印迹基因 H19对人绒毛膜上皮癌细胞JEG-3基因表达谱的影响

摘要

Objective To obtain the expression pattern of imprint gene H19 in JEG-3 cell in order to explore the regulation mechanism of H19 on trophoblast cellular biological behavior .Methods After correct identification with sequencing for the recom-binant eukaryotic expression plasmid pRc/CMV which including the whole length of H19 cDNA ,the plasmid was transfected to the cell line JEG-3 .The expression of H19 mRNA was observed and the gene expression profile of three groups of JEG-3 cell were de-tected with Affymetri :U133 plus 2 .0 Array .Results After being transfected with target H 19 gene ,the expression of the mRNA level was significantly increased compared with control group .And the gene expression profile was changed significantly .19 genes were up-regulated ,77 genes were down-regulated .Expression levels of HES1 gene which being choosed as a different expression gene were detected by fluorescence quantitative PCR in severe preeclampsia placenta tissue and normal late pregnant placenta .The expression level of HES1 mRNA in severe preeclampsia placenta decreased significantly than normal late pregnant placenta tissues . Conclusion Many genes induced by H19 have been screened by high-throughput gene chip method .It provides the experimental ba-sis for advanced studying the regulation the cellular biological behavior with H 19 gene .%目的：获得印迹基因H19对滋养细胞株人绒毛膜上皮癌细胞系（JEG-3）细胞基因表达谱的影响，以期探讨 H19对滋养细胞生物学行为的调控机制。方法含人全长H19 cDNA 重组真核表达质粒pRc/CMV 经测序鉴定正确后，转染JEG-3细胞，采用Affymetrix的U133 plus 2．0 Array基因表达谱芯片检测基因表达谱的变化。结果转染后H19 mRNA表达显著升高， JEG-3细胞基因表达谱明显改变，共筛选出96条差异基因，其中上调基因19条，下调基因77条。选择其中一种差异表达基因HES1，采用荧光定量PCR检测发现重度子痫前期胎盘组织HES1 mRNA的表达水平较正常晚孕胎盘组织中HES1 mRNA的表达水平明显降低。结论高通量基因芯片筛选出了H19调控滋养细胞的相关基因，为进一步探讨 H19对滋养细胞生物学行为的调控机制提供了实验基础。