摘要
目的:探讨电压门控钠离子通道(voltage-gated sodium channels,VGSCs)在宫颈癌细胞中的表达、分布及其对该细胞增殖、侵袭能力的影响。方法:用Western blot了解VGSCs在不同宫颈癌细胞株中表达水平,采用免疫荧光检测VGSCs蛋白在细胞中的定位。分别用MTT法和Matrigel法检测VGSCs对宫颈癌细胞的增生和侵袭作用。以半定量RT-PCR和RNAi方法确认宫颈癌细胞中VGSC表达亚型及其功能。结果:在检测的4种宫颈癌细胞株中,ME180细胞表达较高水平VGSCs蛋白,其存在于细胞膜和细胞质中。VGSCs抑制剂-河豚毒素(TTX)对ME180细胞增生无影响(P>0.05),但呈浓度依赖性抑制细胞的Matrigel侵袭(P<0.05)。在ME180细胞中检测到Nav 1.2、Nav 1.6和Nav 1.7三种VGSCs亚型,其中Nav 1.6 mRNA占总mRNA的78%,RNAi抑制Nav 1.6mRNA表达后可降低细胞侵袭达52%(P<0.05)。结论:VCSCs在宫颈癌ME180细胞株高表达,Nav 1.6为其主要表达亚型,增加了体外癌细胞的侵袭转移。
Objective： The present study investigates the expression and distribution of voltageated sodium channels （ VGSCs ）, as well as its effects on the proliferation and invasion of human cervical cancer cells. Methods： Western blot analysis and immunofluorescence were used to detect the level and intracellular distribution of the VGSC protein in cervical cancer cell lines, respectively. Methyl thiazolyl tetrazolium assay and Matrigel methods tested the effect of VGSCs on the proliferation and invasion of the cervical cancer ME 180 cells, respectively. The expression of the VGSC subtype in ME 180 cells was determined using semi-quantitative polymerase chain reaction and ribonucleic acid interference （ RNAi ）. Results： Cervical cancer ME180 cells expressed the highest level of VGSC protein in the four cervical cancer cell lines tested. VGSC protein was primarily found in the plasma membrane and cytoplasm of the cells. Tetrodotoxin, an inhibitor of VGSCs, had no effect on the proliferation of the ME 180 cells, but decreased Matrigel invasion in a dose-dependent manner （ P 〈 0.05 ）. Three VGSC isoforms （ Navl.2, Nav1.6, and Navl.7 ） were identified to be expressed in the ME180 cells, among which Navl.6 mRNA was at the highest level （ P 〈 0.05 ）. RNAi targeting Navl.6 mRNA decreased the invasion of the cells by 52% （ P 〈 0.05 ）. Conclusion： VGSC subtype Nay1.6 was found to be expressed predominantly in human cervical cancer ME180 cells. The expression of Navl.6 in the cells increases the invasion of the cells in vitro.
出处
《中国肿瘤临床》
CAS
CSCD
北大核心
2012年第4期189-193,共5页
Chinese Journal of Clinical Oncology
基金
教育部留学回国人员科研启动基金
北京市自然科学基金(编号:7112052)资助~~
