Suppression of TSPAN1 by RNA interference inhibits proliferation and invasion of colon cancer cells in vitro
Suppression of TSPAN1 by RNA interference inhibits proliferation and invasion of colon cancer cells in vitro
复制标题
RNA干扰抑制TSPAN1抑制结肠癌细胞的体外增殖和侵袭
DOI：
10.1177/030089161009600517
复制
发表时间：
2010-09-01
期刊：
影响因子：
1.9
通讯作者：
Zhu, Jianwei
中科院分区：
文献类型：
--
作者：
Chen, Li;Yuan, Daiyue;Zhu, Jianwei
目标和背景。研究RNA干扰（RNAi）下调TSPAN 1表达对人结肠癌细胞体外增殖和侵袭的影响。使用基于载体（pU 6 H1-GFP）的小干扰RNA（siRNA）质粒基因沉默系统进行RNAi，以特异性敲低结肠癌细胞系HCT-8中的TSPAN 1表达。逆转录聚合酶链反应检测TSPAN 1 mRNA的表达。使用Western印迹和免疫荧光显微镜观察TSPAN 1蛋白表达。MTT法检测细胞增殖活性，流式细胞仪检测细胞周期。HCT-8细胞的侵袭能力使用双培养室进行检测，所述双培养室由涂覆有Matrigel（8.0 μ m孔径）的聚碳酸酯膜分隔。转染TSPAN 1 siRNA质粒后，TSPAN 1 mRNA和蛋白表达显著降低。mRNA和蛋白质的减少与TSPAN 1荧光染色的显著减少和由于细胞周期停滞在G1/G 0期而导致的细胞增殖的减少相关。这些变化与HCT-8细胞的侵袭数量显著减少有关。RNAi介导的TSPAN 1表达下调在体外显著抑制结肠癌细胞的增殖和侵袭。这一发现表明TSPAN 1在结肠癌进展中起重要作用，靶向TSPAN 1的RNAi可能是治疗结肠癌的潜在治疗策略。可在www.tumorionline.it上获得免费全文
Aims and background. To investigate effect of TSPAN1 downregulation by RNA interference (RNAi) on proliferation and invasion of human colon cancer cells in vitro.Methods and study design. RNAi was performed using the vector (pU6H1-GFP)-based small-interfering RNA (siRNA) plasmid gene silencing system to specifically knock down TSPAN1 expression in a colon cancer cell line, HCT-8. The expression of TSPAN1 mRNA was detected by reverse-transcription polymerase chain reaction. TSPAN1 protein expression was observed using Western blots and immunofluorescent microscopy. Cell proliferation and cell cycle assay were measured using methyl thiazolyl tetrazolium (MTT) and flow cytometry, respectively. The invasive ability of HCT-8 cells was examined using a duel culture chamber separated by polycarbonate membranes coated with Matrigel (8.0-mu m pore size).Results. After transfection with the TSPAN1 siRNA plasmid, TSPAN1 mRNA and protein expression was significantly decreased. The decrease in mRNA and protein was associated with a significant decrease in TSPAN1 fluorescent staining and a decrease in cell proliferation due to cell cycle arrest in the G1/G0 phase. A significant decrease in the number of invading HCT-8 cells was associated with these changes.Conclusion. RNAi-mediated downregulation of TSPAN1 expression significantly inhibits the proliferation and invasion of colon cancer cells in vitro. This finding suggests that TSPAN1 plays an important role in colon cancer progression, and RNAi targeting of TSPAN1 may be a potential therapeutic strategy for the treatment of colon cancer. Free full text available at www.tumorionline.it