Molecular biological analysis in the process of carcinogenesis of hydatidiform mole cells transformed by tumor promoter agents

本研究所用的磨牙细胞系(BM-72)是从一例完全性葡萄胎患者的磨牙囊中培养出来的。对致癌促进剂(TPA)诱导葡萄胎细胞恶性转化的能力进行了检测，包括在双层软琼脂上的培养效率和致瘤性。在检测TPA的作用时，细胞在含有TPA(0.1，1.0，10,100pg/ml)的培养液中培养2周。在将TPA处理的细胞移植到SCID小鼠体内2周后，观察到了肿瘤的诱导，并且在2周的治疗组中发现了肿瘤形成的增加，其反应与浓度相关。RAP-PCR差异显示技术可用于获得基因表达随不同条件变化的基因。我们用mRNA差异显示的方法鉴定了致癌促进剂TPA(0.1，1.0，10,100pg/ml)处理BM-72细胞2周后其表达发生变化的基因。在本实验中，与对照组和其他浓度的TPA相比，在含100 pg/mlTPA的培养液中培养的磨牙细胞的cDNA中有两个克隆没有表达。

The cultured cell line of molar cells employed in this study (BM-72) was driven from molar cysts obtained from a patient with complete hydatidiform mole. The carcinogenic promoter (TPA) was tested for the abillity to induce malignant transformation in cultured hydatidiform mole cells in terms of plating efficiency in double-layr soft agar and tumorigenesis. In testing for the effects of TPA,the cells were cultured in medium containing TPA (0.1,1.0,10,100 pg/ml) for 2 weeks. The tumorigenetic induction was seen following transplantation of 2 weeks TPA-treated cells into SCID mice and increase in tumorigenesis was noted in the 2-week treatment group with a response correlated to concentration. The technique of RAP-PCR differential display is useful for obtaining genes whose expression changes with various conditions. we have used the mRNA differential display method to identify genes whose expression is altered as a consequence of the transformation of BM-72 cultued cells treated by carcinogenic promoter, TPA (0.1,1.0,10,100 pg/ml), for 2 weeks. In this display, two clones were found to be not expressed in the cDNA of molar cells cultured in medium containing with 100 pg/ml of concentration of TPA in comparison to control and other concentration of TPA.