Xie Xiao-bin, Long Jie, Yang Fang, Zhang Ya-jie
Department of Pathology, Guangzhou Medical College, Guangzhou 510182, China.
Zhonghua Bing Li Xue Za Zhi. 2009 Jul;38(7):472-6.
To study the effect of short haipin RNA (shRNA) on expression of vascular endothelial growth factor C (VEGF-C) and proliferation and invasion behavior of human breast cancer cell MCF-7.
The recombinant vector (pSIREN-VEGF-C) was transfected into the human breast cancer cell MCF-7 by liposome and the positive transfected cell clones were screened with puromycin. Expression of VEGF-C in MCF-7 cells after gene transfer was detected by real-time quantitative PCR and Western blot assay, respectively. Proliferation and invasion ability of transfected cells were analyzed by MTT and Transwell filter.
The expressions of VEGF-C mRNA and protein were decreased markedly compared with the control group after the transfection and the inhibitive ratio was 95% and 100% respectively (P<0.05). The proliferation of MCF-7 cells transfected by pSIREN-VEGF-C, measured with MTT assays, was significantly decended (P<0.05). The invasion ability of passing through the Transwell filter of MCF-7 cells transfected by pSIREN-VEGF-C were declined evidently (P<0.05).
The recombinant vector (pSIREN-VEGF-C) have been proved not only to be effective and specific for down-regulation of VEGF-C, but also can inhibit the proliferation and invasion of MCF-7 cells significantly.
研究短发夹RNA（shRNA）对人乳腺癌细胞MCF-7血管内皮生长因子C（VEGF-C）表达及增殖和侵袭行为的影响。
采用脂质体将重组载体（pSIREN-VEGF-C）转染至人乳腺癌细胞MCF-7，并用嘌呤霉素筛选阳性转染细胞克隆。分别采用实时定量PCR和蛋白质免疫印迹法检测基因转染后MCF-7细胞中VEGF-C的表达。通过MTT法和Transwell小室分析转染细胞的增殖和侵袭能力。
转染后VEGF-C mRNA和蛋白表达与对照组相比明显降低，抑制率分别为95%和100%（P<0.05）。用MTT法检测，pSIREN-VEGF-C转染的MCF-7细胞增殖明显下降（P<0.05）。pSIREN-VEGF-C转染的MCF-7细胞穿过Transwell小室的侵袭能力明显下降（P<0.05）。
已证实重组载体（pSIREN-VEGF-C）不仅能有效、特异地下调VEGF-C，还能显著抑制MCF-7细胞的增殖和侵袭。
