Wu Wei-Guang, Yu Yue-Cheng, Chen Ya-Qiong, Cao Xiu-Qin
Department of Obstetrics & Gynecology, Affiliated Hospital, Medical College of Chinese People's Armed Police Forces, Tianjin 300162, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2008 Oct;24(10):986-8.
To investigate the effects of eukaryotic vector expressing short hairpin RNA (shRNA) of signal transducers and activators of transcription 3 (STAT3) on the proliferation and apoptosis of SiHa cells.
shRNA templates were designed based on STAT3 gene sequence and cloned into pSilencer2.1-U6-neo vector. The resultant plasmid was transfected into SiHa cells with Lipofectamine 2000. The STAT3 protein and mRNA were detected by Western blot and RT-PCR, respectively. The cellular growth activity was assayed by MTT and the apoptosis was tested by flow cytometry.
The plasmid pSilencer2.1-U6-neo-STAT3 was successfully constructed and transfected into SiHa cells. The expression of STAT3 in SiHa cells decreased, the cellular growth activity decreased, and the cell apoptosis increased.
The siRNA expressing plasmid pSilencer2.1-U6-neo-STAT3 can inhibit cellular proliferation and induce the apoptosis of cervical cancer cells by suppressing the expression of STAT3.
研究表达信号转导与转录激活因子3（STAT3）短发夹RNA（shRNA）的真核载体对SiHa细胞增殖和凋亡的影响。
根据STAT3基因序列设计shRNA模板，并克隆到pSilencer2.1-U6-neo载体中。将所得质粒用Lipofectamine 2000转染至SiHa细胞。分别通过蛋白质免疫印迹法和逆转录-聚合酶链反应检测STAT3蛋白和mRNA。用MTT法检测细胞生长活性，用流式细胞术检测细胞凋亡。
成功构建质粒pSilencer2.1-U6-neo-STAT3并转染至SiHa细胞。SiHa细胞中STAT3的表达降低，细胞生长活性降低，细胞凋亡增加。
表达小干扰RNA的质粒pSilencer2.1-U6-neo-STAT3可通过抑制STAT3的表达抑制细胞增殖并诱导宫颈癌细胞凋亡。
