慢病毒介导的生长抑素shRNA对BHK-21细胞生长的影响

The effects of lentviral-directed shRNA targeting somatostatin on cell proliferation of BHK-21 cells

采用慢病毒介导的shRNA沉默细胞自身生长抑素(SS)的表达,同时,以pcDNA3.1-SS(pSS)真核表达载体转染细胞作为阳性对照,研究SS对BHK-21细胞的抑制增殖作用,同时观察SS是否具有促进细胞凋亡的作用。MTT法绘制细胞生长曲线可知,pSS转染细胞的生长受到明显抑制,抑制效率为9.63%(P<0.05);而LV-sh2组细胞的生长密度是对照组细胞的117.33%(P<0.05),表明SS对细胞的增殖具有抑制作用。流式细胞检测细胞凋亡表明,pSS转染组和LV-shRNA感染组凋亡细胞含量分别是对照组凋亡细胞含量的1.97倍(P<0.05)和24.30%(P<0.05),表明SS通过诱导细胞凋亡发挥抑制细胞增殖的作用。本研究为SS及其类似物作为治疗药物的进一步开发应用提供了理论依据。

Several lines of evidence implicated that somatostatin（SS） is commonly expressed in many cells and has the anti-proliferative effects of cell growth,but the functions of the endogenous SS on the proliferation and apoptosis of normal cells are still largely Lrnknown. In this research, the endogenous SS expression was down-regulated in BHK-21 cells using a Lentiviral-shRNA（Lv） to examine whether it would have an effect on cell proliferation with the pcD- NA3.1-SS（pSS） transfected cells as the positive Control. Cell growth curve measured by using the MTT assay showed that higher levels of SS inhibit the cell growth ability significantly（9.63%, P〈0.05）. Reversely, lowering SS could stimulate the cell growth ability（17.33% ,P〈0.05）. In addition,the apoptosis of cells detected by flow cytometry indicated that lower percents （24. 30 %, P〈 0. 05） of apoptosis cells were observed relative to controlled BHK-21 cells, and higher percents were showed in pSS transfected cells（1.97 folds,P〈0.05）, which indicated that the anti-proliferative function of SS is via the induction of the cell apoptosis. This research gets theoretical foundation for SS and its analogs used as a potential drug of inhibitor in humans or animals.