miRNA-126调控Wnt/β-catenin信号通路对子宫平滑肌肉瘤生物学功能的影响

Effect of miRNA-126 Regulation of Wnt/β-catenin Signaling Pathway on the Biological Function of Uterine Leiomyosarcoma

目的研究miRNA-126在子宫平滑肌肉瘤(ULMS)组织中的表达水平及临床意义,探讨其通过下调Wnt/β-catenin信号通路抑制ULMS细胞增殖和转移能力的作用机制。方法采用qRT-PCR检测miRNA-126在ULMS组织和子宫平滑肌瘤组织中的表达水平,并分析miRNA-126表达与ULMS患者临床病理参数、复发转移及生存率的关系。常规培养ULMS细胞SK-UT-1,随机分为NC对照组和miRNA-126 mimics实验组,分别转染NC mimics和miRNA-126 mimics,qRT-PCR检测各组细胞中miRNA-126的表达;CCK8实验检测各组细胞的增殖能力;Transwell实验检测各组细胞的转移能力;TOP/FOP Flash实验检测各组细胞Wnt/β-catenin信号通路活性;western blotting检测各组细胞中Wnt/β-catenin信号通路关键蛋白β-catenin在细胞核中的表达水平,及其相关蛋白Slug、Snail和CyclinD1的表达。结果qRT-PCR检测结果显示,与子宫平滑肌瘤组织相比,miRNA-126在ULMS组织中的表达显著降低(P<0.05),且miRNA-126在肿瘤直径大和存在转移的ULMS患者中表达显著减少(P<0.05),miRNA-126表达对ULMS患者复发转移和生存率无明显影响(P>0.05)。与NC组相比,miRNA-126 mimics组SK-UT-1细胞中的miRNA-126表达水平显著增加(P<0.05),SK-UT-1细胞的增殖和转移能力显著降低(P<0.05),Wnt/β-catenin信号通路关键蛋白β-catenin及相关蛋白Slug、Snail和CyclinD1表达均显著降低(P<0.05)。结论miRNA-126在ULMS组织中表达降低,miRNA-126可能通过下调Wnt/β-catenin信号通路抑制ULMS细胞的增殖和转移。

Objective To investigate the expression level and clinical significance of miRNA-126 in ULMS tissues,and its mechanism of inhibiting the proliferation and metastasis of uterine Leiomyosarcoma(ULMS)cells by downregulating Wnt/β-catenin signaling pathway.Methods The expression levels of miRNA-126 in ULMS and uterine leiomyoma tissues were detected by qRT-PCR,and miRNA-126 expression was analyzed in relation to clinicopathological parameters,recurrent metastasis,and survival rate of ULMS patients.ULMS cells SK-UT-1 were routinely cultured and randomly divided into NC control group and miRNA-126 mimics experimental group,transfected with NC mimics and miRNA-126 mimics,respectively.qRT-PCR was performed to detect the expression of miRNA-126 in the cells of each group;CCK8 assay was performed to detect the proliferation ability of cells in each group;the metastatic ability of cells was detected with Transwell assay in each group;the Wnt/β-catenin signaling pathway activity of cells was detected with TOP/FOP Flash in each group;Western blotting was conducted to detect the expression level ofβ-catenin,a key protein of the Wnt/β-catenin signaling pathway,in the nucleus,and its related proteins Slug,Snail and CyclinD1 in the cells of each group.Results The results of qRT-PCR showed that compared with uterine leiomyoma tissue,the expression of miRNA-126 was significantly reduced in ULMS tissue(P<0.05),and miRNA-126 expression was noticeably reduced in ULMS patients with large tumor diameter and the presence of metastasis(P<0.05).miRNA-126 expression had no significant effect on recurrent metastasis and survival in ULMS patients(P>0.05).Compared with the NC group,the miRNA-126 expression level in SK-UT-1 cells in the miRNA-126 mimics group was significantly increased(P<0.05),the proliferation and metastasis of SK-UT-1 cells were significantly reduced(P<0.05),and the key protein of the Wnt/β-catenin signaling pathway,β-catenin,and the related proteins Slug,Snail and CyclinD1 expression were significantly reduced(P<0.05).Conclusion miRNA-126 expression was reduced in ULMS tissues and miRNA-126 may inhibit the proliferation and metastasis of ULMS cells by down-regulating the Wnt/β-catenin signaling pathway.