耐顺铂卵巢癌患者腹水中ALDH1阳性肿瘤细胞的生物学特性

Biological characteristics of ALDH1 positive tumor cells in CDDP resistance ovarian cancer patients

目的:探讨卵巢癌顺铂耐药患者腹水中ALDH1阳性肿瘤细胞的生物学特征。方法:从卵巢癌顺铂耐药患者腹水中分离癌细胞,流式分选法得到ALDH1阳性和ALDH1阴性细胞,荧光定量RT-PCR检测两组细胞的干细胞标志CD44、CD90、CD133及上皮间质化相关标志E-cadherin、N-cadherin、Vimentin及Fibronectin在mRNA水平的表达,MTT法测定细胞增殖曲线和对顺铂的耐药指数,Transwell小室侵袭实验、克隆形成实验、悬浮成球实验分别观察细胞侵袭能力、增殖分化能力及干细胞的潜能。结果:与ALDH1阴性细胞相比,ALDH1阳性细胞的干细胞标志CD44、CD90、CD133表达升高,上皮化标志E-cadherin降低,间质化标志N-cadherin、Vimentin表达明显升高。MTT测定生长曲线显示ALDH1阳性细胞增殖明显增快,对顺铂的耐药指数为14.2,ALDH1阳性细胞的穿膜细胞数、克隆形成数以及成球数目显著高于ALDH1阴性细胞(P<0.05)。结论:ALDH1阳性细胞具备干细胞特征,其耐药性和侵袭能力增强。

Objective：To explore the biological characteristics of ALDH＋cells from malignant ascites of CDDP resistance ovarian cancer.Methods：Ovarian cancer cells were isolated from the malignant ascites of CDDP resistance ovarian cancer patients to separate ALDH＋and ALDH-cells using flow cytometry.Real-time quantitative RT-PCR was used to detect the mRNA expressions of stem cell related genes CD44,CD90,CD133 and epithelial to mesenchymal transition related genes.The drug resistance and growth curve of cells were detected by MTT assay.Cell invasion,cell proliferation and differentiation,stem cell potential were observed by Transwell invasion assay,clone formation assay,suspension sphere forming assay.Results：The expression of E-cadherin was significantly decreased in the ALDH＋cells（ P〈0.05）,but the expressions of CD44,CD90,CD133,N-cadherin and Vimentin were significantly higher in the ALDH1＋cells.The drug resistant index of ALDH1＋cell kept on 14.2 times.The number of invasion cells,cell clones and suspended spheres were significantly higher in the ALDH1＋cells than the ALDH1-cells（ P〈0.05）.Conclusion：ALDH1＋cells have higher abilities of invasion,proliferation and potential of stem cells.