miR-140-3p通过靶向MAPK1影响宫颈癌细胞增殖及顺铂敏感性的研究

Investigation of miR-140-3p targeting MAPK1 to influence cervical cancer cell proliferation and cisplatin sensitivity

目的研究miR-140-3p在宫颈癌细胞增殖和对顺铂敏感性中的作用及其机制。方法采用原位杂交技术检测宫颈癌及其癌旁组织中miR-140-3p的表达水平,并分析与临床病理参数的关系。通过CCK8实验、细胞克隆形成和细胞划痕实验,比较各组宫颈癌细胞增殖和迁移的差异。在不同浓度顺铂处理下,检测各组细胞的增殖、克隆形成及死亡率变化。利用miRNA数据库预测目标基因,并通过荧光素酶报告基因实验、qRT-PCR和Western blot检测目标基因mRNA和蛋白水平,以及凋亡相关蛋白表达,以验证miR-140-3p的潜在靶点。结果宫颈癌组织中miR-140-3p表达显著低于癌旁组织(P<0.05)。上调miR-140-3p可显著抑制宫颈癌细胞的增殖、克隆形成及迁移。CCK8、细胞克隆形成和划痕实验结果表明,不同浓度顺铂处理后,miR-140-3p过表达组MAPK1的表达下降,宫颈癌细胞增殖及克隆形成下降,而细胞凋亡增加(P<0.05)。结论miR-140-3p可能通过调控MAPK1基因抑制宫颈癌细胞增殖,并增强对顺铂的敏感性。

Objective To investigate the role and mechanism of miR-140-3p in the proliferation of cervical cancer cells and their sensitivity to cisplatin.Methods In situ hybridization was used to detect the expression levels of miR-140-3p in cervical cancer tissues and adjacent normal tissues,followed by an analysis of its correlation with clinicopathological parameters.CCK8 assay,cell colony formation,and scratch wound healing assays were performed to compare the differences in proliferation and migration among different groups of cervical cancer cells.Cells were treated with varying concentrations of cisplatin,and differences in proliferation,colony formation,and mortality rates were assessed.Target genes were predicted using miRNA databases,and luciferase reporter assays,qRT-PCR,and Western blotting were employed to detect the mRNA and protein levels of target genes as well as the expression of apoptosis-related proteins,thus validating the potential targets of miR-140-3p.Results miR-140-3p expression was significantly lower in cervical cancer tissues compared to adjacent normal tissues(P<0.05).Overexpression of miR-140-3p markedly inhibited the proliferation,colony formation,and migration of cervical cancer cells.CCK8,colony formation,and scratch wound healing assays indicated that with different concentrations of cisplatin,the miR-140-3p overexpression group showed inhibition effect of proliferation and clone formation of cervical cancer cells,and promotion effect of cell apoptosis by down regulation of the expression of MAPK1(P<0.05).Conclusion miR-140-3p may inhibit cervical cancer cell proliferation and enhance the sensitivity to cisplatin by regulating the MAPK1 gene expression.