| 李莹,孙万仆,安志强,等.微小 RNA?671?5p对人结肠癌细胞增殖、迁移和侵袭的影响[J].安徽医药,2020,24(5):942-946. |
| 微小 RNA?671?5p对人结肠癌细胞增殖、迁移和侵袭的影响 |
| miR?671?5p inhibits colon cancer cell proliferation，migration and invasion by targeting CBX4 |
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| DOI：10.3969/j.issn.1009?6469.2020.05.024 |
| 中文关键词: 结肠肿瘤 微小 RNA?671?5p 染色盒同源物 4 细胞增殖 迁移 侵袭 酪氨酸激酶 1/信号转导子与转录激活子 3 |
| 英文关键词: Colonic neoplasms miR?671?5p Chromobox homolog 4 Cell proliferation Migration Invasion Ianus kinase 1/signal transducerand activator of transcription 3 |
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| 中文摘要: |
| 目的探讨 miR?671?5p对结肠癌细胞增殖、迁移和侵袭的影响及作用机制。方法培养人正常结肠上皮细胞 NCM460和结肠癌细胞 SW480、SW620、HT29和 LOVO，实时荧光定量 PCR(RT?qPCR)检测 miR?671?5p和染色盒同源物 4(CBX4)mRNA水平。转染 miR?671?5p mimics(miR?671?5p组)、阴性对照( miR?NC组)至 SW480细胞，四甲基噻唑蓝染色法( MTT)检测细胞增殖，克隆实验检测细胞克隆形成能力， Transwell检测细胞迁移和侵袭，蛋白质印迹法( Western Blot)检测 CBX4、细胞周期蛋白D1(CyclinD1)、 p21、基质金属蛋白酶 9(MMP?9)、基质金属蛋白酶 14(MMP?14)及酪氨酸激酶 1/信号转导子与转录激活子 3(JAK1/STAT3)信号通路相关蛋白表达水平。双荧光素酶报告基因实验验证 miR?671?5p和 CBX4之间靶向关系。结果与 NCM460细胞比，结肠癌细胞 SW480、SW620、HT29和 LOVO中 miR?671?5p水平显著降低( P＜0.05)，CBX4 mRNA水平显著升高( P＜0.05)。与 miR?NC组比， miR?671?5p组 SW480细胞培养 24、48、72 h后的 OD值、克隆形成率、迁移和侵袭细胞数及 CyclinD1、MMP?9、MMP?14、p?JAK1和 p?STAT3蛋白水平显著降低( P＜0.05)，p21蛋白水平显著升高( P＜0.05)。 miR?671?5p负调控 CBX4表达，过表达 CBX4降低了 miR?671?5p过表达对 SW480细胞增殖、迁移和侵袭及 JAK1/STAT3信号通路的影响。结论 miR?671?5p过表达可抑制结肠癌细胞的增殖、迁移和侵袭，其作用机制与下调 CBX4表达、抑制 JAK1/STAT3信号通路的激活有关。 |
| 英文摘要: |
| Objective To investigate the effects of miR?671?5p on proliferation，migration and invasion of colon cancer cells and the mechanisms.Methods Human normal colonic epithelial cells NCM460 and colon cancer cells SW480，SW620，HT29 and LO?VO were cultured.The miR?671?5p and CBX4 mRNA levels were detected by RT?qPCR.The miR?671?5p mimics(miR?671?5p group)and the negative control(miR?NC group)were transfected into SW480 cells.MTT assays was used to detect cell prolifera?tion，cloning assay was detected cell clone formation ability，Transwell was used to detect cell migration and invasion and Western Blot assay was used to detect the levels of CBX4，CyclinD1，p21，MMP?9，MMP?14 protein and JAK1/STAT3 signaling pathway?re?lated protein.The dual luciferase reporter gene assay validated the targeting relationship between miR?671?5p and CBX4.Results Compared with the NCM460 cells，the levels of miR?671?5p in colon cancer SW480，SW620，HT29 and LOVO were significantly decreased(P＜0.05)，and CBX4 mRNA levels were significantly increased(P＜0.05).Compared with the miR?NC group，the OD value，colony formation rate，migration and invasion cell number and the levels of CyclinD1，MMP?9，MMP?14，p?JAK1 and p?STAT3 in the miR?671?5p group were significantly decreased(P＜0.05)，and the level of p21 protein was significantly increased(P＜0.05).miR?671?5p negatively regulated CBX4 expression，overexpression of CBX4 reduced the effect of miR?671?5p overex? pression on proliferation，migration and invasion of SW480 cells and JAK1/STAT3 signaling pathway.Conclusion Overexpressionof miR?671?5p can inhibit the proliferation，migration and invasion of colon cancer cells，which is related to down?regulation of CBX4 expression and inhibition of JAK1/STAT3 signaling pathway activation. |
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