葡萄胎和绒毛膜癌基因表达谱改变与滋养细胞增生的关系

Changes in gene expression profiles of hydatidiform mole and choriocarcinoma as compared with trophoblast hyperplasia

目的研究葡萄胎和绒毛膜癌(绒癌)基因表达谱改变与滋养细胞增生的关系。方法用含4096条基因的表达谱芯片,对2例完全性葡萄胎、2例正常绒毛及原代培养的早孕绒毛滋养细胞和绒癌JAR细胞株滋养细胞进行基因差异表达谱分析。用免疫组化、免疫印迹和反转录聚合酶链反应(RTPCR)方法,对正常绒毛和葡萄胎组织及绒癌JAR和JEG3细胞中与DNA合成有关的一些酶表达水平进行印证。结果2例葡萄胎标本中,均有显著差异表达基因89条,占基因总数的2.2%。葡萄胎与正常绒毛相比,24条基因表达显著增高(上调),65条基因显著降低(下调)。JAR细胞与正常原代培养滋养细胞相比,432条基因下调,380条基因上调。葡萄胎和绒癌细胞中,共同下调基因有46条,共同上调基因有13条。在葡萄胎和绒癌中,抑制细胞生长的基因下调,而与细胞增生、恶性转化、转移及耐药等有关的基因上调。经免疫组化、免疫印迹和RTPCR方法证实,在葡萄胎、绒癌JAR和JEG3细胞中,与DNA合成有关的胸腺嘧啶核苷激酶1和核苷酸还原酶小亚基的表达显著增高。结论葡萄胎和绒癌存在着异常表达基因,滋养细胞增生可能与DNA合成酶异常高表达有关。

Objective To study the relationship of changes in gene expression profiles of hydatidiform mole and choriocarcinoma with hyperplasia of trophoblasts. Methods The differentially expressed genes were analyzed in two pairs of tissues of hydatidiform mole versus normal villi, and in two pairs of normal primary culture trophoblasts versus JAR cell line of chariocarcinoma, using cDNA microarray containing 4096 genes. To confirm the results of cDNA microarray analysis, expressions of some up-regulated genes related to DNA synthesis in ~normal villi, hydatidiform mole, and 2 choriocarcinoma cell lines (JAR and JEG-3) were examined by immunohistochemistry, immunoblotting and RT-PCR. Results A total of 89 genes were differentially expressed in all hydatidiform moles, accounting for 2.2% of the genes arrayed. Of the 89 genes, 24 were up-regulated and 65 were down-regulated. Compared with normal primary trophoblasts, there were 433 genes up-regulated and 380 genes down-regulated in JAR cell line. Forty six genes were up-regulated in both hydatidiform mole and choriocarcinoma,while 13 genes were down-regulated. Some genes associated with cell proliferative inhibition were significantly down-regulated, whereas those associated with cell proliferation, malignant transformation, metastasis and drug resistance were highly up-regulated. The expressions of thymidine kinase 1, the small subunit of ribonucleotide reductase (RRM2) were significantly increased in hydatidiform mole, JAR and JEG-3 cells. Conclusion Abnormal expression of genes exists in hydatidiform mole and choriocarcinoma. Hyperplasia of trophoblasts may be related to over-expression of genes coding for synthetic enzymes.