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.docVIP

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 　　[摘要] 目的 探?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（siRNA）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siRNA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（siRNA）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 　　[关键词] 磷脂酰肌醇3-激酶p110β； RNA干扰；胃肿瘤；增殖；凋亡 　　[中图分类号] R734 [文献标识码] A [文章编号] 1673-7210（2016）12（b）-0028-05 　　Study on the effects of target inhibiting phosphoinositide 3-kinases p110beta on the growth of human gastric cancer cells 　　XU Ci CAI Yiting DONG Qingqing WANG Bijun DAI Qiang ZHU Liming▲ 　　Department of Gastroenterology， No.3 People′s Hospital Affiliated to Shanghai Jiaotong University School of Medicine， Shanghai 201999， China 　　[Abstract] 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 gastric 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 transcriptase polymerase chain reaction （RT-PCR） and Western blotting. The cellular proliferation was measured by MTT assay， and cell apoptosis 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 　　胃癌是常见的消化道肿瘤，在我国胃癌患者5年生存率低，寻找关键的肿瘤信号调节因子，可为其治疗带来突破。研究发现，磷脂酰肌醇-3激酶（PI3K）/蛋白激酶B（Akt）信号通路在肿瘤转移以及耐药中发挥重要作用[1]。PI3K家族分为3种亚型，深入研究的是ⅠA型PI3K，由p