Li Hong-yu, Yu Jie-ping, Guo Xiao-zhong, Li Jian-jun, Zhao Jia-jun, Wang Di, Huo Ran, Xia Yu-ting
Department of Gastroenterology, Shenyang General Hospital of PLA, Shenyang 110016, China.
Zhonghua Nei Ke Za Zhi. 2005 Mar;44(3):191-4.
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.
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.
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 mm(3) and 142.4 mm(3), pre-and post-transfection with significance (P < 0.01) within 5 weeks.
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.
研究第10号染色体缺失的磷酸酶及张力蛋白同源物（PTEN）对PTEN无表达或低表达的人胰腺癌细胞系的影响。
通过逆转录聚合酶链反应（RT-PCR）检测方法，在4种胰腺癌细胞系（Miapaca I、Miapaca II、JF305和ASPC-1）中筛选出PTEN基因表达最低的胰腺癌细胞系，并在体外通过脂质体将插入或未插入PTEN的质粒（Peak8）转染至该细胞系。通过流式细胞术、免疫组织化学染色、克隆存活试验以及裸鼠成瘤性试验，研究PTEN转染对PTEN基因表达最低的胰腺癌细胞系的影响。
ASPC-1细胞中PTEN基因表达最低。ASPC-1、A-pE和A-pE-P细胞中PTEN mRNA表达及克隆铺板效率分别为20.3%、15.0%、56.8%和33.3%、31.7%、24.0%。我们发现，与载体转染细胞相比，转染PTEN的ASPC-1细胞表现出更多的蛋白、更少的血管内皮生长因子蛋白，而表皮生长因子受体蛋白无变化。转染前后5周内肿瘤体积分别为202.7 mm³和142.4 mm³，差异有统计学意义（P < 0.01）。
我们的研究结果表明，PTEN mRNA在ASPC-1细胞系中表达最低，外源性PTEN显著抑制裸鼠中PTEN基因转染的ASPC-1细胞的生长。
