CaSki cells and promote cell apoptosis by down-regulating circ_0000285.

Key words： cervical cancer midazolam circ_0000285 cell proliferation apoptosis

收稿日期: 2022-09-07

中图分类号:	R737.33
	R363

通讯作者: 张春梅,E-mail：452335757@qq.com

引用本文:

张江川, 吴丹, 蒋伟, 张春梅. 咪达唑仑通过下调circ_0000285表达影响宫颈癌CaSki细胞增殖和凋亡的实验研究[J]. 湖南师范大学学报(医学版), 2023, 20(5): 38-43. ZHANG Jiangchuan, WU Dan, JIANG Wei, ZHANG Chunmei. Experimental study on the effect of midazolam on the proliferation and apoptosis of cervical cancer CaSki cells by down-regulating the expression of circ_0000285. HuNan ShiFan DaXue XueBao(YiXueBan), 2023, 20(5): 38-43.

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http://yxb.hunnu.edu.cn/CN/ 或 http://yxb.hunnu.edu.cn/CN/Y2023/V20/I5/38

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