抑制STAT3基因表达对人宫颈癌HeLa细胞定植和侵袭的影响  

The Influence of Silencing STAT3 Gene Expressions on the Location and Metastasis in Human Cervical Carcinoma HeLa Cells

作　　者：吴维光[1] 陈亚琼[1] 葛红雨[1] 韩建秋[1] 王勇梅[1] 

机构地区：[1]武警医学院附属医院妇产科,天津300162

出　　处：《肿瘤基础与临床》2010年第1期13-16,共4页journal of basic and clinical oncology

摘　　要：目的采用RNA干扰(RNAi)技术抑制人宫颈癌HeLa细胞信号转导及转录活化因子3(STAT3)基因的表达,观察其对HeLa细胞定植和侵袭能力的影响。方法针对STAT3基因设计并合成编码小干扰RNA(siRNA)的寡核苷酸,克隆入pSilencer2.1-U6-neo质粒中,从而构建针对STAT3基因的短发夹状siRNA真核表达质粒,用脂质体法将其转染入人宫颈癌HeLa细胞。利用RT-PCR技术和Western blot方法分别检测STAT3 mRNA和蛋白表达水平;通过软琼脂克隆形成实验和体外侵袭实验检测HeLa细胞的定植和侵袭能力。结果针对STAT3基因的短发夹状siRNA真核表达质粒成功转染人宫颈癌HeLa细胞;转染成功的HeLa细胞的STAT3基因mRNA及蛋白表达均下降;HeLa细胞在软琼脂中形成的克隆数和穿透Matrigel膜的细胞数均明显减少。结论针对STAT3基因的短发夹状siRNA真核表达质粒通过下调STAT3基因表达,抑制宫颈癌细胞的定植和侵袭能力。Objective To explore the influence of silencing signal transducers and activators of transcription-3（STAT3） gene expressions by RNA interference（RNAi） on the location and metastasis of human cervical carcinoma HeLa cells.Methods The small interference RNA（siRNA） templates were designed based on STAT3 gene sequence and was cloned into pSilencer2.1-U6-neo vector.This recombinant plasmid was transfected into HeLa cells by liposome method.The Western blot and the RT-PCR were used to detect STAT3 protein and mRNA,respectively.The colony formation assay and the transwell cabin assay were performed to measure the location and metastasis of HeLa cells.Results The short hairpin siRNA plasmid targeting STAT3 was successfully constructed and transfected into HeLa cells.The expressions of STAT3 gene and protein in these HeLa cells decreased.At the same time,the number of colony formation in soft agar and cells penetrating matrigel also decreased.Conclusion The short hairpin siRNA plasmid targeting STAT3 could inhibit the location and metastasis of cervical carcinoma HeLa cells through suppressing the expression of STAT3.

关 键 词：宫颈癌 信号转导及转录活化因子3 RNA干扰 

分 类 号：R737.33[医药卫生—肿瘤] R730.231[医药卫生—临床医学]