Role of Oncogene and Cytochrome P450 in Carcinogenesis of Gynecologic carcinoma

点击翻译按钮获取中文摘要

Most carcinogens are bioactivated by cytochrome P450(CYPs) and these enzymes within target cells are closely related to susceptibility to cancer. Since extrahepatic CYPs are typically at much lower levels, the existence and the role of CYP in extrahepatic tissues have been difficult to assess. In this study, we modified the reverse transcriprtase-ploymerase chain reaction (RT-PCR) to evaluate the quantities of CYP 2E1 mRNA in human endometrium. Total RNA from human endometrium was reverse transcribed and coamplified by PCR in the same tube containing both primer pairs of CYP 2E1 and beta-actin. The CYP 2E1 and beta-actin PCR products were 298 bp and 600 bp, respectively. The restriction enzyme mboi digested these two products to the predicted size for DNA fragments, demonstrating that both PCR products were specific and CYP 2E1 mRNA exists in human endometrium. CYP 1A1 mRNA was also examined, m but could not be detected clearly. Adding [alpha-^<32>P]dCTP to reaction mixture made it possible to quantify the relative yield of the CYP 2E1 PCR product in comparison with the beta-actin product. The ratio of the yield of the CYP 2E1 PCR product comparison with the beta-actin PCR product could be calculated at a point of 25 cycles of amplification. This ratio and serum estradiol levels were correlated positively (gamma=0.654, P<0.05), but not relationship to serum progesterone Levels was observed.