microRNA-145对结肠癌细胞株SW480侵袭能力的影响

Effect of miR-145 on invasion ability of Colonic Cancer cell line SW480.

目的 研究miR-145 对结肠癌细胞株SW480 侵袭能力以及对FSCN1 表达的影响,探讨其在结肠癌侵袭转移中的机制. 材料与方法 本实验采用化学方法合成成熟的miR-145 寡核苷酸模拟物（mimics）,以lipofectamine 2000 瞬时转染结肠癌细胞系SW480;转染后48小时,采用实时聚合酶反应（Real-time PCR）检测转染后各组miR-145 的表达;转染后24 小时,使用预铺基质胶的Transwell 模型检测细胞侵袭能力;转染后72 小时,通过蛋白免疫印记（Western Blot）检测FSCN1 蛋白表达. 结果 与对照组比较,miR-145 模拟物（miR-145 mimics）组细胞miR-145 表达明显升高（P〈0.05）;穿过Transwell 小室聚碳酯膜细胞数较对照组明显下降（P〈0.05）,蛋白印记（Western Blot）显示FSCN1 表达明显下降（P〈0.05）.结论 1、miR-145 可降低结肠癌细胞株SW480 的侵袭能力;FSCN1 为miR-145 的靶基因,miR-145 可能通过对其负调控进而抑制肿瘤细胞侵袭,证明其可能在细胞侵袭过程中起到重要作用.

Objective To investigate the effect of miR-145 on invasion ability of colorectal cancer cell line SW480 and regulation to FSCN1 ,meanwhile to study the mechanism.Material ＆ Methods Colonic cancer cell line SW480 was transient transfected with chemically synthesized maturemiR-145 mimic oligonucleotides by lipofecatamine 2000 .At 48h after transfection , the expression of miRNA-145 was identified by Real-time polymerasechain reaction （PCR） . At 24h after transfection , the pre-coated matrigel model was used for the invasion assay .At 72h after transfection , targeting proteinexpression was evaluated by Western Blot,.Results After transfection of miR-145 mimics ,as compared with control group, the miR-145was increased significantly in miR-145 mimics group, meanwhile the protein expression of FSCN1 was down-regulated （P〈0.05）,and the invasionability was down-regulated significantly（P〈0.05）.Conclusion One miR-145 suppress invasion ability of colonic cancer cell line SW480; Two：Evaluated as a target gene of miR-145, FSCN1 is negative regulated by miR-145 to suppress invasion ability of tuomor cells ,and may play an importantrole in metastatic process of tumor cells.