Department of Digestive Endoscopy, Second Affiliated Hospital of Jilin University, Changchun, China.
Department of Digestive Endoscopy, Second Affiliated Hospital of Jilin University, Changchun, China.
Chem Biol Interact. 2018 Jun 1;289:9-14. doi: 10.1016/j.cbi.2018.04.016. Epub 2018 Apr 13.
MicroRNAs (miRNAs) can function as tumor suppressor or oncogenic genes. The putative targets of miR-223 include tumor suppressor gene, RhoB. Here we sought to investigate the role of miR-223-RhoB signaling pathway in proliferation of colon cancer. We used Western blot, immunofluorescence staining, or RT-PCR to detect expression levels of miR-223 and RhoB in colon adenocarcinoma and adjacent non-cancerous tissue samples, or in human colon adenocarcinoma cell lines. MTT assay was used to determine proliferation and apoptosis in cell lines. We further used Western blot to determine levels of cell cycle regulators CDK1 and Cyclin B1 with anti-miR-223 or apoptosis with overexpression of RhoB. The expression level of miR-223 was significantly upregulated in clinical samples and cell lines of colon adenocarcinoma, in contrast to down-regulation of RhoB. In addition, we showed that inhibition of miR-223 led to upregulation of RhoB and in turn suppression of proliferation of colon adenocarcinoma. Moreover, inhibition of miR-223 or overexpression of RhoB induced cell arrest or apoptosis in colon adenocarcinoma. These results suggest that miR-223-RhoB signaling pathway plays an important role in modulation of proliferation, cell arrest, and apoptosis in colon cancer.
微小 RNA（miRNAs）可以作为肿瘤抑制基因或癌基因发挥作用。miR-223 的假定靶基因包括肿瘤抑制基因 RhoB。本研究旨在探讨 miR-223-RhoB 信号通路在结肠癌增殖中的作用。我们采用 Western blot、免疫荧光染色或 RT-PCR 检测结肠癌及癌旁正常组织样本或人结肠腺癌细胞系中 miR-223 和 RhoB 的表达水平。MTT 法检测细胞系的增殖和凋亡。我们进一步采用 Western blot 检测细胞周期调节因子 CDK1 和 Cyclin B1 的水平（用抗 miR-223 处理）或用 RhoB 过表达检测凋亡。miR-223 在结肠癌临床样本和细胞系中的表达水平明显上调，而 RhoB 的表达水平则下调。此外，我们发现抑制 miR-223 可导致 RhoB 上调，从而抑制结肠腺癌细胞的增殖。此外，抑制 miR-223 或过表达 RhoB 可诱导结肠腺癌细胞停滞或凋亡。这些结果表明，miR-223-RhoB 信号通路在调节结肠癌的增殖、细胞停滞和凋亡中发挥重要作用。
