生长抑素2型受体基因转染对胰腺癌细胞蛋白表达的影响

目的 探讨生长抑素2型受体(hSSTR2)基因转染对胰腺癌细胞PANC1蛋白表达的影响,以寻找新的胰腺癌敏感治疗靶点.方法 利用前期构建的腺病毒载体Ad.CMV.hSSTR2.GFP将hSSTR2全长cDNA导入胰腺癌细胞PANC1.采用双向荧光差异凝胶电泳(2D-DIGE)技术分离并筛选转染hSSTR2的实验组、空载体对照组以及空白组胰腺癌细胞之间差异表达蛋白,用反射式基质辅助激光解吸附电离串联飞行时间质谱(MALDI-TOF/TOF)技术对差异蛋白进行鉴定.结果 hSSTB2成功地转染了胰腺癌细胞,获得了hSSTR2阴性和阳性表达的PANC1荧光差异蛋白表达图谱,经DeCyder v6.5软件分析,共有18个差异在1.3倍以上的蛋白质点,经质谱鉴定得到13个蛋白质.低表达的蛋白7个,为GMP合酶、磷酸化应激诱导蛋白、谷氨酸脱氢酶、Septin-11、波形蛋白、异柠檬酸脱氢酶α亚基、线粒体内膜易位酶;高表达蛋白6个,为真核延长因子1α1、丙酮酸激酶异构体M2型、烯酰-CoA水合酶、转录调节因子1-β、Mitofilin、HSP105.结论 hSSTR2基因转染胰腺癌细胞PANC1后引起蛋白表达发生变化,这些差异蛋白功能涉及到糖、脂肪、核酸代谢以及细胞生长调节和细胞凋亡等,从而为寻找新的胰腺癌敏感治疗靶点奠定基础.

Effects of hSSTR2 gene in vitro transfection on differential proteins expression in pancreatic cancer cell line Panc-1

Objective To study the effects of hSSTR2 gene in vitro transfection on differential proteins expression in pancreatic cancer cell line Panc-1 and search new sensitive therapeutic targets of pancreatic cancer. Methods The full length hSSTR2 cDNA was introduced into pancreatic cancer cell line Panc-1 by adenovirus vector ( Ad. CMV. hSSTR2. GFP) mediated transfection. The differential expressed proteins between the hSSTR2 transfection group, vector control and mock control were isolated and screened by 2D-DIGE analysis. Protein identification was performed by peptide mass finger printing with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/TOF). Results The hSSTR2 gene was transfected into Panc-1 pancreatic cancer cells in vitro successfully, and fluorescence difference protein expression patterns were established between hSSTR2 negative and positive expression of Panc-1 cell. Analysis by DeCyder v6.5 software showed a total of 18 protein spots ( > 1.3-fold) and these protein spots were identified by mass spectrometry as 13 proteins. Proteins with lower abundance levels included GMP synthase, stress induced phosphoprotein 1, glutamate dehydrogenase 1, Septin-11, vimentin, Isocitrate dehydrogenase [NAD] subunit alpha, Import inner membrane translocase subunit TIM50. Proteins with high abundance levels included Elongation factor 1-alpha-1, Isoform M2 of Pyruvate kinase isozymes M1/M2, Enoyl-CoA hydratase,tripartite motif-containing 28 protein, Mitofilin, HSP105. Conclusions The proteins expression changed after hSSTR2 gene in vitro transfection in Panc-1 cells, and the function of difference proteins involved the process of metabolism of sugar, fat and nucleic acid, and the regulation of cell growth. The present study paved the way for searching new sensitive therapeutic targets of pancreatic cancer.