摘要
目的:探讨小窝蛋白-1(caveolin-1,CAV-1)的表达与绒毛膜癌高侵袭力之间的相关性,并了解RNA干扰沉默CAV-1基因对高侵袭力绒毛膜癌JEG-3细胞侵袭力的抑制作用。方法:(1)应用Matrigel体外侵袭试验和噻唑盐比色实验[3-(4,5)-dimethylthiahiazo(-z-y1)-3,5-di-phenytetrazoliumro-mide,MTT]检测绒毛膜癌JEG-3细胞和JAR细胞之间侵袭能力和增殖能力的差异;(2)应用半定量逆转录聚合酶链反应(reverse transcriptase polymerase chain reaction,RT-PCR)方法检测CAV-1mRNA在人正常早孕绒毛组织、不同侵袭力人绒毛膜癌细胞株JEG-3及JAR表达的差异;(3)将CAV-1小干扰RNA(small interfering RNA,siRNA)转染至高侵袭力绒毛膜癌JEG-3细胞后,采用RT-PCR检测转染后JEG-3细胞CAV-1mRNA表达的变化,并应用Matrigel体外侵袭试验和MTT法检测JEG-3细胞侵袭和增殖能力的变化。结果:(1)JEG-3细胞的侵袭能力显著高于JAR细胞(P<0.05),但JEG-3和JAR细胞之间增殖能力的差异无统计学意义(P>0.05);(2)RT-PCR结果显示在人正常早孕绒毛组织、高侵袭性JEG-3细胞以及低侵袭性JAR细胞中均可检测到CAV-1mRNA,将上述三者中CAV-1mRNA的表达量进行两两比较,发现人绒毛膜癌细胞株中CAV-1mRNA的表达明显高于人正常早孕绒毛组织,而具有高侵袭性JEG-3细胞中CAV-1mRNA的表达明显高于JAR细胞,差异具有统计学意义(P<0.05),相关性分析显示CAV-1表达水平与绒毛膜癌细胞侵袭力呈正相关(r=0.86,P<0.05);(3)CAV-1 siRNA可有效抑制CAV-1mRNA的表达,并削弱绒毛膜癌细胞的侵袭和增殖能力。结论:CAV-1基因表达可显著增强绒毛膜癌细胞的侵袭潜能,CAV-1 siRNA可抑制绒毛膜癌细胞的侵袭和增殖能力。
Objective To investigate the association between the expression of caveolin-1 ( CAV-1 ) and the invasion of choriocarcinoma, and to explore the effect of CAV-1 small interfering RNA ( siRNA ) on the invasion of choriocarcinoma cell line JEG-3. Methods ( 1 ) Matrigel invasion assay and 3- ( 4,4 ) -dimethylthiahiazo ( -z-yl ) -3,5 -di-phenytetrazoliumormide ( MTr ) assay were used to examine the difference in invasion and proliferation ability between JEG-3 cells and JAR cells; (2) Expression of caveolin-1 gene in the human chorionic villi tissues and chorionicnoma cell lines (JEG-3 cells and JAR cells ) were detected by semi-quantitative RT-PCR. (3) The effect of CAV-1 siRNA transfection on the expression of CAV-1 mRNA, and the invasion and proliferation ability of JEG-3 cells were measured by RT-PCR, Matrigel invasion assay and MTT assay. Results ( 1 ) The invasion ability of JEG-3 cell line was stronger than that of JAR cell line (P 〈0.05 ) , but the difference in proliferation ability between JAR and JEG-3 was not obvious ( P 〉 0.05 ) ; ( 2 ) The expression of caveolin-1 gene in chorionicnoma cell lines was significantly stronger than that in the human normal chorion( P 〈0.05 ) , and the expression of caveolin-1 gene in JEG-3 cells was stronger than that in the JAR cells ( P 〈 0.05 ). The data suggested that there was significantly positive correlation between caveolin-1 and the invasiveness of chorionicnoma cells ( r = 0. 086, P 〈 0.05 ) ; (3) CAV-1 siRNA could knock-out the expression of CAV-1 mRNA, and inhibit the invasion and proliferation ability of chorionicnoma cells. Conclusion CAV-1 can promote the invasion ability of chorionicnoma cells. CAV-1 siRNA can inhibit the invasion and proliferation ability of chorionicnoma cells.
出处
《中南大学学报(医学版)》
CAS
CSCD
北大核心
2008年第4期331-337,共7页
Journal of Central South University :Medical Science
基金
湖南省科技计划项目(05FJ3077)~~