【摘要】 目的:研究环状RNA缺失-小同源异形-2样蛋白(CircASH2L)调节miR-128-3p/MAP激酶相互作用丝氨酸/苏氨酸激酶2(MKNK2)轴对卵巢癌细胞紫杉醇耐药性的影响。方法:实时荧光定量PCR检测人卵巢癌细胞株SKOV3和人卵巢癌紫杉醇耐药细胞株SKOV3-TR30中miR-128-3p与CircASH2L、MKNK2表达。体外培养SKOV3-TR30细胞，随机分为对照组、紫杉醇组、紫杉醇+阴性对照组、紫杉醇+CircASH2L敲低组、紫杉醇+CircASH2L敲低+miR-128-3p inhibitor组，分组处理后，检测各组细胞增殖率、凋亡率，miR-128-3p及CircASH2L、MKNK2、多重耐药1(MDR1)、多药耐药相关蛋白5(MRP5)mRNA表达，MKNK2及P-gp、MRP5蛋白表达。双荧光素酶报告基因实验检测SKOV3-TR30中CircASH2L对miR-128-3p的靶向调节及miR-128-3p对MKNK2的靶向调节。结果:与SKOV3细胞相比，SKOV3-TR30细胞中CircASH2L与MKNK2 mRNA表达升高，miR-128-3p表达降低，差异均有统计学意义(P<0.05)。与对照组、紫杉醇组分别相比，紫杉醇+CircASH2L敲低组的细胞增殖率降低，CircASH2L及MKNK2、MDR1、MRP5 mRNA表达降低，MKNK2及P-gp、MRP5蛋白表达降低，细胞凋亡率、miR-128-3p表达均升高，差异均有统计学意义(P<0.05);紫杉醇组、紫杉醇+阴性对照组细胞各指标无明显变化(P>0.05)。下调miR-128-3p可逆转敲低CircASH2L对紫杉醇处理下SKOV3-TR30细胞各指标变化。CircASH2L可靶向下调SKOV3-TR30中miR-128-3p且miR-128-3p可靶向下调MKNK2。结论:敲低CircASH2L可通过促进miR-128-3p表达而下调MKNK2表达，进而抑制卵巢癌细胞的紫杉醇耐药性。更多还原

【Abstract】 Objective:To study the impact of circular RNA absent-small-homeotic-2-like protein(CircASH2L) on paclitaxel resistance in ovarian cancer cells by regulating miR-128-3p/MAP kinase interaction serine/threonine kinase 2(MKNK2) axis.Methods:Real-time quantitative PCR was used to detect the expression levels of miR-128-3p, CircASH2L and MKNK2 in human ovarian cancer cell line SKOV3 and human ovarian cancer paclitaxel-resistant cell line SKOV3-TR30.SKOV3-TR30 cells were cultured in vitro and randomly grouped into control group, paclitaxel group, paclitaxel + negative control group, paclitaxel + CircASH2L knockdown group, and paclitaxel + CircASH2L knockdown + miR-128-3p inhibitor group.After grouping, to detect cell proliferation rate, the apoptosis rate, the levels of miR-128-3p, CircASH2L,MKNK2 and multidrug resistance 1(MDR1),multidrug resistance-related protein 5(MRP5) mRNA expression, the protein expression levels of MKNK2,P-gp and MRP5 of cells in each group.The targeted regulation on miR-128-3p by CircASH2L and the targeted regulation on MKNK2 by miR-128-3p in SKOV3-TR30 were detected by dual-luciferase reporter gene assay.Results:Compared with SKOV3 cells, the expression levels of CircASH2L and MKNK2 mRNA in SKOV3-TR30 cells were increased(P<0.05),the expression of miR-128-3p was decreased(P<0.05).Compared with the control group and the paclitaxel group, the cell proliferation rate, CircASH2L and MKNK2,MDR1,MRP5 mRNA expressions, MKNK2,P-gp, and MRP5 protein expressions in the paclitaxel + CircASH2L knockdown group were decreased(P<0.05),the apoptosis rate and the expression of miR-128-3p were increased(P<0.05).There was no significant change in each index of cells in the paclitaxel group and the paclitaxel + negative control group(P>0.05).Down-regulation of miR-128-3p can reverse the changes of various indicators of SKOV3-TR30 cells treated with paclitaxel by knocking down CircASH2L.CircASH2L can target down miR-128-3p in SKOV3-TR30 and miR-128-3p can target down MKNK2.Conclusion:Knockdown of CircASH2L can down-regulate MKNK2 by promoting the expression of miR-128-3p, thereby inhibiting the paclitaxel resistance of ovarian cancer cells.更多还原