MiR-148a-3p通过靶向SRPK2抑制结肠癌细胞转移

目的:MicroRNA (miRNA/miR)参与结肠癌各种生物学过程。目前，miR-148a-3p在结肠癌中的作用还不完全清楚。本研究旨在探讨miR-148a-3p对结肠癌细胞转移及侵袭能力的影响及机制。方法:实时定量PCR （qRT-PCR）检测结肠癌细胞系中miR-148a-3p及SRPK2 mRNA的表达，Western blot检测SRPK2 蛋白的表达。使用miR-148a-3p mimic， miR-148a-3p inhibitor调节HCT-116及SW480细胞中miR-148a-3p的水平。通过划痕及transwell实验检测miR-148a-3p对结肠癌细胞迁移及侵袭能力的影响。生物信息学分析预测miR-148a-3p的靶点，荧光素酶报告实验验证。Western blot及qRT-PCR检测miR-148a-3p对结肠癌细胞上皮-间质转化及SRPK2表达的影响。结果:与正常结直肠粘膜细胞FHC相比，结肠癌细胞系中miR-148a-3p水平降低（P<0.05）。结肠癌细胞中miR-148a-3p过表达可以明显抑制结肠癌细胞转移及侵袭能力（P<0.05）。相反的，敲低miR-148a-3p结肠癌细胞转移及侵袭能力增强（P<0.05）。荧光素酶报告系统结果提示SRPK2是miR-148a-3p的直接作用靶点。过表达miR-148a-3p可以抑制SRPK2在结肠癌中的表达（P<0.05），但是敲低miR-148a-3p时SRPK2表达明显增高（P<0.05）。结论:MiR-148a-3p可能通过靶向SRPK2抑制结肠癌细胞的转移及侵袭能力。MiR-148a-3p可能成为诊断和治疗结肠癌的靶点之一。

MiR-148a-3p inhibits colon cancer cell metastasis by targeting SRPK2

Objective: To investigate the effects of miR-148a-3p on colon cancer cell metastasis and invasion. Methods: The expressions of miR-148a-3p and SRPK2 mRNA in colon cancer cell lines were detected by quantitative real-time PCR (qRT-PCR). The expression of SRPK2 protein was detected by Western blot. MiR-148a-3p mimic, miR-148a-3p inhibitor was used to regulate the level of miR-148a-3p in HCT-116 and SW480 cells. The effects of miR-148a-3p on the migration and invasion of colon cancer cells were examined by wound-healing and transwellassay. The target of miR-148a-3p was predicted by bioinformatics analysis and was verified by luciferase reporter assay. The effects of miR-148a-3p on epithelial-mesenchymal transition and SRPK2 expression in colon cancer cells were detected by Western blot and qRT-PCR. Results: MiR-148a-3p levels were reduced in CRC cell lines as compared to normal colon cell FHC (P<0.05). Overexpression of miR-148a-3p in colon cancer cells significantly inhibited the metastasis and invasion of colon cancer cells (P<0.05). In contrast, knockdown of miR-148a-3p showed an increased metastasis and invasion of colon cancer cells (P<0.05). SRPK2 is a direct target of miR-148a-3p. Overexpression of miR-148a-3p inhibited the expression of SRPK2 in colon cancer (P<0.05), while the expression of SRPK2 was significantly increased in miR-148a-3p CRC cells (P<0.05). Conclusion: MiR-148a-3p can inhibit the metastasis and invasion of colon cancer cells by targeting SRPK2. MiR-148a-3pmight be one of the targets for the diagnosis and treatment of colon cancer.