The miR-15/107 family comprises a group of 10 paralogous microRNAs （miRNAs）,sharing a 5＇ AGCAGC sequence.These miRNAs have overlapping targets.In order to characterize the expression of miR-15/107 family miRNAs,we ...The miR-15/107 family comprises a group of 10 paralogous microRNAs （miRNAs）,sharing a 5＇ AGCAGC sequence.These miRNAs have overlapping targets.In order to characterize the expression of miR-15/107 family miRNAs,we employed customized TaqMan Low-Density micro-fluid PCR-array to investigate the expression of miR-15/107 family members,and other selected miRNAs,in 11 human tissues obtained at autopsy including the cerebral cortex,frontal cortex,primary visual cortex,thalamus,heart,lung,liver,kidney,spleen,stomach and skeletal muscle.miR-103,miR-195 and miR-497 were expressed at similar levels across various tissues,whereas miR-107 is enriched in brain samples.We also examined the expression patterns of evolutionarily conserved miR-15/107 miRNAs in three distinct primary rat brain cell preparations （enriched for cortical neurons,astrocytes and microglia,respectively）.In primary cultures of rat brain cells,several members of the miR-15/107 family are enriched in neurons compared to other cell types in the central nervous system （CNS）.In addition to mature miRNAs,we also examined the expression of precursors （pri-miRNAs）.Our data suggested a generally poor correlation between the expression of mature miRNAs and their precursors.In summary,we provide a detailed study of the tissue and cell type-specific expression profile of this highly expressed and phylogenetically conserved family of miRNA genes.展开更多
[目的]探讨miR-15a在多发性骨髓瘤(MM)患者外泌体中的表达及外泌体对肿瘤发生发展的意义。[方法]选择在我院治疗的MM患者71例及健康志愿者20例,分析miR-15a在血浆外泌体中的表达水平;并以人源MM细胞株MM.1R细胞为实验模型,采用增殖实验...[目的]探讨miR-15a在多发性骨髓瘤(MM)患者外泌体中的表达及外泌体对肿瘤发生发展的意义。[方法]选择在我院治疗的MM患者71例及健康志愿者20例,分析miR-15a在血浆外泌体中的表达水平;并以人源MM细胞株MM.1R细胞为实验模型,采用增殖实验观察外泌体对肿瘤细胞增殖的影响,采用流式细胞术检测肿瘤细胞凋亡,采用Western blot检测凋亡通路相关蛋白的表达。[结果]与健康志愿者相比,MM患者的血浆miR-15a水平显著降低(4.45±0.20 vs 12.86±0.63,P<0.001),且与患者临床分期有关(P=0.023)。当使用MM患者来源的外泌体处理MM.1R细胞后,实验组细胞增殖率较对照组提高,在12h、24h、48h时分别提高了14.1%、34.0%、52.7%(P<0.01);而健康志愿者来源的外泌体则使细胞增殖率降低,在12h、24h、48h时分别降低了12.4%、20.8%、27.5%(P<0.01)。流式细胞术结果显示,MM患者来源外泌体抑制了肿瘤细胞凋亡,24h凋亡率减少了3.3%(P=0.005);而健康志愿者来源外泌体促进了肿瘤细胞凋亡,24h凋亡率增加了7.8%(P<0.001)。Western blot检测发现MM组的Bcl-2表达增加,Bax和caspase-3表达下降,与健康志愿者来源的外泌体作用相反。[结论]促凋亡因子miR-15a在MM患者外泌体中的表达水平低于健康人群,且MM患者来源的外泌体具有促进肿瘤发展的作用。展开更多
基金supported by the National Institutes of Health,USA(Grant Nos.AG042419,NS085830 and AG028 383)
文摘The miR-15/107 family comprises a group of 10 paralogous microRNAs （miRNAs）,sharing a 5＇ AGCAGC sequence.These miRNAs have overlapping targets.In order to characterize the expression of miR-15/107 family miRNAs,we employed customized TaqMan Low-Density micro-fluid PCR-array to investigate the expression of miR-15/107 family members,and other selected miRNAs,in 11 human tissues obtained at autopsy including the cerebral cortex,frontal cortex,primary visual cortex,thalamus,heart,lung,liver,kidney,spleen,stomach and skeletal muscle.miR-103,miR-195 and miR-497 were expressed at similar levels across various tissues,whereas miR-107 is enriched in brain samples.We also examined the expression patterns of evolutionarily conserved miR-15/107 miRNAs in three distinct primary rat brain cell preparations （enriched for cortical neurons,astrocytes and microglia,respectively）.In primary cultures of rat brain cells,several members of the miR-15/107 family are enriched in neurons compared to other cell types in the central nervous system （CNS）.In addition to mature miRNAs,we also examined the expression of precursors （pri-miRNAs）.Our data suggested a generally poor correlation between the expression of mature miRNAs and their precursors.In summary,we provide a detailed study of the tissue and cell type-specific expression profile of this highly expressed and phylogenetically conserved family of miRNA genes.
文摘[目的]探讨miR-15a在多发性骨髓瘤(MM)患者外泌体中的表达及外泌体对肿瘤发生发展的意义。[方法]选择在我院治疗的MM患者71例及健康志愿者20例,分析miR-15a在血浆外泌体中的表达水平;并以人源MM细胞株MM.1R细胞为实验模型,采用增殖实验观察外泌体对肿瘤细胞增殖的影响,采用流式细胞术检测肿瘤细胞凋亡,采用Western blot检测凋亡通路相关蛋白的表达。[结果]与健康志愿者相比,MM患者的血浆miR-15a水平显著降低(4.45±0.20 vs 12.86±0.63,P<0.001),且与患者临床分期有关(P=0.023)。当使用MM患者来源的外泌体处理MM.1R细胞后,实验组细胞增殖率较对照组提高,在12h、24h、48h时分别提高了14.1%、34.0%、52.7%(P<0.01);而健康志愿者来源的外泌体则使细胞增殖率降低,在12h、24h、48h时分别降低了12.4%、20.8%、27.5%(P<0.01)。流式细胞术结果显示,MM患者来源外泌体抑制了肿瘤细胞凋亡,24h凋亡率减少了3.3%(P=0.005);而健康志愿者来源外泌体促进了肿瘤细胞凋亡,24h凋亡率增加了7.8%(P<0.001)。Western blot检测发现MM组的Bcl-2表达增加,Bax和caspase-3表达下降,与健康志愿者来源的外泌体作用相反。[结论]促凋亡因子miR-15a在MM患者外泌体中的表达水平低于健康人群,且MM患者来源的外泌体具有促进肿瘤发展的作用。
