上调miRNA-22表达对卵巢癌细胞株SKOV-3侵袭的影响及与VEGF和P53蛋白表达的关系研究

Effect of up-regulation of microRNA-22 expression on invasion of ovarian cancer cell line SKOV-3 and its relationship with expression of vascular endothelial growth factor and P53 protein

目的研究上调miRNA-22表达对卵巢癌细胞株SKOV-3侵袭的影响及与血管内皮生长因子(VEGF)和P53蛋白表达的关系。方法前瞻性收集新疆维吾尔自治区人民医院的卵巢癌组织标本112例(A组)、卵巢癌旁组织标本85例(B组)及正常卵巢组织标本75例(C组)。采用实时荧光定量PCR法对三组标本中的miRNA-22表达量进行检测。将卵巢癌SKOV-3细胞株分为转染miRNA-22-mimic(转染组)、miRNA-22-NC(空白载体组)及正常对照组。采用实时荧光定量PCR法对各组细胞中的miRNA-22表达量进行测定,通过Transwell小室法对各组细胞的侵袭能力进行检测,并用Western blot法对各组细胞中的P53及VEGF蛋白表达情况进行检测。结果A组miRNA-22表达量较B、C组均明显下降(P<0.05),而B、C组miRNA-22表达量的比较,并无显著差异(P>0.05)。相比正常对照组与空白载体组,转染组细胞中的miRNA-22表达量明显升高(P<0.05),而正常对照组与空白载体组细胞中miRNA-22表达量的比较,无显著差异(P>0.05)。Transwell小室法检测结果发现,转染组卵巢癌SKOV-3细胞株侵袭能力较正常对照组与空白载体组明显减弱(P<0.05),而正常对照组与空白载体组侵袭细胞数的比较,无显著差异(P>0.05)。Western blot法检测结果发现,转染组细胞中P53和VEGF蛋白表达水平较正常对照组与空白载体组明显下降(P<0.05),而正常对照组与空白载体组细胞中P53和VEGF蛋白表达水平的比较,无显著差异(P>0.05)。结论上调miRNA-22表达可起到阻滞卵巢癌SKOV-3细胞株侵袭的作用,其机制可能与P53和VEGF蛋白表达水平下降密切相关。

Objective To study the effect of up-regulation of the expression of microRNA-22 on the invasion of ovarian cancer cell line SKOV-3 and its relationship with the expression of vascular endothelial growth factor(VEGF)and P53protein.Methods 112 cases of ovarian cancerspecimens(group A),85 cases of adjacent ovarian cancerspecimens(group B)and 75 cases of normal ovarian tissuespecimens(group C)were collected,and the expression of microRNA22 in the three groups was detected by real-time fluorescence quantitative PCR.SKOV-3 ovarian cancer cell lines were divided into transfected microRNA-22-mimic(transfection group),microRNA-22-NC(blank vector group)and normal control group.Real-time fluorescence quantitative PCR was used to detect the expression of microRNA-22 in each group.Transwell chamber method was used to detect the invasive ability of each group,and Western blot method was used to detect the expression of P53 and VEGF protein in each group of cells.Results The expression of microRNA-22 in group A was significantly decreased compared with group B and C(P<0.05),but there was no significant difference in the expression of microRNA-22 between group B and C(P>0.05).Compared with the normal control group and the blank vector group,the expression of microRNA-22 in the transfected group increased significantly(P<0.05),while there was no significant difference between the normal control group and the blank vector group(P>0.05).Transwell cell assay showed that the invasive ability of SKOV-3 cell line in transfected group was significantly weaker than that in normal control group and blank vector group(P<0.05),while there was no significant difference in the number of invasive cells between normal control group and blank vector group(P>0.05).Western blot assay showed that the expression of P53 and VEGF in the transfected cells was significantly decreased compared with the normal control group and the blank vector group(P<0.05).There was no significant difference between the normal control group and the blank vector group(P>0.05).Conclusion Upregulation of the expression of microRNA-22 can block the invasion of SKOV-3 cell line in ovarian cancer,and its mechanism may be closely related to the decrease of the expression of P53 and VEGF.