Background

To investigate the effect of miR-188-5p overexpression on the invasion and migration of cultured lung cancer cells, and on related cellular mechanisms that underlie epithelial mesenchymal transition (EMT).

Methods

Human lung cancer cell line 95D was transfected with miR-188-5p mimic. Quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot were performed to quantify the expression levels of genes including E-cadherin, Snail, α-SMA, and MGAT3. Changes in cell motility, invasion and proliferation were studied using scratch migration assay, transwell invasion assay, and colony formation assay, respectively. The expression levels of EMT-related proteins and MGAT3 protein were also determined via immunofluorescent staining. The ability of miR-188-5p to regulate its target gene, MGAT3, was assessed using dual luciferase activity assay.

Results

Lung cancer cell line 95D showed the lowest miR-188-5p expression level thus was used in this study. Transfection with miR-188-5p mimic significantly suppressed migration, invasion and clonal formation potency of 95D cells. Dual luciferase activity assay implicated that miR-188-5p exerts its negative regulatory effect on MGAT3 expression through recognizing the 3′ untranslated region (3′UTR) of the MGAT3 gene. Over-expression of miR-188-5p in 95D cells also remarkably increased E-cadherin protein expression and decreased the expression levels of Snail and α-SMA, which suppressed the EMT process.

Conclusion

MiR-188-5p reduces the expression of MGAT3 and inhibits the metastatic properties of a highly invasive lung cancer cell line, probably via targeted regulation of EMT process. Further research to explore the potential therapeutic value of miR-188-5p, both as a biomarker and as a drug candidate for the management of metastatic lung cancer may be warranted.

中文翻译：

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miR-188-5p 抑制 MGAT3 表达和肺癌细胞的上皮间质转化

背景

研究miR-188-5p过表达对培养的肺癌细胞侵袭和迁移的影响，以及上皮间质转化 (EMT) 的相关细胞机制。

方法

用miR-188-5p模拟物转染人肺癌细胞系 95D 。进行定量实时聚合酶链反应 (qRT-PCR) 和蛋白质印迹以量化包括E-cadherin、Snail、α-SMA和MGAT3在内的基因的表达水平。分别使用划痕迁移试验、transwell 侵袭试验和集落形成试验研究了细胞运动、侵袭和增殖的变化。还通过免疫荧光染色测定了 EMT 相关蛋白和 MGAT3 蛋白的表达水平。使用双荧光素酶活性测定评估miR-188-5p调节其靶基因MGAT3的能力。

结果

肺癌细胞系 95D 显示出最低的miR-188-5p表达水平，因此用于本研究。转染miR-188-5p模拟物显着抑制了 95D 细胞的迁移、侵袭和克隆形成效力。双荧光素酶活性测定表明miR-188-5p通过识别MGAT3基因的 3' 非翻译区 (3'UTR)对MGAT3表达发挥负调节作用。miR-188-5p在 95D 细胞中的过表达也显着增加了 E-cadherin 蛋白的表达，降低了 Snail 和 α-SMA 的表达水平，从而抑制了 EMT 过程。

结论

MiR-188-5p可能通过靶向调节 EMT 过程，降低 MCAT3 的表达并抑制高侵袭性肺癌细胞系的转移特性。可能需要进一步研究以探索miR-188-5p作为生物标志物和作为治疗转移性肺癌的候选药物的潜在治疗价值。