摘要
目的选择PTEN低表达胰腺癌细胞系,探讨体外转染PTEN对该细胞系的影响。方法将PTEN转染到PTEN低表达胰腺癌细胞系,应用RTPCR、免疫组化、克隆形成率及观察裸鼠移植瘤生长等方法检测PTEN对胰腺癌细胞系的影响。结果PTEN在ASPC1细胞呈低表达,ASPC1、ApE、ApEP细胞PTENmRNA表达量和克隆形成率分别为203%、150%、568%和333%、317%、240%;ASPC1细胞转染后PTEN蛋白表达水平升高,血管内皮生长因子蛋白下降,表皮生长因子受体蛋白无明显变化;5周时转染前后荷瘤裸鼠瘤结节体积分别为2027mm3和1424mm3,差异有统计学意义(P<001)。结论PTEN低表达ASPC1细胞系经转染PTEN后细胞增殖活性及裸鼠致瘤能力明显降低。
Objective To investigate the effect of phosphatase and ensinhomology deleted on chromosome ten( PTEN) on the human pancreatic cancer cell line without or low PTEN expression.Methods We selected the lowest PTEN gene expressive pancreatic cancer cell line among the four pancreatic cancer cell lines (Miapaca I, Miapaca II, JF305 and ASPC-1) through RT-PCR assay method, and transfected plasmid (Peak8) inserting PTEN or not in vitro into it by lipofectin. The effect of PTEN transfection on the lowest PTEN gene expressive pancreatic cancer cell line was carried out by flow cytometry, immunohistochemical staining, cloning survival assay, as well as tumorigenicity in nude mice. Results The lowest PTEN gene expression was found in ASPC-1 cells. PTEN mRNA expression and cloning plating efficiency in the ASPC-1, A-pE and A-pE-P cells were 20.3%, 15.0 %, 56.8 % and ~33.3 %, 31. 7 %, 24.0 % respectively. We found that ASPC-1 cells transfected with PTEN exhibited significantly more protein, less vascular endothelial growth factor protein, and non-changed epidermal growth factor receptor protein comparing with vector-transfected cells. The tumor volumes were 202.7 mm3 and 142.4 mm3, pre-and post-transfection with significance(P<0.01) within 5 weeks.Conclusions Our findings suggested that PTEN mRNA exhibited the lowest expression in ASPC-1 cell line, exogenous PTEN dramatically inhibits the growth of ASPC-1 cells transfected with PTEN gene in nude mice.
出处
《中华内科杂志》
CAS
CSCD
北大核心
2005年第3期191-194,共4页
Chinese Journal of Internal Medicine
