PTEN基因表达对大细胞肺癌H661细胞生物学行为影响  

EFFECTS OF PTEN GENE EXPRESSION ON BIOLOGICAL BEHAVIORS OF LARGE CELL LUNG CANCER CELL LINE H661

作　　者：林高阳 高大登 袁方 刘振波 LIN Gaoyang, GAO Dadeng, YUAN Fang, LIU Zhenbo(Department of Thoracic Surgery, The Affiliated Qingdao Hiser Medical Center of Qingdao University, Qingdao 266033, Chin)

机构地区：[1]青岛大学附属青岛市海慈医疗集团心胸外科,山东青岛266033

出　　处：《精准医学杂志》2017年第6期660-663,共4页Journal of Precision Medicine

摘　　要：目的探讨PTEN基因表达对大细胞肺癌H661细胞生物学行为的影响。方法合成PTEN基因的siRNA,构建PTEN基因过表达质粒。采用Lipofectamine 2000转染H661细胞,采用Western-blot方法检测PTEN蛋白在肺大细胞癌H661细胞中的表达;采用划痕实验检测PTEN基因对肺大细胞癌H661细胞迁移能力的影响;采用Trans-well小室实验检测PTEN基因对肺大细胞癌H661细胞侵袭能力的影响;采用平板克隆形成实验检测PTEN基因对H661细胞增殖的影响。结果Western-blot方法检测结果显示,转染pPTEN(过表达PTEN基因)质粒组中PTEN表达增加,PTEN RNAi组PTEN表达降低。划痕实验结果显示,PTEN RNAi组H661细胞迁移能力增强,细胞计数较阴性对照组增多(t=2.05,P<0.05);pPTEN质粒组细胞迁移能力较对照组和PTEN RNAi组明显降低,差异有显著性(t=3.30,P<0.05)。Trans-well小室实验结果显示,PTEN RNAi组细胞侵袭能力较对照组增强(t=2.03,P<0.05);pPTEN质粒组较对照组侵袭能力降低(t=2.95,P<0.05)。平板克隆形成实验结果显示,PTEN RNAi组细胞增殖能力较对照组增强(t=3.15,P<0.05);PTEN质粒组较对照组增殖能力降低(t=2.07,P<0.05)。结论 PTEN基因表达与肺癌的侵袭迁移能力密切相关,PTEN基因有望成为治疗肺癌侵袭迁移的潜在靶点。Objective To investigate the effects of phosphatase and tensin homolog （PTEN） gene expression on the bio- logical behaviors of human large cell lung cancer cell line H661. Methods The small interfering RNA （siRNA） of the PTEN gene was synthesized and the plasmid overexpressing the PTEN gene was constructed. H661 cells were divided into three groups： control group, RNAi group, and pPTEN group. The control group was transfected with the empty vector, the RNAi group was transfected with siRNA, and the pPTEN group was transfected with the plasmid overexpressing the PTEN gene; transfection was performed using lipofectamine 2000. For the three groups, Western-blot assay was used to measure the expression of PTEN protein in H661 cells; the migration, invasion, and proliferation ability of H661 cells were evaluated by wound healing assay, Trans-well assay, and colony formation assay, respectively. Results The Western-blot results showed that the expression level of PTEN protein was higher in the pPTEN group and lower in the RNAi group than in the control group. The wound healing assay showed that the RNAi group had a higher migration ability and a significantly higher cell count than the control group （t = 2.05, P〈0.05） ; the pPTEN group had a significantly lower migration ability than the control group and the RNAi group （t=3.30,P〈0.05）. The Trans-well assay showed that the invasion ability was significantly higher in the RNAi group and lower in the pPTEN group than in the control group （t=2.03 ,P%0.05 ;t =2.95 ,P〈0.05）. The colony formation assay showed that the proliferation ability was sig- nificantly higher in the RNAi group and lower in the pPTEN group than in the control group （t =3.15,P%0.05;t =2.07,P〈0.05）. Conclusion The invasion and migration ability of lung cancer are closely associated with the expression of PTEN gene, so the PTEN gene is promising to be a potential target for the treatment of lung cancer.

关 键 词：癌 非小细胞肺 PTEN磷酸水解酶 肿瘤侵润 肿瘤转移 细胞增殖 

分 类 号：R730.26[医药卫生—肿瘤]