抑制COL6A1基因表达对膀胱癌T24细胞增殖和侵袭能力的影响

杨荣权; 廖泽明; 蔡勇; 王超

doi:10.13201/j.issn.1001-1420.2019.06.009

抑制COL6A1基因表达对膀胱癌T24细胞增殖和侵袭能力的影响

- 荆门市第一人民医院泌尿外科(湖北荆门, 448000)

详细信息

通讯作者: 杨荣权,E-mail:yrq120@sina.cn

中图分类号: R737.14

收稿日期: 2017-12-18

Effect of down-regulated COL6A1 on proliferation and invasion of bladder carcinoma T24 cells

- Department of Urology, First People's Hospital of Jingmen, Jingmen, Hubei, 448000, China

More Information

Corresponding author: YANG Rongquan, E-mail: yrq120@sina.cn

Received Date: 18 December 2017

摘要

摘要: 目的:研究小干扰RNA抑制COL6A1基因表达对膀胱癌T24细胞增殖和侵袭能力的影响。方法:设计合成靶向COL6A1基因的siRNA,采用阳离子脂质体试剂瞬时转染膀胱癌细胞株T24,利用实时荧光定量PCR和Western blot检测RNA干扰后COL6A1基因的沉默效果,评价siRNA设计的合理性及RNA干扰抑制COL6A1表达的有效性,分别在RNA干扰后第1、2、3天应用噻唑蓝(MTT)比色法检测细胞增殖状态,并绘制生长曲线,应用Transwell法检测各组膀胱癌侵袭能力。结果:靶向COL6A1的siRNA成功抑制了膀胱癌细胞株T24中COL6A1 mRNA及蛋白表达,COL6A1-siRNA组蛋白相对表达量为阴性对照组的14.1%,mRNA相对表达量为空白对照组的20.5%;MTT比色法显示COL6A1-siRNA组细胞增殖能力显著减弱(P＜0.05),在第1、2、3天后重复检测,分别达到了空白对照组的75.6%、58.2%和49.4%;Transwell细胞迁移实验显示COL6A1-siRNA组细胞迁移能力明显减弱(P＜0.05),穿膜细胞数为空白对照组的45.3%。结论:利用siRNA技术能有效降低COL6A1基因的表达,同时有效抑制T24细胞的体外增殖和侵袭的能力。
- 膀胱癌 /
- COL6A1 /
- 增殖 /
- 侵袭 /
- RNA干扰
Abstract: Objective: To examine the influence of COL6 A1 on proliferation and invasion of bladder carcinoma cell. Method: The expression of COL6 A1 gene was knocked down using RNA interference (RNAi) in the bladder carcinoma cell T24. The transcription level of COL6 A1 was tested by RT-PCR. The expression of COL6 A1 protein was examined by Western blotting. The cell proliferation was detected by MTT. The cell's invasion ability was performed on Matrigel transwell assay. Result: The results showed that the level of COL6 A1 mRNA and protein expression were significantly reduced after RNAi with 20.5% of control group on mRNA expression and 14.1% of control group on protein expression. MTT assay showed that the proliferation of T24 cell was decreased due to RNA interference, with OD values reaching 75.6%, 58.2% and 49.4% of control group at 1 d, 2 d and 3 d respectively. Transwell assay showed that invasion abilities of T24 cells were reduced obviously due to COL6 A1 RNAi (P＜0.05), the transmembrane cells were 54.3% of control group.Conclusion: Our findings revealed that down-regulated COL6 A1 could inhibit abilities of proliferation and invasion in T24 cells. COL6 A1 can serve as a potential target for gene therapy of human bladder carcinoma.
- bladder carcinoma /
- COL6A1 /
- proliferation /
- invasion /
- RNA interference

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