摘要
目的探讨低剂量辐射预处理对人骨髓间充质干细胞(human bone mesenchymal stem cells,hBMSCs)sur-vivin mRNA表达的影响。方法密度梯度离心法分离培养正常人骨髓间充质干细胞,流式细胞仪检测免疫表型,体外定向诱导hBMSC分化为脂肪细胞,特异性染色鉴定其分化能力;随机分为未予照射的对照组,以及5、10、20cGy照射的实验组,MTT法检测hBMSCs在不同剂量照射后细胞增殖情况;RT-PCR检测5、20cGy照射后细胞survivin mRNA表达。结果体外成功分离hBMSCs,流式细胞仪分析细胞表面抗原CD44和CD29强阳性,HLA-ABC弱阳性,CD34、CD45和HLA-DR阴性;hBMSCs在特定条件下可诱导分化成脂肪细胞;5、10、20cGy照射处理后细胞增殖能力明显增强,均高于对照组(P均<0.01);10cGy辐射剂量处理后48、72h较24h细胞增殖能力明显增强(P均<0.01),处理后72h较48h细胞增殖能力明显增强(P<0.01),差异具有统计学意义;5cGy和20cGy辐射后细胞的survivin mRNA表达上调,与对照组相比(P均<0.01),差异具有统计学意义。结论低剂量辐射可促进hBMSCs的增殖及survivin mRNA的表达。
Objective To investigate the effect of low dose radiation on the expression of survivin gene in human bone marrow mesenchymal stem cells(hBMSCs).Methods hBMSCs from healthy donors were isolated and expanded by Fircoll density gradient centrifugation;The adipogenic differentiation was tested and evaluated by specific staining methods.The proliferation of cells under different doses of radiation and different time were detected with MTT assay;The survivin gene mRNA was detected by the RT-PCR method.Results The expressions of CD44,CD71 and HLA-ABC were positive while the expressions of CD34,CD45,HLA-DR in hBMSCs were negative;The hBMSCs were successfully induced into adipogenic lineages and the proliferative ability of hBMSCs exposed to LDR was obviously enhanced as compared with control group(P all<0.01);the proliferation ability of hBMSCs were higher which exposed to 10cGy radiation with 48h and 72h than those were exposed to 10cGy radiation with 24h(P all<0.01),and the hBMSCs proliferation was significantly increased which were exposed to 10cGy radiation with 72h compared with 48h(P<0.01);The expression of the survivin mRNA was higher in the hBMSCs which exposed to 5cGy and 20cGy radiation than that of the control group(P all<0.01).Conclusion LDR promotes the proliferation and the survivin mRNA expression of hBMSCs.
出处
《宁夏医科大学学报》
2013年第5期497-501,476,共6页
Journal of Ningxia Medical University
基金
甘肃省自然科学基金暨中青年科技基金(3ZS051-A-25-106)
甘肃省科技重大专项(1102FKDA005)
