文章摘要

Fascin shRNA 通过Notch1 信号通路调控结直肠癌细胞的生长

作者: 1余 伟, 1张 志标, 1马 达
1 汉川市人民医院普外二科,湖北 汉川 431600
通讯: 张 志标 Email: 1557666150@qq.com
DOI: 10.3978/j.issn.2095-6959.2019.02.002

摘要

目的:研究聚束蛋白(Fascin)shRNA对结直肠癌细胞生长的影响。方法:结直肠癌细胞HT29感染Fascin shRNA和shRNA control慢病毒,real-time PCR和Western印迹法分别测定细胞中Fascin mRNA和蛋白的表达水平,同时用Western印迹法检测细胞中的Notch1蛋白水平。用Notch1信号通路抑制剂DAPT处理下调Fascin后的HT29细胞,MTT测定细胞增殖,流式细胞术检测细胞周期,Western印迹法检测周期蛋白p27、细胞周期蛋白D1(Cyclin D1)表达水平。结果:Fascin shRNA慢病毒感染后的细胞中Fascin转录和表达水平均降低,而shRNA control慢病毒感染对细胞中的Fascin表达水平没有影响。沉默Fascin的细胞增殖能力降低,细胞G0/G1比例升高,细胞中Cyclin D1蛋白水平降低,p27蛋白水平升高。沉默Fascin能够降低细胞中Notch1蛋白水平,并且Notch1信号通路抑制剂DAPT可以协同沉默Fascin,进一步抑制结直肠癌细胞增殖,阻滞结直肠癌细胞周期,减少Cyclin D1蛋白表达,促进p27蛋白表达。结论:Fascin shRNA通过抑制Notch1信号通路阻碍结直肠癌细胞增殖,将细胞周期阻滞在G0/G1期。
关键词: 结直肠癌;增殖;Notch1信号通路;Fascin

Fascin shRNA regulates the growth of colorectal cancer cells through Notch1 signaling pathway

Authors: 1YU Wei, 1ZHANG Zhibiao, 1MA Da
1 Second Department of General Surgery, Hanchuan People’s Hospital, Hanchuan Hubei 431600, China

CorrespondingAuthor:ZHANG Zhibiao Email: 1557666150@qq.com

Abstract

Objective: To investigate the effect of Fascin shRNA on the grow th of colorectal cancer cells. Methods: Colorectal cancer cells HT29 infected with Fascin shRNA and shRNA control lentivirus, real-time PCR and Western blot were used to measure the expression of Fascin mRNA and protein, at the same time, the level of Notch1 protein was detected by Western blot, Notch1 signaling pathway inhibitor DAPT was used to the HT29 cells after downregulate Fascin. Cell proliferation, cell cycle, the expression levels of cyclin p27 and Cyclin D1 were detected by MTT assay, flow cytometry, and Western blot, respectively. Results: After Fascin shRNA lentivirus infection, Fascin transcription and expression levels were reduced. However, shRNA control lentivirus had no effect on Fascin expression. The cell proliferation ability of silent Fascin was reduced, the proportion of G0/G1 in cells increased, the level of Cyclin D1 protein in the cells decreased, the level of p27 protein was increased. Silencing Fascin reduced the level of Notch1 protein in cells. Moreover, Notch1 signaling pathway inhibitor DAPT coexisted with silencing Fascin to further inhibit the proliferation of colorectal cancer cells, the cell cycle of colorectal cancer was blocked, the expression of Cyclin D1 protein reduced, the expression of p27 protein promoted. Conclusion: Fascin shRNA can inhibit proliferation and blocked cell cycle of colorectal cancer cells by inhibiting Notch1 signaling pathway.
Keywords: colorectal cancer; proliferation; Notch1 signaling pathway; Fascin