Objective To identify specific expressed mRNA of rat hepatic stem cells to discover the markers using gene chip technology and confirm them using immunohistochemistry. Methods Two Pregnant rats at embryonic day(ED) 14 and two adult rats were collected as specimens. RNA was extracted and cDNA was synthesized by reverse transcription, then cRNA was synthesized by transcription in vitro and making mark at the same time. cRNA was hybridizated, washed, dyed after purification and chip was scanned. Data of gene ship were processed and analyzed, and to find the objective gene. Immunohistochemistry staining was used to detect the expressions of HMGA1 protein in embryonic liver tissues. Results According to screening standard, 787 genes which were overexpressed in rat embryonic liver. These 787 genes were divided into eight groups. Immunohistochemistry staining showed that HMGA1 was positive in most embryonic liver cells at the ED 14 and mainly distributed in the cytoplasm. Conclusions Differential expression genes in embryonic liver tissues are scanned by gene chip. Immunohistochemistry staining shows that the high mobility group protein Al is the specific marker of rat liver embryonic stem cells.%目的 筛选大鼠胚胎肝脏组织特异性表达的mRNA,寻找肝干细胞的特异性标记物.方法 取成年大鼠肝脏2个、2周胚龄大鼠胚胎肝脏2个作为标本.提取标本的RNA,反转录合成cDNA,再体外转录合成cRNA,同时进行标记.将cRNA纯化后进行杂交、洗涤、染色,对芯片进行扫描.将基因芯片数据进行处理与分析,筛选出目的基因.免疫组化方法检测各相关蛋白在胚胎肝脏组织中的表达.结果 筛选出787个在大鼠胚胎肝脏组织中高表达的差异表达基因,涉及8个类别.免疫组化检测发现,高迁移率族蛋白A1(HMGA1)在2周胚龄多数胚肝胎细胞中表达阳性,阳性信号多分布于细胞质.结论 大鼠胚胎肝脏组织发育相关的差异表达基因筛选可采用基因芯片,HMGA1是大鼠胚胎肝干细胞特异性标记物.
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