微小RNA-454-3p对子宫内膜癌细胞生物学行为的影响

Effect of microRNA-454-3p on biological behavior of endometrial cancer cells

目的研究微小RNA-454-3p(miR-454-3p)对子宫内膜癌(EC)细胞增殖、侵袭、迁移能力的影响及其机制。方法用miTED数据库分析miR-454-3p在各组织中的表达水平,用starBase数据库分析EC患者中高表达miR-454-3p组和低表达miR-454-3p组的生存率。用Targetscan预测miR-454-3p的下游靶基因。用Kaplan-Meier Plotter数据库分析EC患者中高表达检测磷酸酯酶与张力蛋白同源物(PTEN)组(高队列的平均表达水平为2412.45)和低表达PTEN组(低队列的平均表达水平为733.15)的生存率。将Ishikawa细胞分为NC-mimics组(转染NC-mimics)、mimics组(转染miR-454-3p mimics)、NC-inhibitor组(转染NC-inhibitor)、inhibitor组(转染miR-454-3p inhibitor)。用细胞计数试剂盒-8(CCK-8)实验检测细胞的增殖能力,用Transwell和划痕实验分别评估细胞侵袭和迁移能力,用实时定量聚合酶链反应(qRT-PCR)检测PTEN RNA的表达水平。结果EC患者肿瘤组织与正常组织相比,miR-454-3p的表达水平增高(1.57 vs 3.26,P<0.05),且低表达miR-454-3p组的生存率显著高于高表达miR-454-3p组(0.74 vs 0.42,P<0.05)。NC-mimics组、mimics组、NC-inhibitor组、inhibitor组细胞增殖能力分别为3.73±0.02、5.40±0.02、2.06±0.05和1.95±0.05,侵袭细胞数分别为(116±17)、(154±19)、(855±165)和(447±44)个,迁移细胞数分别为(116±8)、(154±27)、(1518±50)和(1132±175)个,划痕愈合率分别为(20.00±8.00)%、(39.00±2.00)%、(84.00±1.00)%和(52.00±1.00)%,PTEN mRNA表达水平分别为1.16±0.03、0.94±0.02、0.85±0.14和1.22±0.07。mimics组的上述指标与NC-mimics组相比,inhibitor组的上述指标与NC-inhibitor组相比,在统计学上差异均有统计学意义(均P<0.05)。EC患者肿瘤组织中PTEN的表达水平显著降低(33.45 vs 17.17,P<0.05),且低表达PTEN组患者生存率显著高于高表达PTEN组(41%vs 93%,P<0.05)。结论miR-454-3p可能通过负调控PTEN的表达促进EC细胞的增殖、侵袭和迁移能力。

Objective To investigate the effect of microRNA-454-3p(miR-454-3p)on the proliferation,invasion,and migration ability of endometrial cancer(EC)cells and its mechanism.Methods Analyze the expression levels of miR-454-3p in various tissues using the miTED database.Analyze the survival rates of high expression miR-454-3p and low expression miR-454-3p groups in EC patients using the starBase database.Use Targetscan to predict downstream target genes of miR-454-3p.Use the Kaplan Meier Plotter database to analyze the sur viv al rates of the high expression detection of phosphatase and tensin homolog(PTEN)group(average expression level in the high cohort:2412.45)and the low expression of PTEN(average expression level in the low cohort:733.15)in EC patients.Ishikawa cells were divided into NC-mimics group(transfected with NC-mimics),mimics group(transfected with miR-454-3p mimics),NC-inhibitor group(transfected with NC-inhibitor),and inhibitor group(transfected with miR-454-3p inhibitor).Detect the proliferation ability of cells using cell counting kit-8(CCK-8)assay;evaluate cell invasion and migration abilities using Transwell and scratch experiments,respectively;detection of PTEN RNA expression levels using real-time quantitative polymerase chain reaction(qRT-PCR).Results Compared with normal tissue,the expression level of miR-454-3p in tumor tissue of EC patients increased(1.57 vs 3.26,P<0.05),and the survival rate of EC patients with low expression of miR-454-3p increased(0.74 vs 0.42,P<0.05).The cell proliferation ability of the NC mimics group,mimics group,NC inhibitor group and inhibitor group were 3.73±0.02,5.40±0.02,2.06±0.05 and 1.95±0.05;the number of invading cells were 116±17,154±19,855±165 and 447±44;the number of migrating cells were 116±8,154±27,1518±50 and 1132±175;the scratch healing rates were(20.00±8.00)%、(39.00±2.00)%、(84.00±1.00)%and(52.00±1.00)%;the expression levels of PTEN mRNA were 1.16±0.03,0.94±0.02,0.85±0.14 and 1.22±0.07,respectively.The differences in the above indicators between the mimics group and the NC mimics group,as well as between the inhibitor group and the NC inhibitor group,were statistically significant(all P<0.05).The expression level of PTEN in tumor tissue of EC patients decreased(33.45 vs 17.17,P<0.05),and the survival rate of EC patients with low expression of PTEN increased(41%vs 93%,P<0.05).Conclusion miR-454-3p may promote the proliferation,invasion,and migration of EC cells by negatively regulating the expression of PTEN.