miR-129通过靶向调控HMGA2抑制卵巢癌细胞侵袭迁移的作用机制

Mechanism of miR-129 inhibiting invasion and migration of ovarian cancer cells by targeting HMGA2

目的:研究miR-129在高级别浆液性卵巢癌(high-grade serous ovarian cancer,HGSOC)组织及细胞系中的表达,并探讨其对卵巢癌细胞侵袭迁移的影响及作用机制。方法:实时荧光定量PCR(real-time quantitative PCR,qRT-PCR)检测miR-129,HMGA2在70例HGSOC组织和30例正常输卵管组织中的表达,以及miR-129在人卵巢癌细胞系和人正常卵巢上皮细胞系中的表达;Pearson相关分析评估HGSOC组织中miR-129与HMGA2表达的相关性;将miR-129 mimic和对照转染卵巢癌细胞CAOV后,用Transwell小室法检测细胞侵袭情况,细胞划痕试验检测细胞迁移情况、HMGA2 mRNA和蛋白表达变化;采用生物信息软件及荧光素酶报告基因试验。分析miR-129对HMGA2基因的靶向性作用;转染HMGA2过表达质粒,观察HMGA2对miR-129 mimic CAOV细胞侵袭和迁移的影响。结果:miR-129在HGSOC组织和细胞系中的表达均显著低于正常输卵管组织和正常卵巢上皮细胞(均P<0.05);HMGA2在HGSOC组织中的表达显著高于正常输卵管组织(均P<0.05),与miR-129的表达呈负相关(r=.0.712,P<0.05);与对照组相比,miR-129 mimic组CAOV细胞侵袭迁移能力下降,HMGA2 mRNA和蛋白表达减少(均P<0.05);生物信息软件预测和荧光素酶报告基因实验显示HMGA2为miR-129的靶基因;与miR-129+对照组相比,miR-129 mimic+HMGA2组细胞侵袭迁移能力明显升高(均P<0.05)。结论:miR-129在HGSOC组织中低表达,可能通过靶向下调HMGA2抑制卵巢癌细胞的侵袭迁移。

Objective: To investigate the expression of miR-129 in high-grade serous ovarian cancer(HGSOC) tissues and cell lines, and to explore its effect on the invasion and migration of ovarian cancer cells and its mechanism. Methods: Real-time quantitative PCR(qRT-PCR) was used to detect the expression of miR-129 and HMGA2 in 70 HGSOC tissues and 30 normal fallopian tubes, and the expression of miR-129 in human ovarian cancer cell lines and human normal ovarian epithelial cells. Pearson correlation analysis evaluated the correlation between miR-129 and HMGA2 expression in HGSOC tissues. After transfection of ovarian cancer cells CAOV with a miR-129 mimic and control, cell invasion was detected by Transwell microscopy, cell migration was detected by cell scratch test, and HMGA2 mRNA and protein expression changes were detected. Bioinformatics software and luciferase reporter gene assay were used to analyze the targeting effect of miR-129 on HMGA2 gene. HMGA2 overexpression plasmid was transfected to observe the effect of HMGA2 on the invasion and migration of miR-129 mimic CAOV cells. Results: The expression of miR-129 in HGSOC tissues and cell lines were significantly lower than that in normal fallopian tubes and normal ovarian epithelial cells(P<0.05). HMGA2 was significantly higher in HGSOC tissues than in normal fallopian tubes(P<0.05), negatively correlated with the expression of miR-129(r=-0.712, P<0.05). Compared with the control group, the invasion and migration ability of CAOV cells in miR-129 mimic group decreased, and the expression of HMGA2 mRNA and protein decreased(P<0.05);bioinformatics software prediction and luciferase reporter gene experiments confirmed that HMGA2 is a target gene of miR-129;compared with miR-129+ control group, miR-129 mimic+HMGA2 group has significantly increased invasion and migration ability(all P<0.05). Conclusion: Mi R-129 is lowly expressed in HGSOC tissues, and it can inhibit the invasion and migration of ovarian cancer cells by targeting down-regulation of HMGA2.