Modification of DNA damage detecting method, Terminal transferase dependent PCR(TDPCR), and its application to endometrial carcinogenesis

雌激素在子宫内膜癌发生中的作用是有争议的，因此，我们拟用末端转移酶依赖的、方法检测雌激素或其代谢产物对子宫内膜细胞的损伤作用。首先，我们应用毛细管测序仪对TDPCR进行了改进，从而避免了复杂、耗时的过程。通过改良的TDPCR，我们发现邻苯二酚类雌激素对子宫内膜癌中频繁突变的基因PTEN有DNA损伤作用，而雌激素不能。此外，DNA损伤部位与子宫内膜癌中已知的PTEN突变部位相对应。综上所述，这些数据表明，雌激素的代谢产物在子宫内膜癌的发生中与PTEN突变有关。

The involvement of estrogen in endometrial carcinogenesis is controversial、therefore, we intended to examine estrogen or its metabolite could induce DNA damage on endometrial cells by using terminal transferase dependent PCR(TDPCR) method. At first, we modified TDPCR by applying a capillary sequencer and consequently we could avoid its complicated, time-consuming process. By using modified TDPCR, we found that catechol estrogen caused DNA damages on frequently mutated gene in endometrial carcinoma, PTEN, while estrogen did not. In addition, the foci of DNA damage corresponded to known mutation site of PTEN in endometrial carcinoma. Taken together, these data suggest that metabolite of estrogen is responsible for PTEN mutations in endometrial carcinogenesis.