微小核糖核酸-204和211在人胚胎干细胞向视网膜色素上皮细胞分化过程的变化  

Expression of miR-204 and 211 in human embryonic stem cells differentiated into retinal pigment epithelial cells

作　　者：李文博[1] 李筱荣[1] 鲁振宇[2] 张云山[2] 东莉洁[1] 王飞[1] 董荣[2] 

机构地区：[1]天津医科大学眼科医院眼视光学院,300384 [2]天津市中心妇产科医院生殖医学中心

出　　处：《中华眼底病杂志》2013年第1期72-75,共4页Chinese Journal of Ocular Fundus Diseases

基　　金：国家自然科学基金（30973255）

摘　　要：目的观察人胚胎干细胞（hESC）向视网膜色素上皮（RPE）细胞诱导分化过程中微小核糖核酸（miRNA）204和211的变化特征。方法采用广1然分化法诱导hESC向RPE细胞分化，细胞鉴定。按细胞诱导分化HIf间秩序，分为hESC组、含色素细胞组、hESC源性RPE细胞组和原代培养的人胎RPE（hfRPE）细胞组。行miRNA基闪表达谱芯片分析、miRNA-204和211实时荧光逆转录聚合酶链反应（RTPCR）检测。结果miRNA芯片分析结果显示，随细胞诱导分化过程，miRNA204持续上调。其中，含色素细胞组足hESC组的5．026倍，hESC源性RPE细胞组是含色素细胞组的3．337倍；hfRPE细胞组是hESC源件RPE细胞的13．574倍。miRNA211在细胞诱导分化过程中上调不明显，但从hESC源性RPE细胞到hfRPE细胞，上调倍数为44．333倍，是miRNA表达谱中差异最大的miRNA。RTPCR检测结果显示，hESC组、含色素细胞组、hESC源性RPE细胞组、hfRPE细胞组miR-204相埘表达量分别为91．81±4．43、2263．09±206．39、5996．80±235．42、171676．45±999．82，差异有统计学意义（t=18．22、20．66、279．38，P〈0．001）；miRNA-211相对表达量分别为2．23±0．31、129．33±3．75、125．76±4．78、16682．00±352．97。含色素细胞组和hESC细胞组、hfRPE细胞组和hESC源性RPE细胞组miR-211相对表达量比较，差异均有统计学意义（t=58．58、81．24，P〈0．001）。结论miRNA-204和211在hESC向RPE细胞诱导分化过程中持续单一变化。Objective To observe the expression of miR-204 and 211 human embryonic stem cells （hESCs） differentiated into retinal pigment epithelial （RPE） ceils. Methods RPE cells were derived from hESCs hy natural differentiation method, and were identified, miRNA expression profiles and real time polymerase chain reaction （RT PCR） of miR-204 and 211 were generated from the following groups： hESCs, hESCs derived cells containing pigmented loci, hESCs derived RPE cells and human fetal RPE （hfRPE） cells. Results miRNA 204 was continuously up-regulated throughout the entire differentiation process of hESCs to RPE cells. It increased 5. 026 times in hESCs derived cells containing pigmented foci compared to hfRPE cells; it was increased 3. 337 times in hESCs derived RPE cells compared to hESCs derived cells containing pigmented foci; it increased 13. 574 times in hfRPE cells compared to hESCs-derived RPE cells, miR 211 does not change during differentiation from hESC to RPE, but it increased 44. 333 times in hESC derived RPE cells compared to hfRPE cells, miR 211 was the biggest difference in the miRNA expression pattern. In four cell types of hESCs, hESCs derived cells containing pigmented loci, hESCs- derived RPE cells and hfRPE cells, RT PCR showed the levels of miR 204 were 91.81±4.43, 2263.09± 206.39, 5996.80± 235.42, and 171676.45 ± 999.82 respectively, miR 204 was significantly increased during the whole course （t=18.22, 20.66, 279.38; F=0.001）. The levels ofmiR 211 were 2.23±0.31, 129.33±3.75, 125. 7592±4.78, and 16682.00±352.97 respectively, miR-211 was significantly increased from hESCs to cells containing pigmented foci and from hESCs derived RPE cells to hfRPE （t= 58.58, 81.24; P〈0.001）. Conclusion There is a continuous change of miR 204 and 211 in differentiation of RPE cells from hESCs.

关 键 词：胚胎干细胞/生理学 细胞分化/生理学 微RNAs/生理学 

分 类 号：R77[医药卫生—眼科]