Rao M S, Ide H, Yeldandi A V, Kumar S, Reddy J K
Department of Pathology, Northwestern University Medical School, Chicago, IL 60611.
Carcinogenesis. 1994 Nov;15(11):2619-22. doi: 10.1093/carcin/15.11.2619.
We have examined ciprofibrate and dehydroepiandrosterone (DHEA)-induced hepatic lesions for the peroxisomal beta-oxidation system enzyme peroxisomal enoyl-CoA hydratase/3-hydroxyacyl-CoA dehydrogenase (PBE) and its mRNA using SDS-polyacrylamide gel electrophoresis, antibodies and cDNA probe. All 12 neoplastic nodules and nine hepatocellular carcinomas (HCCs) that were analyzed for PBE mRNA by in situ hybridization showed an intense signal comparable to the adjacent non-neoplastic liver. SDS-polyacrylamide gel electrophoresis of postnuclear fractions of six HCC and adjacent liver tissue showed a marked increase in an 80 kDa polypeptide. Immunoblot and Northern blot analysis showed a marked increase in PBE enzyme and PBE mRNA respectively in HCC and adjacent non-neoplastic liver tissue. In control livers (animals not treated with peroxisome proliferators), the levels of PBE enzyme and mRNA were very low or undetectable. The results of this study clearly indicate that peroxisome proliferator (PP)-induced liver lesions express peroxisomal enzymes to the same extent as adjacent liver and that these enzymes are not useful markers for identification of PP-induced lesions.
我们使用SDS-聚丙烯酰胺凝胶电泳、抗体和cDNA探针，检测了环丙贝特和脱氢表雄酮（DHEA）诱导的肝脏损伤中过氧化物酶体β-氧化系统酶过氧化物酶体烯酰辅酶A水合酶/3-羟基酰基辅酶A脱氢酶（PBE）及其mRNA。通过原位杂交分析PBE mRNA的所有12个肿瘤结节和9个肝细胞癌（HCC）均显示出与相邻非肿瘤性肝脏相当的强烈信号。对6个HCC及其相邻肝脏组织的核后组分进行SDS-聚丙烯酰胺凝胶电泳，结果显示一条80 kDa多肽显著增加。免疫印迹和Northern印迹分析分别显示HCC及其相邻非肿瘤性肝脏组织中PBE酶和PBE mRNA显著增加。在对照肝脏（未用过氧化物酶体增殖剂处理的动物）中，PBE酶和mRNA水平非常低或无法检测到。本研究结果清楚地表明，过氧化物酶体增殖剂（PP）诱导的肝脏损伤与相邻肝脏一样表达过氧化物酶体酶，并且这些酶不是鉴定PP诱导损伤的有用标志物。
