摘要
目的探讨RNA干扰CST1基因表达对结直肠癌细胞增殖、侵袭及信号转导和转录激活因子3(STAT3)信号通路的影响。方法Western blotting法检测人结直肠癌SW480、HCT116、Caco2和HT29细胞及正常结肠NCM460细胞中CST1的蛋白表达。以Lipofectamine^TM 2000为载体,将设计合成的CST1的特异性siRNA及阴性对照siRNA转染HCT116细胞(si-CST1组和NC组),并设置空白对照组。转染48 h,MTT法检测细胞活力,Transwell小室检测穿膜细胞数,Western blotting法检测STAT3、磷酸化信号转导和转录激活因子3(p-STAT3)、增殖细胞核抗原(PCNA)、基质金属蛋白酶-2(MMP-2)和MMP-9的蛋白表达。结果人结直肠癌SW480、HCT116、Caco2和HT29细胞中CST1蛋白的表达均显著高于正常结肠NCM460细胞(P<0.05)。si-CST1组HCT116细胞中CST1蛋白的表达显著低于空白对照组(P<0.05)。与NC组比较,si-CST1组细胞活力和侵袭能力显著降低,p-STAT3、PCNA、MMP-2和MMP-9蛋白的表达显著降低(P<0.05)。结论RNA干扰CST1基因表达可抑制结直肠癌细胞增殖和侵袭,其机制可能与下调STAT3信号通路有关。
Objective An attempt was made in this paper to investigate the effect of RNA interference with CST1 gene expression on proliferation,invasion and signal transduction and activator of transcription 3(STAT3)signaling pathway in colorectal cancer cells.Methods The protein expression of CST1 in human colorectal cancer SW480,HCT116,Caco2 and HT29 cells,and normal colon NCM460 cells was detected by using western blotting.Lipofectamine^TM 2000 was used as a carrier,the specific siRNA of CST1(si-CST1 group)and negative control siRNA(NC group)were transfected into HCT116 cells,and a blank control group was established.After 48 h of transfection,the cell viability was detected by the MTT assay,the number of transmembrane cells was detected by transwell and the expression of STAT3,phosphorylation signal transduction and transcription activator 3(p-STAT3),proliferating cell nuclear antigen(PCNA),matrix metalloproteinase-2(MMP-2),and MMP-9 was detected by using western blotting.Results The protein expression of CST1 in human colorectal cancer cells SW480,HCT116,Caco2,and HT29 was significantly higher than that in normal colon cells NCM460(P<0.05).The expression of CST1 protein in HCT116 cells in the si-CST1 group was significantly lower than that in the blank control group(P<0.05).Compared with the NC group,the cell viability of the si-CST1 group,the cell invasion ability,and the protein expression of p-STAT3,PCNA,MMP-2,and MMP-9 were significantly decreased(P<0.05).Conclusion RNA interference with CST1 gene expression may inhibit proliferation and invasion of colorectal cancer cells,whose mechanism may be related to down-regulation of STAT3 signaling pathway.
作者
徐湖波
刘云华
江应平
雷珍
陈友平
XU Hubo;LIU Yunhua;JIANG Yingping;LEI Zhen;CHEN Youping(Department of Gastroenterology,the First People's Hospital of Tianmen City,Tianmen 431700,China)
出处
《国际消化病杂志》
CAS
2019年第4期281-285,共5页
International Journal of Digestive Diseases
基金
湖北省自然科学基金(2014CFC1104)
