香加皮杠柳苷通过调控circFMN2表达对宫颈癌HeLa细胞增殖及凋亡的影响

The effect of periplocin from cortex periplocae on the proliferation and apoptosis of cervical cancer HeLa cells by regulating the expression of circFMN2

目的 探讨香加皮杠柳苷(CPP)对宫颈癌HeLa细胞增殖、凋亡的影响及其对circFMN2的调控作用。方法 体外培养人宫颈癌细胞HeLa,加入不同剂量的CPP处理细胞,分别将si-NC、si-circFMN2转染至HeLa细胞,分别将pcDNA、pcDNA-circFMN2转染至HeLa细胞后加入CPP处理细胞;采用MTT法检测细胞增殖抑制率;采用流式细胞术检测细胞凋亡率;采用qRT-PCR法检测癌旁组织、宫颈癌组织及HeLa细胞中circFMN2的表达量;Western blot法检测Ki-67、Bcl-2、Bax蛋白表达量。结果 CPP可显著提高细胞增殖抑制率、凋亡率及Bax蛋白水平(P<0.05),显著降低Ki-67、Bcl-2蛋白水平(P<0.05),且呈剂量依赖性;与癌旁组织比较,宫颈癌组织中circFMN2的表达水平显著升高(P<0.05);CPP可显著降低circFMN2的表达水平(P<0.05),且呈剂量依赖性;转染si-circFMN2可显著提高细胞增殖抑制率、凋亡率及Bax蛋白水平(P<0.05),降低Ki-67、Bcl-2蛋白水平(P<0.05);转染pcDNA-circFMN2可明显逆转CPP对宫颈癌细胞的增殖和凋亡作用。结论 香加皮杠柳苷可通过抑制circFMN2的表达而抑制宫颈癌细胞增殖并促进细胞凋亡。

Objective To explore the effect of periplocin from cortex periplocae(CPP) on the proliferation and apoptosis of cervical cancer HeLa cells and its regulation of circFMN2.Methods Human cervical cancer cells HeLa were cultured in vitro,and different doses of CPP-treated cells were added.The si-NC and si-circFMN2 were transferred into HeLa cells respectively.After transfecting pcDNA and pcDNA-circFMN2 into HeLa cells,CPP treated cells were added.The MTT method was used to detect the inhibition rate of cell proliferation.Flow cytometry was used to detect the apoptosis rate.The qRT-PCR method was used to detect the expression of circFMN2 in adjacent tissues,cervical cancer tissues and HeLa cells.Western blot method was used to detect the expression of Ki-67,Bcl-2 and Bax protein.Results CPP could significantly increase cell proliferation inhibition rate,apoptosis rate and the protein level of Bax(P<0.05).And the protein levels of Ki-67and Bcl-2(P<0.05) in a dose-dependent manner were reduced.Compared with adjacent tissues,the expression level of circFMN2 in cervical cancer tissues significantly increased(P<0.05).CPP could significantly reduce the expression level of circFMN2(P<0.05) in a dose-dependent manner.Transfection of si-circFMN2 could significantly increase cell proliferation inhibition rate,apoptosis rate and the protein level of Bax(P<0.05).The protein levels of Ki-67 and Bcl-2(P<0.05) were reduced.Transfection of pcDNA-circFMN2 could obviously reverse the effect of CPP on the proliferation and apoptosis of cervical cancer cells.Conclusion Periplocin from cortex periplocae could inhibit the proliferation of cervical cancer cells and promote cell apoptosis by inhibiting the expression of circFMN2.