摘要
目的:探讨乳腺癌细胞中上皮间质转化(epithelial-mesenchymal transition,EMT)与P-糖蛋白(P-glucoprotein,P-gp)介导的多药耐药(multidrug resistance,MDR)的关系及其可能的机制。方法:将携Snail基因的真核表达载体pcDNA-Snail转染人乳腺癌细胞MCF-7,用多柔比星(doxorubicin,DOX)诱导各组细胞耐药。免疫细胞荧光检测上皮标志物E-钙黏素(E-cadherin)、间质标志物波形蛋白(vimentin)以及P-gp的表达,MTT检测耐药细胞的增殖,RT-PCR检测Snail、MDR1、p38-MAPK mRNA的表达。结果:细胞免疫荧光显示,转染pcDNA-Snail载体后,MCF-7细胞发生EMT,E-cadherin表达显著降低,vimentin表达显著升高;P-gp在发生EMT的MCF-7细胞中表达显著升高。经DOX诱导,MCF-7/Snail细胞的耐药能力较MCF-7/DOX细胞显著增强(P<0.05)。RT-PCR显示,MCF-7细胞发生EMT后,p38-MAPK表达显著升高(P<0.05),MDR1表达较亲本细胞明显升高(P<0.01)。结论:MCF-7细胞发生EMT后,可能通过p38-MAPK引发P-gp介导的MDR。
Objective ：To investigate the relationship between epithelial-mesenchymal transition （EMT） and P-glucoprotein （P-gp）-induced muhidmg resistance （MDR） in breast cancer cells and the corresponding mechanisms. Methods： Eukaryotic expression vector pcDNA-Snail was constructed and then transfected into human breast cancer cell line MCF-7. Multidrug resistance was induced by doxorubicin （DOX） in different groups. Expressions of epithelial marker E-cadherin, interstitial marker vimentin, and P-glucoprotein （P-gp） were detected by immunofluorescence. MTT assay was used to measure the proliferation of drug resistant MCF-7 cells. Expressions of Snail, MDR1, and p38-MAPK mRNA were evaluated by RT-PCR. Results： Immunofluorescence showed that MCF-7 cells had EMT after transfection with pcD- NA-Snail vector. The expression of E-cadherin was downregnlated, and expressions of vimentin and P-gp were upregulated in EMT-like MCF-7 cells. Drug resistance of MCF-7/Snail cells was significantly enhanced compared with MCF-7 cells after induction by DOX （ P 〈 0.05 ）. The expressions of MDR1 and p38-MAPK mRNA in EMT-like cells were also significantly increased compared with those in parental MCF-7 cells （ P 〈 0.05 ）. Conclusion ： EMT may trigger DOX-induced and P-gp-mediated MDR via p38-MAPK in MCF-7 cells.
出处
《中国肿瘤生物治疗杂志》
CAS
CSCD
北大核心
2010年第2期144-148,共5页
Chinese Journal of Cancer Biotherapy
基金
山东省自然科学基金资助项目(No.Y2008C75)~~
关键词
SNAIL基因
乳腺肿瘤
上皮间质转化
多药耐药
P-糖蛋白
Snail gene
breast neoplasms
epithelial-mesenchymal transition （EMT）
muhidrug resistance （MDR）
P-glucoprotein （ P-gp ）
