RNA干扰Star3基因对EC-1细胞侵袭转移和增殖能力的影响
Influence of Stat3 gene repression by RNA interference on metastasis and proliferation of EC-1 cells
轩小燕 1李珊珊 2郑献召 2李娜 2王丰 2高远 2张红燕 2闫爱华2
作者信息
- 1. 河南省肿瘤病理重点实验室,郑州大学第一附属医院病理科;河南省肿瘤病理重点实验室,郑州大学第一附属医院病理科,郑州大学基础医学院微生物学与免疫学教研室,河南,郑州,450052
- 2. 河南省肿瘤病理重点实验室,郑州大学第一附属医院病理科
- 折叠
摘要
目的:构建针对信号转导子和转录激活子3(Stat3)基因的siRNA表达载体,研究Stat3基因沉默对人食管鳞癌EC-1细胞侵袭转移及增殖的影响.方法:构建pSUPER-EG-FP-Stat3 siRNA表达载体,转染EC-1细胞,RT-PCR,WesternBlot检测Stat3的表达;Boyden-chamber体外侵袭实验检测siRNA对细胞侵袭转移能力的影响,MTT法检测siRNA对细胞增殖能力的作用.结果:酶切和测序证实pSUPER-EGFP-Stat3 siRNA表达载体构建成功;RT-PCR,Western Blot结果表明,siRNA能有效地降解EC-1细胞中Stat3基因的mRNA,下调Stat3蛋白的表达;转染siRNA后,与对照组相比,EC-1细胞侵袭转移和增殖能力明显下降.结论:成功构建针对Stat3基因的siRNA表达载体,可有效沉默Stat3基因在食管鳞癌EC-1细胞中的表达,抑制EC-1细胞的侵袭转移及增殖能力,可为食管癌的基因治疗提供理论依据.
Abstract
AIM: To construct the siRNA expression vector targeting human Stat3 gene and to detect its silencing effects in esophageal carcinoma EC-1 cell lines. METHODS: siRNA expression vectors pSUPER-EGFP-Stat3 were constructed and were transfected into EC-1 cells. The expression of Stat3 mRNA and protein were detected by RT-PCR and Western blot. The invasion ability of EC-1 cells was determined by Boyden-chamber model in vitro and MTT assay was applied to determine the proliferation of EC-1 cells. RESULTS: The siRNA expression vectors pSUPER-EGFP-Stat3 were confirmed by enzyme digestion and DNA sequencing analysis. The results of RT-PCR and Western blot indicated that siRNA knocked down Stat3 mRNA and protein expression. Compared with the EC-1 cells in control groups,Boyden-chamber model showed that the invasion ability of EC-1 cells decreased significantly in interferential group (P < 0.05 ). The level of proliferation also declined in interferential group (P < 0.05). CONCLUSION: siRNA expression vectors inhibit the expression of Stat3 gene effectively in esophageal carcinoma EC-1 cells and suppress the invasion and proliferation ability of EC-1 cells,which lays the foundation for gene therapy of esophageal carcinoma.
关键词
Stat3基因/RNA干扰/EC-1细胞/肿瘤转移/癌,鳞状细胞/食管肿瘤Key words
Stat3/RNA interference technique/EC1/metastasis/carcinoma,squamous/esophageal neoplasms引用本文复制引用
基金项目
河南省科技攻关项目(072102310042)
河南省科技攻关项目(0624410048)
出版年
2009
NETL
NSTL
万方数据