磁性分选肺腺癌始动细胞的异常miRNAs验证  

Aberrant miRNA validation in lung adenocarcinoma initiating cells spared by magnetic bead from A549

作　　者：张振华[1] 杨红茹[1] 周杰[1] 邓曦[1] 吴敬波[1] 林盛[1] 

机构地区：[1]泸州医学院附属医院肿瘤科,四川省646000

出　　处：《中华临床医师杂志（电子版）》2013年第20期74-76,共3页Chinese Journal of Clinicians(Electronic Edition)

基　　金：国家自然科学基金(81201682)

摘　　要：目的利用磁性活细胞分选法(magnetic activated cell sorting,MACS)从人A549肺腺癌细胞中分离得到CD133+标记细胞,通过CD133/CD326双阳性检测探讨分离效果,初步分析差异表达miRNAs对该亚群细胞的调控功能。方法将对数生长期的A549细胞离心收集,重悬于无血清培养基中,培养至第二代后利用CD133磁珠标记后分选,流式细胞术及免疫荧光验证分选后细胞的CD133/CD326双阳性率;结合前期实验miRNA芯片结果,挑选兴趣分子进行定量PCR验证。结果利用CD133磁珠分选得到的阳性细胞亚群高表达CD133/CD326分子,结合前期miRNA芯片结果,选出在CD133+/CD326+细胞亚群中表达上调的miR-663,miR-183,miR-125a-5p,miR-127,miR-520h及表达下调的miR-18b,miR-29ab,miR-17和miR-155行定量PCR检测证实miR-29ab,miR-155,miR-183,miR-127-3p及miR-17的表达趋势与芯片结果相符。结论利用磁珠分选方式能获得CD133+/CD326+高表达肺腺癌始动细胞亚群且包括miR-183等在内的6条分子与芯片结果一致,可能在肺腺癌始动细胞生物学行为的调控中发挥重要作用。Objective To validate magnetic activated cell sorting （MACS） is another means in enriching lung adenocarcinoma initiating cells from normal A549 cells and based on quantitative RT-PCR to analyze regulatory roles of this subpopulation. Methods After obtaining the lung adenocarcinoma initiating cells by MACS, we utilize flow cytometry analysis and imrnunofluoreseence to verify CD 133/CD326 expression of this subpopulation and choose 10 miRNAs to perform quantitative RT-PCR. Results We obtained CD133/CD326 high expression subpopulation by MACS. 10 miRNAs chose for quantitative RT-PCR and 6 miRNAs expression trend were consist with army data including miR-29ab, miR-155, miR-183, miR-127-3p, miR-17. Conclusion MACS can enrich CD133＋/CD326＋ subpopulation and 6 miRNAs expression trend were consist with array data, which may play important roles in regulating the biobehavior of lung adenocarcinoma initiating cells.

关 键 词：微RNAS 磁性活细胞分选 肿瘤始动细胞 

分 类 号：R734.2[医药卫生—肿瘤]