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作 者:秦洁[1,2] 李屹[3] 刘永琦[1,4,5] 王倩[1] 舍雅莉[1,4] 骆亚莉[1,4]
机构地区:[1]甘肃中医学院系统生物学与中医药转化研究所 [2]兰州大学校医院 [3]兰州大学基础医学院遗传学研究所 [4]甘肃中医学院省中药药理与毒理学重点实验室中西医结合基础室 [5]甘肃中医学院敦煌医学与转化省部共建教育部重点实验室,甘肃兰州730020
出 处:《基础医学与临床》2013年第9期1112-1117,共6页Basic and Clinical Medicine
基 金:国家自然科学基金(81060351;H2810);甘肃省教育厅科研基金(0906-03)
摘 要:目的探讨人骨髓间充质干细胞(HMSC-bm)在肺腺癌微环境中遗传稳定性的变化。方法通过6孔板结合Transwell小室建立HMSC-bm和肺腺癌A549细胞的共培养体系,倒置相差显微镜下观察细胞的形态;MTT法检测细胞的增殖能力;常规染色体及G显带核型分析细胞的染色体;Western blot检测组蛋白去乙酰化酶4(HDAC4)的表达。以单独培养的HMSC-bm,A549作为对照组。结果共培养组细胞胞核大而深染,可见病理性核分裂象,HMSC-bm组无明显变化;细胞增殖曲线呈S型,共培养组细胞于第4天开始增殖速率快于HMSC-bm组;共培养组染色体数目为亚三倍体、三倍体,有明显的染色体异常;共培养组细胞HDAC4表达明显高于HMSC-bm组(P<0.01)。结论肺腺癌微环境可改变HMSC-bm的增殖速率和其遗传稳定性。Objective To investigate the genetic stability of human bone marrow mesenchymal stem cells ( HMSC- bm) under pulmonary adenocarcinoma microenvironment. Methods A co-cultured system of HMSC-bm and A549 cells was established through the combination of 6 well culture plate and Transwell chamber as experimental group while HMSC-bm and A549 cells were cultured as the control groups. The morphology of the cells was observed by phase-contrast microscopy. The proliferation curve of mesenchymal stem cells was tested by MTT. Chromosome was examined by karyotyping analysis. The expression of HDAC4 protein was detected by Western blot. Results Cells in the co-culture group showed that the cell nucleus became bigger than the nucleus in HMSC-bm group and exhibiIed anachromasis in 7 days. The proliferation curve showed faster proliferation of co-cuhured HMSC-hm than HMSC-bm. Karyotyping analysis showed that the chromosome number of co-cultured HMSC-bm was hypotriploid and triploid and the chromosome was grossly abnormal. The expression of HDAC4 protein in co-cultured HMSC-bm significantly increased compared to that in HMSC-bm (P 〈0. 01 ). Conclusion Pulmonary adenocareinoma microenvironment may influence the proliferation speed and the genetic stability of HMSC-bm.
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