外源性PTEN对胰腺癌细胞ASPC-1增殖、侵袭及转移的影响  

Heterogeneous PTEN inhibits ASPC-1 cell proliferation, invasion and metastasis

作　　者：李宏宇[1] 李建军[2] 刘旭[1] 吴春燕[1] 赵佳钧[1] 陈延志[2] 郭晓钟[2] 

机构地区：[1]沈阳军区总医院消化内科,沈阳110840 [2]中国医科大学附属第一医院放疗科

出　　处：《中华胰腺病杂志》2012年第1期13-15,共3页Chinese Journal of Pancreatology

基　　金：辽宁省博士启动基金（20081044）;辽宁省教育厅基金（L2010627）

摘　　要：目的研究第10染色体同源丢失性磷酸酶-张力蛋白基因（PTEN）对胰腺癌细胞系ASPC-1细胞周期、细胞增殖、血管内皮生长因子（VEGF）和表皮生长因子受体（EGFR）蛋白表达、裸鼠成瘤及转移能力的影响。方法将含PTEN基因的质粒pE—PTEN转染ASPC-1细胞，以空质粒pE转染为对照，分别获得ASPC-1-pE-PTEN（A—pE—P）细胞及ASPC-1-pE（A—pE）细胞。应用RT—PCR检测细胞PTENmRNA表达；免疫细胞化学法检测PTEN、VEGF、EGFR蛋白表达；克隆形成实验观察克隆形成数；Transwell小室观察细胞侵袭能力；裸鼠皮下注射癌细胞法观察成瘤情况。结果与ASPC一1细胞比较，A—pE—P细胞的PTENmRNA表达增加179．3％，PTEN蛋白表达明显增加；VEGF蛋白表达明显减少；EGFR蛋白表达无明显变化；G2／M期细胞数明显增加[（31．5±1．76）％比（26．81±1．03）％，P〈0．05]；克隆形成数降低[（24．0±3．9）比（33．3±3．4），F=4．283，P〈0．01]，克隆形成率降低28％；穿膜细胞数明显减少[每高倍视野（46．3±6．6）比（63．8±7．5）个，F=2．476，P〈0．05]；种植瘤体积明显缩小[（142．4±30．9）比（202．7±43．6）mm^2，t=4．834，P〈0．01]，抑瘤率达42．4％；远处转移灶明显减少[（2．0±0．7）比（5．0±1．3）个，t=0．451，P〈0．01]。结论PTEN基因转染后ASPC-1细胞VEGF表达减少，细胞生长被阻滞在G2／M期，增殖及转移被抑制。Objective To investigate the effects of heterogeneous phosphatase and tensinhomologue deleted on chromosome ten （PTEN） on cell cycles, proliferation, invasion, tumorigenicity, metastasis and the expressions of vascular endothelial growth factor （VEGF） and epidermal growth factor receptor （EGFR） proteins in human pancreas cancer cell line （ ASPC-1 ）. Methods ASPC-1 cells was transfected with plasmid pE-PTEN containing PTEN, and empty plasmid pE-PTEN transfection was used as control, then ASPC-I-pE- PTEN （A-pE-P） cell and ASPC-l-pE （A-pE） cell was obtained. The expression of PTEN mRNA was determined by RT-PCR. FFEN, VEGF and EGFR proteins were measured by cell immunohistochemical method. Clone formation assay was used to observe the numbers of clone. Transwell was used to test the invasion ability of cells. The growths of tumor were detected by nude mice subcutaneous injection of cancer cells in vivo. Results Compared with ASPC-1, the expressions of PTEN mRNA of A-pE-P increased by 179.3% , and the expressions of PTEN protein were also significantly increased. The expressions of VEGF protein were significantly decreased. The expressions of EGFR protein were not significantly changed. Number of G2/M phase cells was significantly increased from （ 26.81± 1.03 ） % to （ 31.5 ± 1.76） % （ P 〈 0.05 ）. The numbers of clone was decreased by 28% （P 〈0.05）. The number of penetrating cells was decreased [（46.3±6.6） vs （63.8 ±7.5） per high power fiehl, P 〈0.051. The tumor volumes were signifieanlly reduced [ （ 142.4 ± 30.9 ） vs （202.7± 43.6） mm^3 , P 〈 0.05 ]. The tumor inhibitory rate was 42.4%. The distant metastases were significantly reduced [ （ 2.0 ± 0.7 ） vs （ 5.0 ± 1.3 ） , P 〈0.01 ]. Conclusions Heterogeneous PTEN ean not only inhibit the prolilferation, invasion and metastasis of ASPC-I cells, arrest the cell growth at G2/M phase, but also decrease the expressions of VEGF.

关 键 词：胰腺肿瘤 基因 肿瘤抑制 转染 第10染色体同源丢失性磷酸酶.张力蛋白基因 

分 类 号：R735.9[医药卫生—肿瘤]