a Department of Gastroenterology, Zhujiang Hospital , Southern Medical University , Guangzhou , China.
b Department of Radiation Oncology , Tianjin Medical University Cancer Institute and Hospital , Tianjin , China.
Artif Cells Nanomed Biotechnol. 2019 Dec;47(1):1722-1729. doi: 10.1080/21691401.2019.1606006.
Mir-10b has been reported as a key regulator of metastasis in many human tumours. Moreover, it has also been regarded as a prognostic marker and therapeutic target of colorectal cancer (CRC). Whether miR-10b could affect the metastasis and proliferation of CRC is unclear. MiR-10b expression was detected by qPCR in human CRC tissues and cell line, Luciferase activity was employed for miR-10b binding to the 3`UTR of KLF4, Genes expression were examined by western blot, and mRNA by qPCR. PI and Annexin V staining were used to evaluate the cell cycle and apoptosis. Cell proliferation was detected with MTT, and cell migration and invasion were performed with Transwell assay. We found that miR-10b expression was up-regulated in metastatic CRC tissues and cell lines. Inhibition of miR-10b prevented cancer cell metastasis and growth by inducing cell-cycle arrest and apoptosis in vitro. Moreover, we found that KLF4 was a direct target of miR-10b. MiR-10b inhibitor led to the up-regulation of E-cadherin expression and the down-regulation of cyclin D1, which were partly abrogated after silencing KLF4.
miR-10b 已被报道为许多人类肿瘤转移的关键调节因子。此外,它也被认为是结直肠癌(CRC)的预后标志物和治疗靶点。miR-10b 是否能影响 CRC 的转移和增殖尚不清楚。通过 qPCR 检测人 CRC 组织和细胞系中的 miR-10b 表达,通过荧光素酶活性检测 miR-10b 与 KLF4 的 3'UTR 的结合,通过 Western blot 检测基因表达,通过 qPCR 检测 mRNA。PI 和 Annexin V 染色用于评估细胞周期和凋亡。通过 MTT 检测细胞增殖,通过 Transwell 测定检测细胞迁移和侵袭。我们发现 miR-10b 在转移性 CRC 组织和细胞系中表达上调。抑制 miR-10b 可通过诱导细胞周期停滞和凋亡来阻止体外癌细胞转移和生长。此外,我们发现 KLF4 是 miR-10b 的直接靶标。miR-10b 抑制剂导致 E-钙粘蛋白表达上调和细胞周期蛋白 D1 表达下调,沉默 KLF4 后部分逆转。