靶向抑制磷脂酰肌醇3-激酶110β因子的表达对人胃癌细胞生长影响的实验研究  

Study on the effects of target inhibiting phosphoinositide 3-kinases p110beta on the growth of human gastric cancer cells

作　　者：许慈[1] 蔡懿婷[1] 董晴晴[1] 王碧君[1] 戴强[1] 朱黎明[1] 

机构地区：[1]上海交通大学医学院附属第三人民医院消化科,上海201999

出　　处：《中国医药导报》2016年第35期28-31,49,F0003,共6页China Medical Herald

基　　金：上海交通大学医学院新华集团基金(13XJ22012)

摘　　要：目的探讨沉默磷脂酰肌醇3-激酶(PI3K)p110β因子的表达对人胃癌细胞生长的影响。方法运用RNA干扰技术,合成靶向抑制PI3Kp110β的小干扰RNA(si RNA)序列并分别瞬时转染人胃癌细胞株AGS、SGC7901和MKN45(si-PI3Kp110β),作为实验组,未转染的细胞为空白对照组(Con),转染无义序列的si RNA为阴性对照组(si-NC)。采用RT-PCR和Western Blot方法验证瞬时转染后PI3Kp110β mRNA和蛋白在空白对照组(Con)、阴性对照组(si-NC)和实验组(si-PI3Kp110β)细胞中的表达水平,并通过MTT法检测细胞增殖及Annexin V-FITC/PI双染法检测AGS细胞凋亡率的情况。结果小分子干扰RNA(si RNA)瞬时转染人胃癌细胞株AGS、SGC7901和MKN45后,能有效下调PI3Kp110β在细胞内的表达水平。与阴性对照组(si-NC)比较,静默PI3Kp110β因子可减少AGS细胞的增殖(P<0.05),并明显增加AGS细胞的凋亡率(P<0.01)。结论小分子干扰RNA可有效下调PI3Kp在人胃癌细胞中的表达,沉默PI3Kp110β因子可有效抑制胃癌肿瘤细胞的增殖,促进细胞凋亡,提示针对PI3Kp110β蛋白的干预治疗有可能成为抑制胃癌细胞生长的一个新的治疗方法。Objective To study the effect of PI3Kp110β silencing on growth of human gastric cancer cells in vitro. Methods The small interfering RNA sequence targeted by PI3Kp110β and negative sequence was transfected into gas- tric carcinoma cells including AGS, SGC7901 and MKN45 for 48 h respectively by RNA interfering （si-PI3Kp110β group）. Other two groups include untransfected cells （Con group） and nonsense siRNA sequence transfected cells （si- NC group）. The mRNA and protein expression of PI3Kp110β were determined by reverse transeriptase polymerase chain reaction （RT-PCR） and Western blotting. The cellular proliferation was measured by MTT assay, and cell apop- tosis was determined by flow cytometry in AGS cells. Results The expression level of PI3Kp110β was downregulated significantly in si-PI3Kp110β group in three human gastric carcinoma cell lines AGS, SGC7901 and MKN45. The cell proliferation was decreased （P 〈 0.05） and cell apoptosis was significantly increased （P 〈 0.01） in si-PI3Kp110β group compared with si-NC group in AGS cells. Conclusion The expression of PI3Kp110β in gastric cancer cells is effec- tively decreased with RNA interference. Silencing PI3Kp110β expression can effectively inhibit the proliferation of gas- tric carcinoma cells and induce cell apoptosis, siRNA targeting PI3Kp110β may become one of the methods to treat gastric carcinoma.

关 键 词：磷脂酰肌醇3-激酶p110β RNA干扰 胃肿瘤 增殖 凋亡 

分 类 号：R734[医药卫生—肿瘤]