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目的:比较人乳腺癌亲代敏感株MCF-7和多耐药细胞株MCF-7/A中ALDH1,P-gp及CD44v6的表达差异,探讨多耐药与侵袭转移作用机制。方法:以阿霉素低浓度加量持续诱导法,建立人乳腺癌多耐药细胞模型株MCF-7/A,用ALDEFLUOR分析法检测乳腺癌干细胞功能性标志物ALDH1阳性细胞在亲代敏感株MCF-7与多耐药细胞株MCF-7/A中所占比例,Transwell侵袭实验论证乳腺癌胞耐药是否伴随相应侵袭转移能力的改变,免疫细胞化学染色及荧光定量PCR分析亲代敏感株MCF-7与多耐药细胞株MCF-7/A中MDR1(P-gp)和CD44V6的表达变化。结果:耐药细胞株MCF-7/A较亲代敏感株ALDH1阳性细胞比例数明显增高;耐药细胞株MCF-7/A穿膜细胞数明显多于敏感细胞株MCF-7;耐药细胞株MCF-7/A中P-gp和CD44v6表达均明显高于亲代敏感细胞株。结论:乳腺癌的多耐药不仅包含获得性耐药细胞比例增加,同时伴有肿瘤干细胞的富集造成的内在性耐药增加,耐药的乳腺癌细胞伴有侵袭能力增强。
OBJECTIVE: To compare the expression of ALDH1, P-gp and CD44v6 in human breast cancer parental susceptible strain MCF-7 and multi-drug resistant cell line MCF-7 / A, and to explore the mechanisms of multidrug resistance and invasion and metastasis. METHODS: Human multidrug-resistant human breast cancer cell line MCF-7 / A was established by sustained induction with low concentration of doxorubicin. The ALDEFLUOR assay was used to detect the expression of ALDH1-positive cells, a functional marker of breast cancer stem cells, -7 and multi-drug resistant cell line MCF-7 / A. Transwell invasion assay demonstrated that breast cancer cell resistance was accompanied by the corresponding invasion and metastasis. Immunocytochemical staining and real-time PCR analysis of the parental susceptible strain MCF- 7 and MDR1 (P-gp) and CD44V6 in multi-drug resistant cell line MCF-7 / A. Results: The number of MCF-7 / A-resistant cells was significantly higher than that of the parentally-susceptible strain ALDH1-positive cells. The number of transmembrane cells in MCF-7 / A cell line was significantly higher than that in MCF-7 cell line The expressions of P-gp and CD44v6 in MCF-7 / A were significantly higher than those of parental sensitive cell lines. Conclusion: Multidrug resistance of breast cancer not only includes the increase of the proportion of acquired drug-resistant cells, but also the increase of intrinsic resistance caused by the enrichment of cancer stem cells, and the enhancement of invasion ability of resistant breast cancer cells.