circ_0000885通过靶向miR-944调控胃癌细胞的增殖和凋亡

Circ_0000885 regulates proliferation and apoptosis of gastric cancer cells by targeting miR-944

  • 摘要:
    目的 探讨环状RNA _0000885(circ_0000885)对胃癌AGS细胞增殖和凋亡的影响及其分子机制。
    方法 选取30例胃癌组织及相应癌旁组织; 体外培养人正常胃黏膜细胞GES-1和胃癌细胞系AGS、HGC-27、N87、SNU-484。将AGS细胞分为NC组、si-NC组、si-circ_0000885组、微小RNA(miR)-NC组、miR-944组、si-circ_0000885+anti-miR-NC组、si-circ_0000885+anti-miR-944组。采用逆转录-实时定量聚合酶链反应(RT-qPCR)检测circ_0000885和miR-944的表达水平; 分别利用MTT、流式细胞术和Western blot检测细胞活性、凋亡和相关蛋白的表达; 双荧光素酶报告基因实验检测circ_0000885和miR-944的靶向关系。
    结果 与癌旁组织比较,胃癌组织中circ_0000885表达升高, miR-944表达降低,差异有统计学意义(P < 0.05)。与GES-1细胞比较,胃癌细胞系AGS、HGC-27、N87、SNU-484中的circ_0000885表达水平升高,miR-944表达水平降低,差异有统计学意义(P < 0.05)。敲减circ_0000885或过表达miR-944均降低了AGS细胞OD值和增殖细胞核抗原(PCNA)、Bcl-2蛋白水平,升高了细胞凋亡率和Bax蛋白水平(P < 0.05)。circ_0000885靶向调控miR-944表达,且下调miR-944逆转了敲减circ_0000885对AGS细胞增殖和凋亡的影响(P < 0.05)。
    结论 敲减circ_0000885可通过靶向上调miR-944抑制AGS细胞增殖,并促进凋亡。

     

    Abstract:
    Objective To investigate the effects of circRNA_0000885 (circ_0000885) on the proliferation and apoptosis of AGS cells in gastric cancer and its molecular mechanism.
    Methods The tissues of 30 cases of gastric cancer and corresponding adjacent tissues were selected. Human normal gastric mucosa cells GES-1 and gastric cancer cell lines AGS, HGC-27, N87, SNU-484 were cultured in vitro. AGS cells were divided into NC group, si-NC group, si-circ_0000885 group, MicroRNA(miR)-NC group, miR-944 group, si-circ_0000885+anti-miR-NC group and si-circ_0000885+anti-miR-944 group. The expression levels of circ_0000885 and miR-944 were detected by reverse transcriptionreal-time quantitative polymerasechain reaction (RT-qPCR); the cell activity, apoptosis and expression of related proteins were detected by MTT, flow cytometry and Western blot, respectively; the dual luciferase reporter gene assay was used to detect the targeting relationship between circ_0000885 and miR-944.
    Results Compared with adjacent tissues, the expression of circ_0000885 in gastric cancer tissues was significantly increased, and the expression of miR-944 was significantly decreased (P < 0.05). Compared with GES-1 cells, the expression of circ_0000885 in gastric cancer cell lines AGS, HGC-27, N87, SNU-484 was significantly increased, and the expression of miR-944 was significantly decreased (P < 0.05). Knockdown circ_0000885 or overexpression of miR-944 decreased OD value, proliferated cell nuclear antigen (PCNA) and Bcl-2 protein levels, and increased apoptosis rate and Bax protein levels in AGS cells (P < 0.05). The circ_0000885 targeted miR-944 expression, and down-regulation of miR-944 reversed the effects of circ_0000885 knockdown on proliferation and apoptosis of AGS cells (P < 0.05).
    Conclusion Knockdown of circ_0000885 inhibits the proliferation of AGS cells and promotes apoptosis by targeting up-regulation of miR-944.

     

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