circ_0023404通过靶向miR-153调控宫颈癌细胞增殖、侵袭迁移、凋亡及EMT

circ_0023404 regulates cervical cancer cell proliferation,invasion and migration,apoptosis and EMT by targeting miR-153

目的 探讨环状RNA 0023404(circ_0023404)是否靶向miR-153调控宫颈癌细胞增殖、侵袭迁移、凋亡及上皮间充质转化(EMT)。方法 实时荧光定量PCR(RT-qPCR)分析circ_0023404和miR-153在宫颈癌组织、癌旁组织中表达水平。CCK-8法、流式细胞术、划痕愈合、Transwell实验分析circ_0023404和miR-153对宫颈癌SW756细胞增殖、凋亡、迁移和侵袭的影响。双萤光素酶报告基因实验、RT-qPCR用于确定circ_0023404和miR-153相互作用。蛋白质印迹法(Western blot)分析Ki-67、基质金属蛋白酶2(MMP-2)、MMP-9、上皮细胞钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)、B细胞淋巴瘤/白血病-2(Bcl-2)以及Bcl相关x蛋白(Bax)表达。结果 宫颈癌组织中circ_0023404表达高于癌旁组织,miR-153表达低于癌旁组织(P<0.05)。敲低circ_0023404表达或上调miR-153表达可显著降低SW756细胞活力、迁移和侵袭能力,增加细胞凋亡率,上调Bax和E-cadherin蛋白,并下调Bcl-2、MMP-2、MMP-9以及Vimentin蛋白表达(P<0.05)。circ_0023404与miR-153直接结合并负调控miR-153表达。下调miR-153表达可显著减弱敲低circ_0023404对SW756细胞增殖、凋亡、迁移、侵袭以及相关蛋白表达的影响(P<0.05)。结论 敲低circ_0023404可能通过上调miR-153表达抑制宫颈癌细胞增殖、EMT、迁移和侵袭,并诱导细胞凋亡。

Objective To investigate whether circular RNA 0023404(circ_0023404)targets miR-153 to regulate cervical cancer cell proliferation,invasion and migration,apoptosis and epithelial-mesenchymal transition(EMT).Methods Real-time fluorescence quantitative PCR(RT-qPCR)was used to analyze the expression levels of circ_0023404 and miR-153 in cervical cancer tissues and adjacent tissues.CCK-8,flow cytometry,scratch healing and Transwell assay were used to analyze the effects of circ_0023404 and miR-153 expression on the proliferation,apoptosis,migration and invasion of cervical cancer SW756 cells.The dual luciferase reporter gene experiment and RT-qPCR were performed to determine the interaction between circ_0023404 and miR-153.The expression of Ki-67,matrix metalloproteinase 2(MMP-2),MMP-9,epithelial cell cadherin(E-cadherin),Vimentin(Vimentin),B-cell lymphoma/leukemia-2(Bcl-2)and Bcl-related protein x(Bax)expression was analysed by Western blot.Results The expression of circ_0023404 in cervical cancer tissue was higher than that in adjacent tissues,whereas the expression of miR-153 was lower than that in adjacent tissues(P<0.05).Down-regulating circ_0023404 expression or up-regulating miR-153 expression could significantly reduce cell viability,migration and invasion ability,increase cell apoptosis rate,up-regulate the expression of Bax and E-cadherin proteins,and down-regulate the expression of Bcl-2,MMP-2,MMP-9 and Vimentin proteins(P<0.05).circ_0023404 directly binds to miR-153 and negatively regulates the expression of miR-153.Down-regulation of miR-153 expression could significantly reverse the effects of circ_0023404 inhibition on SW756 cell proliferation,apoptosis,migration,invasion and related protein expression(P<0.05).Conclusion Knockdown of circ_0023404 may inhibit the proliferation,EMT,migration and invasion of cervical cancer cells and induce cell apoptosis through up-regulation ofmiR-153 expression.