Tsujiuchi Toshifumi, Sasaki Yasutaka, Konishi Yoichi, Tsutsumi Masahiro
Department of Oncological Pathology, Cancer Center, Nara Medical University, Kashihara, Nara, Japan.
Mol Carcinog. 2002 Oct;35(2):57-62. doi: 10.1002/mc.10074.
Alteration of the retinoblastoma-related gene RB2/p130 was investigated in lung adenocarcinomas induced by N-nitrosobis(2-hydroxypropyl)amine (BHP) in male Wistar rats. At 6 wk of age, 21 animals were given 2000 ppm of BHP in their drinking water for 12 wk and then maintained without further treatment until they were killed at the end of week 25. A total of 21 lung adenocarcinomas were obtained, and total RNAs were extracted from each for mutation analysis of RB2/p130 by the reverse transcription-polymerase chain reaction-single-strand comformation polymorphism approach. No mutations were found in exons 19-22. However, examination of the expression of the RB2/p130 gene by Northern blot analysis showed mRNA levels to be significantly lower than those of normal lung tissues. Western blot analysis showed reduction of the pRb2/p130 protein in all of the adenocarcinomas examined. These results suggest that alteration of the RB2/p130 gene may play important roles in the development of lung adenocarcinomas induced by BHP in rats.
在雄性Wistar大鼠中，研究了由N-亚硝基双（2-羟丙基）胺（BHP）诱导的肺腺癌中视网膜母细胞瘤相关基因RB2/p130的改变。6周龄时，给21只动物饮用含2000 ppm BHP的水12周，然后不再进行进一步处理，直至在第25周结束时处死。共获得21个肺腺癌，从每个肿瘤中提取总RNA，通过逆转录-聚合酶链反应-单链构象多态性方法对RB2/p130进行突变分析。在第19-22外显子中未发现突变。然而，通过Northern印迹分析检测RB2/p130基因的表达，结果显示mRNA水平显著低于正常肺组织。蛋白质印迹分析显示，在所检测的所有腺癌中，pRb2/p130蛋白均减少。这些结果表明，RB2/p130基因的改变可能在BHP诱导的大鼠肺腺癌发生中起重要作用。
