摘要
目的:探讨溶血磷脂酸受体4(LPAR4)在乳腺癌组织中的表达水平及与乳腺癌发生发展的关系,探究lncRNA Xist/miR-215-5p对LPAR4的调控作用。方法:通过Targetscan和Starbase预测LPAR4的上游通路lncRNA Xist/miR-215-5p。使用慢病毒转染LPAR4和miR-215-5p;使用质粒转染lncRNA Xist。在MDA-MB-231细胞系中,通过MTT和划痕实验检测细胞的增殖和转移能力。结果:LPAR4在乳腺癌细胞系中高表达,miR-215-5p和lncRNA Xist为低表达。Western blot显示LPAR4在乳腺癌细胞系中高表达。MTT和划痕实验提示上调LPAR4可以促进MDA-MB-231细胞的增殖和转移能力。进一步实验发现lncRNA Xist可以上调miR-215-5p,并且负性调控LPAR4;且lncRNA Xist可以抑制MDA-MB-231细胞的增殖能力。除此之外,在lncRNA Xist下调细胞中,上调miR-215-5p可以抑制lncRNA Xist下调介导的细胞增殖能力增强。结论:lncRNA Xist/miR-215-5p可能通过调控LPAR4的表达,为治疗乳腺癌提供可能的新靶点。LPAR4在乳腺癌细胞中高表达,可能成为乳腺癌潜在的肿瘤标志物。
Objective: To evaluate the expression of LPAR4 in breast cancer tissues and its association with breast cancer progression,and the regulating role of lncRNA Xist/miR-215-5p for LPAR4. Methods: The expression of LPAR4,lncRNA Xist and miR-215-5p in breast cancer cell line was tested by qRT-PCR. LPAR4 and miR-215-5p lentivirus and Xist plasmid were transfected. In MDA-MB-231 cell line,MTT and wound healing tests were used to investigate the proliferation and migration ability. Results: qRT-PCR indicated that LPAR4 expressed higher in breast cancer cell line,and miR-215-5p and Xist were expressed lower. MTT and wound healing test indicated that up regulation of LPAR4 could promote the proliferation and migration of MDA-MB-231 cell line. Furthermore,Xist could up regulate miR-215-5p and down regulate LPAR4,and Xist could inhibit the proliferation of MDA-MB-231. In addition,in Xist down regulated cell,up regulation of miR-215-5p could inhibit the Xist-associated enhanced cell proliferation. Conclusion: LPAR4 was associated with poorer prognosis in breast cancer and had the potential to be a novel biomarker. lncRNA Xist/miR-215-5p could provide novel therapeutic targets for breast cancer by targeting LPAR4.
引文
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