miR-19 regulates PI3K/Akt signaling pathway by targeting PTEN to promote invasion and migration of endometrial cancer KLE cells / 中国肿瘤生物治疗杂志

@#[摘 要] 目的：分析miR-19对PTEN及PI3K-Akt通路的调节作用，揭示miR-19和PTEN对子宫内膜癌KLE细胞侵袭、迁移的影响及其机制。方法：收集2017年5月至2020年8月河北医科大学第一医院收治的74例子宫内膜癌患者经手术切除（术前未接受放化疗等治疗）且病理确诊为子宫内膜癌的肿瘤组织及对应癌旁组织，采用qPCR和WB法检测PTEN在子宫内膜癌组织中的表达水平以及转染miR-19模拟物或抑制物对KLE细胞中PTEN表达的影响。通过TargetScan及双荧光素酶报告基因实验预测并验证PTEN是否为miR-19的靶基因，将KLE细胞随机分为对照（NC）组、miR-19模拟物组和miR-19+PTEN组，分别转染阴性对照片段（miR-NC）、miR-19模拟物或共转染miR-19模拟物+PTEN过表达载体，采用Transwell和细胞划痕实验检测miR-19和PTEN对KLE细胞迁移和侵袭能力的影响，WB法检测对PI3K-Akt通路相关蛋白表达的影响。结果：子宫内膜癌组织中PTEN的mRNA和蛋白表达水平均明显低于癌旁组织（均P<0.01）。miR-19能与PTEN mRNA的3’-UTR特异性结合，上调miR-19的表达能抑制PTEN的mRNA和蛋白的表达，下调miR-19的表达则出现相反的结果（均P<0.01）。与miR-NC组相比，miR-19 模拟物组的迁移、侵袭细胞数以及划痕愈合率均显著升高（均P<0.01），而miR-19+PTEN组的细胞迁移和侵袭能力以及划痕愈合率较miR-19摸拟物组则明显降低（均P<0.01）；与miR-NC组相比，miR-19模拟物组中PTEN蛋白的表达明显降低，而p-Akt 308和p-Akt 473蛋白的表达明显升高，Akt蛋白的表达无明显变化（均P<0.01）；而在miR-19+PTEN组中，p-Akt 308和p-Akt 473蛋白的表达均明显低于miR-19模拟物组（均P<0.01）。结论：miR-19可能通过抑制PTEN的表达从而活化PI3K-Akt通路，进而促进子宫内膜癌KLE细胞的迁移和侵袭。

MiR-195 inhibits migration, invasion and epithelial-mesenchymal transition (EMT) of endometrial carcinoma cells by targeting SOX4

MicroRNAs (miRNAs) have been identified as potential biomarkers for endometrial carcinoma (EC) diagnosis, prognosisand therapy. The purpose of the present study was to investigate the detailed role and molecular mechanism of miR-195 inEC metastasis. qRT-PCR assay was performed to assess the expression of miR-195 and SRY-related high-mobility groupbox 4 (SOX4) mRNA in EC tissues and cells. The levels of N-cadherin, Vimentin, E-cadherin and SOX4 protein weredetermined by western blot. SOX4 protein expression in EC tissues was also determined by Immunohistochemistry (IHC)experiment. Transwell assay was used to analyze cell migration and invasion abilities. Dual-luciferase reporter assay andRNA Immunoprecipitation (RIP) assay were performed to confirm the targeted interaction between miR-195 and SOX4.Our data supported that miR-195 was downregulated and SOX4 was upregulated in EC tissues and cell lines. Upregulationof miR-195 inhibited migration, invasion and epithelial-mesenchymal transition (EMT) of EC cells. Moreover, SOX4 was adirect target of miR-195. MiR-195 overexpression-mediated anti-migration, anti-invasion and anti-EMT effects wereantagonized by SOX4 restoration in EC cells. In conclusion, our study suggested that miR-195 inhibited the migration,invasion and epithelial mesenchymal transition (EMT) of EC cells at least partly by targeting SOX4. Our study provided anovel underlying mechanism for EC metastasis and a promising therapeutic target for EC management.