过表达miR-145-5p通过下调IGF1R抑制食管鳞状细胞TE-10细胞的恶性生物学行为

目的:探究miR-145-5p对食管鳞状细胞癌TE-10细胞增殖、侵袭、迁移和上皮间质转化(epithelial-mesenchymal transition,EMT)等恶性生物学行为的分子机制。方法:q PCR法检测miR-145-5p在食管鳞状细胞癌细胞和正常食管上皮细胞中的表达水平。采用双荧光素酶报告基因实验检测miR-145-5p与胰岛素样生长因子1受体(insulin-like growth factor 1 receptor,IGF1R)的靶向调控关系,Western blotting检测IGF1R蛋白和EMT相关蛋白的表达,CCK-8法和Transwell检测miR-145-5p/IGF1R分子轴对TE-10细胞增殖、侵袭、迁移的影响。结果:miR-145-5p在3株食管鳞状细胞癌细胞中低表达且在TE-10细胞中表达最低(P<0.01或P<0.05)。过表达miR-145-5p可显著抑制TE-10细胞的增殖、侵袭、迁移和EMT进程(P<0.01或P<0.05)。双荧光素酶报告基因证实miR-145-5p靶向下调IGF1R表达(P<0.01)。回复实验进一步证实,与单独过表达IGF1R相比,同时过表达miR-145-5p和IGF1R能显著缓解IGF1R对TE-10细胞的增殖、侵袭、迁移和EMT进程的促进作用(P<0.01或P<0.05)。结论:过表达miR-145-5p通过靶向下调IGF1R进而抑制了食管鳞状细胞癌TE-10细胞的增殖、侵袭、迁移和EMT进程。

Over-expression of miR-145-5p inhibits malignant biological behaviors of esophageal squamous cells carcinoma via down-regulating IGF1R

Objective: To explore the mechanism of miR-145-5p on malignant biological behaviors, such as pro-liferation, invasion,migration and epithelial-mesenchymal transition (EMT), of esophageal squamous cell carcinoma (ESCC) TE-10 cells. Methods: The expression of miR-145-5p in ESCC cell lines and normal cells was detected by PCR. Dual luciferase reporter gene assay was used to detect the targeted regulation between miR-145-5p and insulin-like growth factor 1 receptor (IGF1R). The expres-sions of IGF1R protein and EMT related proteins were detected by Western blotting. Transwell assay and CCK-8 assay were carried out to detect the effects of miR-145-5p/IGF1R axis on the proliferation, migration and invasion of TE-10 cells. Results: miR-145-5p was down-regulated in ESCC cell lines with the lowest expression in TE-10 cells (P<0.01 or P<0.05). Over-expression of miR-145-5p significantly inhibited proliferation, invasion, migration and EMT of TE-10 cells (P<0.01 or P<0.05). Dual luciferase reporter gene assay con-firmed that miR-145-5p targetedly down-regulated IGF1R expression (P<0.01). The restora-tion experiments further confirmed that simultaneous over-expression of miR-145-5p and IGF1R significantly attenuated the promotion effect of IGF1R on proliferation, invasion, migration and EMT of TE-10 cells (P<0.01 or P<0.05). Conclusions: Over-expression of miR-145-5p inhibits proliferation, invasion, migration and EMT of ESCC TE-10 cells by down-regulating IGF1R.