Role of Oncogene and Cytochrome P450 in Carcinogenesis of Gynecologic carcinoma

大多数致癌物被细胞色素P450（CYP）生物激活，靶细胞内的这些酶与癌症的易感性密切相关。由于肝外CYP通常处于低得多的水平，因此难以评估肝外组织中CYP的存在和作用。本研究应用逆转录-聚合酶链反应（RT-PCR）技术检测人子宫内膜组织中E_2E_1 mRNA的表达。从人子宫内膜的总RNA逆转录和共扩增PCR在同一管含有两个引物对的β-肌动蛋白和β-肌动蛋白。PCR扩增产物分别为298 bp和600 bp。限制性内切酶mboi将这两种产物消化至DNA片段的预期大小，证明两种PCR产物都是特异性的，并且人子宫内膜中存在E1 mRNA。还检测了β 1 A1 mRNA，但不能清楚地检测到。向反应混合物中加入[α-β-<32>P]dCTP使得有可能定量与β-肌动蛋白产物相比的β 2 E1 PCR产物的相对产率。可以在25个扩增循环的点处计算与β-肌动蛋白PCR产物相比的β-肌动蛋白2 E1 PCR产物的产率比率。该比值与血清雌二醇水平呈正相关（γ =0.654，P&lt;0.05），与血清孕酮水平无相关性。

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.