论文部分内容阅读
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 investigate the relationship between the changes of gene expression profiles of hydatidiform mole and choriocarcinoma (choriocarcinoma) and trophoblastic hyperplasia. Methods Gene expression profiling was performed on trophoblastic cells from 2 complete moles, 2 normal villi and primary cultured trophoblastic villi of early pregnancy and JAR cell line of choriocarcinoma using 4096 gene expression profiling chips. The expression of some enzymes related to DNA synthesis in normal villi, hydatidiform mole and choriocarcinoma JAR and JEG3 cells was confirmed by immunohistochemistry, western blot and reverse transcription polymerase chain reaction (RTPCR). Results In the two cases of hydatidiform mole specimens, 89 genes were significantly differentially expressed, accounting for 2.2% of the total number of genes. Compared with normal villi, hydatidiform mole significantly increased (up-regulated) 24 genes and significantly decreased (down-regulated) 65 genes. Compared with normal primary cultured trophoblast cells, 432 genes were down-regulated and 380 genes were up-regulated in JAR cells. Mole and choriocarcinoma cells, a total of 46 down-regulated genes, a common gene up 13. In hydatidiform mole and choriocarcinoma, genes that inhibit cell growth are downregulated, whereas genes associated with cell proliferation, malignant transformation, metastasis and resistance are up-regulated. Immunohistochemistry, Western blotting and RTPCR demonstrated that expression of thymidine kinase 1 and nucleotide reductase small subunits associated with DNA synthesis were significantly increased in hydatidiform mole, choriocarcinoma JAR and JEG3 cells. Conclusion There are abnormally expressed genes in hydatidiform mole and choriocarcinoma. Trophoblast hyperplasia may be related to the abnormally high expression of DNA synthetase.