Involvement of PSMD10, CDK4, and Tumor Suppressors in Development of Intrahepatic Cholangiocarcinoma of Syrian Golden Hamsters Induced by Clonorchis sinensis and N-Nitrosodimethylamine.
Involvement of PSMD10, CDK4, and Tumor Suppressors in Development of Intrahepatic Cholangiocarcinoma of Syrian Golden Hamsters Induced by Clonorchis sinensis and N-Nitrosodimethylamine.
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DOI：
10.1371/journal.pntd.0004008
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发表时间：
2015
影响因子：
3.8
通讯作者：
Hong ST
中科院分区：
文献类型：
--
作者：
Uddin MH;Choi MH;Kim WH;Jang JJ;Hong ST
华支睾吸虫是胆管癌（CCA）的I组生物致癌物。尽管流行病学证据将华支睾吸虫病和CCA联系起来，但对这一过程中涉及的潜在分子机制知之甚少。在本研究中，我们研究了癌基因和肿瘤抑制因子，包括PSMD 10，CDK 4，p53和RB在C.中华仓鼠CCA模型。采用不同的组织化学/免疫组织化学技术检测4组仓鼠的CCA：未感染对照组（Ctrl.），感染C.中华绒螯蟹（Cs）、摄入N-亚硝基二甲胺（NDMA）和既感染Cs又引入NDMA（Cs + NDMA）。通过定量PCR（qPCR）和蛋白质印迹法分析来自所有组的肝组织的基因/蛋白质表达。Cs + NDMA组所有仓鼠均可见CCA，高、中、低分化型分别占组织切片总面积的21.8% ± 1.5%、13.3% ± 1.3%和10.8% ± 1.3%。从感染第8周开始，所有CCA分化均以时间依赖性方式进行。CCA间质的特征是I型胶原、粘蛋白和增殖细胞核抗原（PCNA）增加。qPCR分析显示Cs + NDMA组中PSMD 10、CDK 4和p16 INK 4过表达，而p53低表达。我们没有观察到RB1在mRNA水平上的变化，但发现RB蛋白的显著下调。凋亡相关基因BAX和caspase 9在CCA组织中表达下调。除NDMA组外，其余各组的基因/蛋白表达与病理变化相吻合。虽然NDMA组的仓鼠没有表现出明显的病理学病变，但我们观察到Akt/PKB和p53基因的过度表达，表明该组中的分子相互作用可能与该动物模型中CCA的启动有关。提示癌基因PSMD 10和CDK 4及抑癌基因p53和RB参与了C.中华绒螯蟹诱导仓鼠CCA。 华支睾吸虫是一种蠕虫寄生虫，是人类胆管癌（CCA）或胆管癌的致癌因子。尽管大量令人信服的证据表明C.中华绒螯蟹和CCA的遗传/蛋白质组水平的机制知之甚少。为了探索潜在的分子机制，我们研究了C.中华仓鼠CCA模型。这里C.用N-亚硝基二甲胺诱导中华绒螯蟹成功地诱导了所有仓鼠的CCA。组织病理学检查证实了CCA的发生，并检测到过量的胶原纤维、粘蛋白和细胞分裂相关蛋白。定量PCR分析显示癌基因PSMD 10、CDK 4水平升高，抑癌基因p53水平降低。蛋白质印迹分析观察到另一种称为RB蛋白的肿瘤抑制因子显著减少。基因/蛋白表达与CCA仓鼠的病理变化相吻合。本研究提示，癌基因PSMD 10和CDK 4、抑癌基因p53和RB蛋白参与了C.中华绒螯蟹诱导仓鼠CCA。
Clonorchis sinensis is a group-I bio-carcinogen for cholangiocarcinoma (CCA). Although the epidemiological evidence links clonorchiasis and CCA, the underlying molecular mechanism involved in this process is poorly understood. In the present study, we investigated expression of oncogenes and tumor suppressors, including PSMD10, CDK4, p53 and RB in C. sinensis induced hamster CCA model. Different histochemical/immunohistochemical techniques were performed to detect CCA in 4 groups of hamsters: uninfected control (Ctrl.), infected with C. sinensis (Cs), ingested N-nitrosodimethylamine (NDMA), and both Cs infected and NDMA introduced (Cs+NDMA). The liver tissues from all groups were analyzed for gene/protein expressions by quantitative PCR (qPCR) and western blotting. CCA was observed in all hamsters of Cs+NDMA group with well, moderate, and poorly differentiated types measured in 21.8% ± 1.5%, 13.3% ± 1.3%, and 10.8% ± 1.3% of total tissue section areas respectively. All CCA differentiations progressed in a time dependent manner, starting from the 8th week of infection. CCA stroma was characterized with increased collagen type I, mucin, and proliferative cell nuclear antigen (PCNA). The qPCR analysis showed PSMD10, CDK4 and p16INK4 were over-expressed, whereas p53 was under-expressed in the Cs+NDMA group. We observed no change in RB1 at mRNA level but found significant down-regulation of RB protein. The apoptosis related genes, BAX and caspase 9 were found downregulated in the CCA tissue. Gene/protein expressions were matched well with the pathological changes of different groups except the NDMA group. Though the hamsters in the NDMA group showed no marked pathological lesions, we observed over-expression of Akt/PKB and p53 genes proposing molecular interplay in this group which might be related to the CCA initiation in this animal model. The present findings suggest that oncogenes, PSMD10 and CDK4, and tumor suppressors, p53 and RB, are involved in the carcinogenesis process of C. sinensis induced CCA in hamsters. Clonorchis sinensis is a helminth parasite and a carcinogenic agent for cholangiocarcinoma (CCA) or bile duct cancer in humans. Though a large and compelling body of evidence suggests an association between C. sinensis and CCA, the mechanism underlying at the genetic/proteomic level is little known. To explore the underlying molecular mechanism we investigated a number of genes/proteins in C. sinensis induced hamster CCA model. Here C. sinensis induced CCA successfully in all hamsters when introduced with N-nitrosodimethylamine. The histopathology confirmed the development of CCA and detected excessive collagen fibers, mucin and cell division related protein. The quantitative PCR analysis showed increased levels of oncogenes PSMD10, CDK4 and decreased level of tumor suppressor gene p53. The western blot analysis observed significant decrease of another tumor suppressor called RB protein. Genes/protein expressions were matched well with the pathological changes of CCA hamster. The present study suggests that oncogenes, PSMD10 and CDK4, and tumor suppressors gene p53 and protein RB, are involved in the carcinogenesis process of C. sinensis induced CCA in hamsters.