Guizhou Provincial Key Laboratory of Pathogenesis and Drug Research on Common Chronic Diseases, Guizhou Medical University, Guiyang, Guizhou 550009, P.R. China.
Department of Biliary‑Pancreatic Surgery, Affiliated Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430060, P.R. China.
Int J Mol Med. 2020 Mar;45(3):886-896. doi: 10.3892/ijmm.2020.4474. Epub 2020 Jan 23.
It is widely known that abnormal regulation of microRNAs (miRNAs/miRs) may contribute to the occurrence or development of tumors. The objective of the present study was to elucidate the function and underlying mechanism of miR‑137 in the progression of cholangiocarcinoma (CCA). The expression levels of miR‑137 in CCA tissues and cell lines were measured using reverse transcription‑quantitative PCR. The role of miR‑137 in the proliferation of CCA cells was assessed using the Cell Counting Kit‑8 assay, colony formation assay and cell cycle distribution analysis, while its effects on the migration and invasion of CCA cells were evaluated using Transwell assays. The function of miR‑137 on CCA growth in vivo was also investigated using a xenograft mouse model. Furthermore, the association between miR‑137 and Wnt family member 2B (WNT2B) was analyzed using bioinformatics, double luciferase assay and western blotting. It was verified that the expression of miR‑137 was low in CCA tissues and cell lines, whereas increased expression of miR‑137 significantly suppressed cell proliferation, decreased colony formation ability and induced G1 phase arrest. miR‑137 overexpression suppressed the migration and invasion ability of TFK‑1 and HuCCT1 cells. Furthermore, the results of the xenograft mouse model assays revealed that miR‑137 overexpression decreased tumor growth in vivo. The results of bioinformatics analysis and dual luciferase reporter assays demonstrated that WNT2B is directly regulated by miR‑137. The expression of WNT2B and Wnt‑pathway‑related proteins was decreased when miR‑137 was overexpressed. Restoring the expression of WNT2B notably reversed the inhibitory effect of miR‑137 on CCA cells. Therefore, the findings of the present study demonstrated that miR‑137 acts as a suppressor in CCA and inhibits CCA cell proliferation, migration and invasion through suppressing the expression of WNT2B.
众所周知，微小 RNA（miRNAs/miRs）的异常调节可能导致肿瘤的发生或发展。本研究旨在阐明 miR-137 在胆管癌（CCA）进展中的作用及其潜在机制。采用逆转录定量 PCR 检测 CCA 组织和细胞系中 miR-137 的表达水平。采用细胞计数试剂盒-8 检测、集落形成检测和细胞周期分布分析评估 miR-137 对 CCA 细胞增殖的作用，采用 Transwell 检测评估 miR-137 对 CCA 细胞迁移和侵袭的影响。还通过异种移植小鼠模型研究了 miR-137 对 CCA 生长的体内作用。此外，还通过生物信息学、双荧光素酶报告基因检测和 Western blot 分析分析了 miR-137 与 Wnt 家族成员 2B（WNT2B）之间的关联。验证结果显示，miR-137 在 CCA 组织和细胞系中的表达水平较低，而 miR-137 表达增加则显著抑制细胞增殖、降低集落形成能力并诱导 G1 期阻滞。miR-137 过表达抑制 TFK-1 和 HuCCT1 细胞的迁移和侵袭能力。此外，异种移植小鼠模型试验结果表明，miR-137 过表达可减少体内肿瘤生长。生物信息学分析和双荧光素酶报告基因检测结果表明，WNT2B 受 miR-137 直接调控。miR-137 过表达时，WNT2B 及其 Wnt 通路相关蛋白的表达降低。恢复 WNT2B 的表达显著逆转了 miR-137 对 CCA 细胞的抑制作用。综上所述，本研究结果表明，miR-137 在 CCA 中作为一种抑制因子发挥作用，通过抑制 WNT2B 的表达来抑制 CCA 细胞的增殖、迁移和侵袭。
