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目的:利用实验室构建的微流控芯片对乳腺癌细胞(MDA-MB-231)进行捕获,提高捕获率并保证细胞活性,实现再培养。抗肿瘤药物阿霉素处理正常培养和再培养的细胞,分析细胞内的基因表达变化。方法:对微流控芯片进行基底修饰,利用MUC1抗原与抗体特异性结合捕获肿瘤细胞,优化捕获条件提高捕获率。对微流控芯片捕获的细胞进行分离、收集和再培养。用1μmol/L阿霉素对正常培养和再培养的细胞分别孵育24h,然后提取RNA并逆转录合成c DNA。选择乳腺癌细胞中高表达及与肿瘤转移相关的基因FN1、ITGA6和LAMB3设计引物,以c DNA为模板分别进行RT-PCR扩增,对琼脂糖凝胶电泳结果进行灰度分析。结果:经MUC1抗体修饰的微流控芯片能有效地捕获肿瘤细胞,捕获率达80%±3%,释放率约98%,细胞释放后存活率高实现再培养。阿霉素对正常培养和再培养的乳腺癌细胞中FN1、ITGA6和LAMB3的基因表达均有抑制作用。结论:MUC1抗体修饰的微流控芯片能有效捕获乳腺癌细胞并实现再培养,捕获前后细胞内基因表达无显著差异,均能产生药物敏感性。
OBJECTIVE: To capture the breast cancer cells (MDA-MB-231) by using the microfluidic chip constructed in laboratory and to improve the capture rate and cell viability and achieve the re-culture. The antitumor drug doxorubicin treated normal and re-cultured cells and analyzed the changes of intracellular gene expression. Methods: The microfluidic chip was modified by substrate. The specific binding of MUC1 antigen and antibody was used to capture tumor cells. The capture conditions were optimized to improve the capture rate. The cells captured by the microfluidic chip are isolated, collected and re-cultured. The normal cultured and the cultured cells were respectively incubated with 1 μmol / L doxorubicin for 24 h, then RNA was extracted and cDNA was reverse transcribed into c DNA. Select the high expression of breast cancer cells and tumor metastasis-related genes FN1, ITGA6 and LAMB3 primers designed to c DNA template were RT-PCR amplification, agarose gel electrophoresis results were grayscale analysis. Results: The MUC1 antibody-modified microfluidic chip could effectively capture tumor cells with the capture rate of 80% ± 3% and the release rate of about 98%. The cell survival rate after re-culturing was high. Doxorubicin has an inhibitory effect on the gene expression of FN1, ITGA6 and LAMB3 in normal and re-cultured breast cancer cells. Conclusion: MUC1 antibody-modified microfluidic chip can effectively capture breast cancer cells and achieve re-culture, before and after the capture of gene expression was no significant difference, can produce drug sensitivity.