CAJ | 학술논문

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PTEN mRNA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-1细胞比较,A-pE-P细胞的PTEN mRNA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)mm3,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.
목적 연구제10염색체동원주실성린산매-장력단백기인(PTEN)대이선암세포계ASPC-1세포주기、세포증식、혈관내피생장인자(VEGF)화표피생장인자수체(EGFR)단백표체、라서성류급전이능력적영향.방법 장함PTEN기인적질립pE-PTEN전염ASPC-1세포,이공질립pE전염위대조,분별획득ASPC-1-pE-PTEN (A-pE-P)세포급ASPC-1-pE (A-pE)세포.응용RT-PCR검측세포PTEN mRNA표체；면역세포화학법검측PTEN、VEGF、EGFR단백표체；극륭형성실험관찰극륭형성수；Transwell소실관찰세포침습능력；라서피하주사암세포법관찰성류정황.결과 여ASPC-1세포비교,A-pE-P세포적PTEN mRNA표체증가179.3％,PTEN단백표체명현증가；VEGF단백표체명현감소；EGFR단백표체무명현변화；G2/M기세포수명현증가[(31.5±1.76)％비(26.81±1.03)％,P＜0.05]；극륭형성수강저[(24.0±3.9)비(33.3 ±3.4),F=4.283,P＜0.01],극륭형성솔강저28％；천막세포수명현감소[매고배시야(46.3±6.6)비(63.8±7.5)개,F=2.476,P＜0.05]；충식류체적명현축소[(142.4±30.9)비(202.7±43.6)mm3,t=4.834,P＜0.01],억류솔체42.4％；원처전이조명현감소[(2.0±0.7)비(5.0±1.3)개,t =0.451,P＜0.01].결론 PTEN기인전염후ASPC-1세포VEGF표체감소,세포생장피조체재G2/M기,증식급전이피억제.
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-1-pE-PTEN (A-pE-P) cell and ASPC-1-pE (A-pE) cell was obtained.The expression of PTEN mRNA was determined by RT-PCR. PTEN,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 field,P ＜0.05].The tumor volumes were significantly reduced [(142.4 ±30.9) vs (202.7 ±43.6) mm3,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 can not only inhibit the proliferation,invasion and metantasis of ASPC-1 cells,arrest the cell growth at G2/M phase,but also decrease the expressions of VEGF.