CAJ | 학술논문

目的探讨第10号染色体上磷酸酶和张力蛋白同源缺失的基因(PTEN)对人卵巢癌A2780细胞侵袭和迁移能力的影响及相关机制.方法将携带外源性人PTEN基因的野生型质粒(WT-PTEN)和磷酸酶域突变型质粒(C124A-PTEN)转入人卵巢癌细胞株A2780中,利用Transwell小室法和划痕愈合实验检测转染前后细胞侵袭和迁移能力的变化,应用Western blot检测各组细胞PTEN及其下游相关蛋白表达的变化.以空载体质粒和未转染细胞作为对照.结果细胞侵袭实验显示,WT-PTEN/A2780、C124A-PTEN/A2780、GFP/A2780和A2780细胞穿透Matrigel胶的细胞数,分别为(24.3±2.5)个、(43.7±3.8)个、(44.7±2.1)个和(45.0±3.0)个,WT-PTEN/A2780细胞的穿膜数明显少于其他各组(P＜0.05).划痕愈合实验显示,WT-PTEN/A2780、C124A-PTEN/A2780、GFP/A2780和A2780细胞的迁移距离分别为(54.1±3.7)μm、(78.7±3.4)μm、(78.1±3.1)μm和(76.8±3.5)μm,WT-PTEN/A2780细胞的迁移速度明显慢于C124A-PTEN/A2780、GFP/A2780和A2780细胞(P＜0.05).WT-PTEN/A2780细胞中基质金属蛋白酶9(MMP-9)蛋白的表达明显低于C124A-PTEN/A2780、GFP/A2780和A2780细胞(P＜0.05).结论野生型PTEN可有效抑制A2780细胞的侵袭和迁移,该抑制作用与A2780细胞中MMP-9表达下调有关.
목적 탐토제10호염색체상린산매화장력단백동원결실적기인(PTEN)대인란소암A2780세포침습화천이능력적영향급상관궤제.방법 장휴대외원성인PTEN기인적야생형질립(WT-PTEN)화린산매역돌변형질립(C124A-PTEN)전입인란소암세포주A2780중,이용Transwell소실법화화흔유합실험검측전염전후세포침습화천이능력적변화,응용Western blot검측각조세포PTEN급기하유상관단백표체적변화.이공재체질립화미전염세포작위대조.결과 세포침습실험현시,WT-PTEN/A2780、C124A-PTEN/A2780、GFP/A2780화A2780세포천투Matrigel효적세포수,분별위(24.3±2.5)개、(43.7±3.8)개、(44.7±2.1)개화(45.0±3.0)개,WT-PTEN/A2780세포적천막수명현소우기타각조(P＜0.05).화흔유합실험현시,WT-PTEN/A2780、C124A-PTEN/A2780、GFP/A2780화A2780세포적천이거리분별위(54.1±3.7)μm、(78.7±3.4)μm、(78.1±3.1)μm화(76.8±3.5)μm,WT-PTEN/A2780세포적천이속도명현만우C124A-PTEN/A2780、GFP/A2780화A2780세포(P＜0.05).WT-PTEN/A2780세포중기질금속단백매9(MMP-9)단백적표체명현저우C124A-PTEN/A2780、GFP/A2780화A2780세포(P＜0.05).결론 야생형PTEN가유효억제A2780세포적침습화천이,해억제작용여A2780세포중MMP-9표체하조유관.
Objectiye To evaluate the effects of PTEN on invasive and migration ability of human ovarian cancer cell line A2780 and related mechanisms. Methods The plasmid including WT-PTEN and mutant PTEN were transferred into A2780 cells. The invasive and migration ability were measured before and after transfection by transwell chamber and wound-healing assays. The expression of PTEN protein and related proteins in the cells were detected by Western blot analysis. Empty plasmid-transfected A2780 and normal A2780 cells were used as control (the different four groups were named as WT-PTEN/A2780,C124A-PTEN/A2780, GFP/A2780 and A2780 ). Results The number of penetrating cells was significantly less in WT-PTEN/A2780 cells(24.3 ± 2.5 ) than those in C124A-PTEN/A2780, GFP/A2780 and A2780 cells (43.7 ± 3.8, 44.7 ± 2. 1 and 45.0 ± 3.0 ) ( P ＜ 0.05 ). The migration distance was markedly shorter in WT-PTEN/A2780 cells (54.1 ± 3.7) than those in C124A-PTEN/A2780, GFP/A2780 and A2780 cells (78.7 ± 3.4, 78.1 ± 3.1 and 76.8 ± 3.5 ) ( P ＜ 0. 05 ). Conclusions Transfection with PTEN may suppress the invasive and migration ability of ovarian cancer cell line A2780 depending on its phosphatase activity, and the suppressive effect may be due to the down-regulation of MMP-9 in the cancer cells.