Gao Zhen, Huang Chen, Qiu Zheng-Jun, Jiang Tao, Zhu Lin, Cao Jun, Zhang Fang, Huang Ke-Jian
Department of General Surgery, Affiliated First People's Hospital, Shanghai Jiaotong University, Shanghai 200080, China.
Zhonghua Wai Ke Za Zhi. 2008 Jul 1;46(13):1010-3.
To investigate the effect of RNAi-mediated STAT3 gene inhibition on metastasis of human pancreatic cancer cells and its underlying mechanism.
STAT3 shRNA expression vector was constructed and transfected into SW1990 cells. STAT3 mRNA and STAT3 DNA-binding activity were examined using reverse transcription polymerase chain reaction (RT-PCR) and electrophoretic mobility shift assay (EMSA), respectively. The metastasis ability of SW1990 cells in vivo was determined in acute hematogenous metastasis model using nude mice. RT-PCR was performed to detect the mRNA expression of the MMP-2 and VEGF.
The mRNA expression of STAT3 and STAT3 DNA-binding activity were inhibited significantly by stable transfection of STAT3 shRNA expressing vectors. RNAi inhibition of STAT3 significantly suppressed the metastasis ability of SW1990 cells in vivo, and also markedly reduced the mRNA expression of MMP-2 and VEGF in SW1990 cells.
Inhibition of STAT3 by RNAi significantly inhibits the metastasis ability of pancreatic cancer cells through down-regulation of MMP-2 and VEGF, and may provide a novel strategy for preventing the metastasis of pancreatic cancer.
探讨RNA干扰介导的信号转导子和转录激活子3（STAT3）基因抑制对人胰腺癌细胞转移的影响及其潜在机制。
构建STAT3短发夹RNA（shRNA）表达载体并转染至SW1990细胞。分别采用逆转录聚合酶链反应（RT-PCR）和电泳迁移率变动分析（EMSA）检测STAT3信使核糖核酸（mRNA）和STAT3 DNA结合活性。利用裸鼠急性血行转移模型测定SW1990细胞在体内的转移能力。进行RT-PCR以检测基质金属蛋白酶-2（MMP-2）和血管内皮生长因子（VEGF）的mRNA表达。
稳定转染STAT3 shRNA表达载体可显著抑制STAT3的mRNA表达和STAT3 DNA结合活性。RNA干扰抑制STAT3可显著抑制SW1990细胞在体内的转移能力，同时也显著降低SW1990细胞中MMP-2和VEGF的mRNA表达。
RNA干扰抑制STAT3可通过下调MMP-2和VEGF显著抑制胰腺癌细胞的转移能力，可能为预防胰腺癌转移提供新策略。
