溶酶体跨膜蛋白5在卵巢癌细胞上皮-间质转化中的作用

Lysosomal protein transmembrane 5 affects epithelial-mesenchymal transformation in ovarian cancer cells

目的 检测溶酶体跨膜蛋白5 (LAPTM5)在人卵巢腺癌细胞(OVCAR3)中的表达,观察LAPTM5对OVCAR3细胞迁移、侵袭能力的影响,并探讨LAPTM5在上皮-间质转化(EMT)过程中的作用。方法 采用Western blotting检测OVCAR3中LAPTM5的表达情况;将OVCAR3细胞随机分为2组,分别转染空载质粒和LAPTM5沉默质粒,实时定量PCR检测转染效率和EMT相关标志物E-cadherin、N-cadherin、Vimentin mRNA的表达水平。结果 LAPTM5在OVCAR3细胞中表达上调(P <0.01);与对照组相比,转染LAPTM5沉默质粒的OVCAR3细胞株迁移、侵袭能力明显下降(P <0.05),EMT上皮标志物E-cadherin mRNA表达水平显著上调(P <0.05),间质标志物N-cadherin mRNA和Vimentin mRNA表达下调(P <0.05)。结论 LAPTM5在人卵巢腺癌细胞OVCAR3中表达上调,LAPTM5可能通过EMT影响OVCAR3细胞迁移和侵袭的能力。

Objective To detect the expression of lysosomal protein transmembrane 5(LAPTM5) in human ovarian adenocarcinoma cells(OVCAR3),observe the effect of LAPTM5 on the migration and invasion of OVCAR3,and explore the role of LAPTM5 in epithelial-mesenchymal transformation(EMT). Methods Western blotting was used to detect LAPTM5 expression in OVCAR3. OVCAR3 cells were randomly divided into 2 groups and transfected with empty plasmids and LAPTM5 silent plasmids,respectively. Real-time quantitative polymerase chain reaction was used to measure transfection efficiency and mRNA expression levels of EMT-related markers,E-cadherin,N-cadherin,and Vimentin. Results LAPTM5 is expressed in the ovarian cancer cell line OVCAR3(P < 0.01). Compared to the control group,the migration and invasion ability of OVCAR3 cell lines transfected with LAPTM5 silenced plasmid were significantly decreased(P < 0.05),and the mRNA expression level of the EMT epithelial marker,E-cadherin,was significantly up-regulated(P < 0.05).The expression of N-cadherin mRNA and Vimentin mRNA in interstitial markers(P < 0.05) were down-regulated. Conclusion LAPTM5 is expressed in the ovarian cancer cell line OVCAR3,and LAPTM5 may affect the migration and invasion ability of OVCAR3 cells through EMT.