CAJ | 학술논문

目的研究不同转移能力的卵巢上皮性癌(卵巢癌)细胞中胰蛋白酶样Ⅱ型跨膜丝氨酸蛋白酶(matriptase)的表达,并探讨其与卵巢癌细胞转移能力的相关性.方法选取卵巢癌细胞系HO8910及其配对高转移能力细胞系HO8910PM,应用体外划痕实验及穿膜(transwell)小室法检测两种细胞的体外迁移及侵袭能力,应用逆转录(RT)-PCR和细胞免疫荧光技术检测两种细胞中matriptase mRNA和蛋白的表达水平,并对卵巢癌细胞中matriptase mRNA和蛋白的表达水平与其体外迁移、侵袭能力的相关性进行Pearson相关性分析.结果体外划痕实验显示,划痕后培养24 h,HO8910PM细胞的迁移距离为(347±8)μm,明显大于HO8910细胞的(154±10) μm(P ＜0.01)；穿膜小室法检测显示,24 h后HO8910PM细胞的穿膜细胞数为(90.7±2.1)个,明显高于HO8910细胞的(63.3±1.5)个(P＜0.01).RT-PCR技术检测显示,HO8910PM细胞中matriptase mRNA的表达水平为0.72±0.03,明显高于HO8910细胞的0.38±0.04(P ＜0.01)；细胞免疫荧光技术检测显示,HO8910PM细胞中matriptase蛋白的表达水平为15.6±0.8,明显高于HO8910细胞的7.6±1.3(P＜0.01).Pearson相关性分析显示,卵巢癌细胞中matriptase mRNA和蛋白的表达水平与其迁移(r=0.992,r =0.971;P ＜0.01)及侵袭(r=0.973,r =0.958;P ＜0.01)能力均呈明显正相关.结论高转移能力的卵巢癌细胞系HO8910PM细胞中matriptase mRNA和蛋白的表达水平均明显高于低转移能力的卵巢癌细胞系HO8910细胞,matriptase基因的表达与卵巢癌细胞的迁移和侵袭能力可能存在相关性,其机制有待进一步研究.
목적 연구불동전이능력적란소상피성암(란소암)세포중이단백매양Ⅱ형과막사안산단백매(matriptase)적표체,병탐토기여란소암세포전이능력적상관성.방법 선취란소암세포계HO8910급기배대고전이능력세포계HO8910PM,응용체외화흔실험급천막(transwell)소실법검측량충세포적체외천이급침습능력,응용역전록(RT)-PCR화세포면역형광기술검측량충세포중matriptase mRNA화단백적표체수평,병대란소암세포중matriptase mRNA화단백적표체수평여기체외천이、침습능력적상관성진행Pearson상관성분석.결과 체외화흔실험현시,화흔후배양24 h,HO8910PM세포적천이거리위(347±8)μm,명현대우HO8910세포적(154±10) μm(P ＜0.01)；천막소실법검측현시,24 h후HO8910PM세포적천막세포수위(90.7±2.1)개,명현고우HO8910세포적(63.3±1.5)개(P＜0.01).RT-PCR기술검측현시,HO8910PM세포중matriptase mRNA적표체수평위0.72±0.03,명현고우HO8910세포적0.38±0.04(P ＜0.01)；세포면역형광기술검측현시,HO8910PM세포중matriptase단백적표체수평위15.6±0.8,명현고우HO8910세포적7.6±1.3(P＜0.01).Pearson상관성분석현시,란소암세포중matriptase mRNA화단백적표체수평여기천이(r=0.992,r =0.971;P ＜0.01)급침습(r=0.973,r =0.958;P ＜0.01)능력균정명현정상관.결론 고전이능력적란소암세포계HO8910PM세포중matriptase mRNA화단백적표체수평균명현고우저전이능력적란소암세포계HO8910세포,matriptase기인적표체여란소암세포적천이화침습능력가능존재상관성,기궤제유대진일보연구.
Objective To study the expression and significance of matriptase in different metastatic potential of human ovarian cancer cells.Methods High-metastatic human ovarian cancer cell HO8910PM and ovarian cancer cell HO8910 were collected.The ability of metastatic of the former was stronger than that of the latter.Compared the ability of invasion and migration in HO8910PM and HO8910 by scratch assay and by millicell chamber artificial reconstituted basement membrane invasion assay.Detected the matriptase mRNA and protein expression levels in HO8910PM and HO8910 through reverse transcription(RT)-PCR and immunocytochemistry methods.Results The 24 hours' migration distance(347 ± 8) μm of HO8910PM cells were significantly higher than that in HO8910 group (154 ± 10) μm (P ＜ 0.01) ;The number of HO8910PM cells that penetrated the matrigel after 24 hours' incubation were significantly higher than that in HO8910 group (90.7 ±2.1 vs 63.3 ± 1.5,P ＜0.01).The expression of matriptase mRNA in HO8910PM cells was higher than that in HO8910 group (0.72 ± 0.03 vs 0.38 ± 0.04,P ＜ 0.01).The migration was positively correlated with the matriptase mRNA expression levels (r =0.992,P ＜ 0.01); and the invasiveness was also positively correlated with the matriptase mRNA expression levels (r =0.973,P ＜0.01).As far protein level,the expression of matriptase protein in HO8910PM cells was higher than that in HO8910 group (15.6 ±0.8 vs 7.6 ± 1.3,P ＜0.01).The migration was positively correlated with matriptase protein expression levels (r =0.971,P ＜ 0.01) ;And the invasiveness was also positively correlated with the matriptase protein expression levels (r =0.958,P ＜ 0.01).Conclusions The relationship between the expression levels of matriptase and the metastatic of ovarian cancer cells may be correlative.The function of matriptase in ovarian cancer cells metastatic machanism still need to be confirmed.