Deciphering the mechanisms of tumorigenesis in human pancreatic ductal epithelial cells.
Deciphering the mechanisms of tumorigenesis in human pancreatic ductal epithelial cells.
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DOI：
10.1158/1078-0432.ccr-12-0032
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发表时间：
2013-02-01
期刊：
影响因子：
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通讯作者：
Chiao PJ
中科院分区：
文献类型：
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作者：
Chang Z;Li Z;Wang X;Kang Y;Yuan Y;Niu J;Wang H;Chatterjee D;Fleming JB;Li M;Abbruzzese JL;Chiao PJ
胰腺导管腺癌（PDAC）中最常见的遗传性病变已经确定。然而，我们对此类遗传改变如何协同作用诱导PDAC发育的理解仍然存在重大差距。在这项研究中，我们研究了在永生化的人胰腺导管上皮（HPDE）细胞系中的致瘤性转化的机制，依次引入PDAC签名改变到这个细胞系。突变K-ras，Her 2，p16/p14 shRNA，和Smad 4shRNA在HPDE细胞中的稳定表达的表型通过细胞增殖，迁移，侵袭，软琼脂和原位肿瘤发生的测定来检查。通过基因表达谱分析和途径分析进一步探讨其致瘤转化机制。转化细胞表现出增强的增殖，迁移和侵袭，显示锚定非依赖性生长在软琼脂中，原位生长肿瘤的PDAC的一些组织病理学特征。我们发现Smad 4在HPDE细胞的致瘤性转化中起关键作用。我们进一步发现MDM 2和Bmi-1在致瘤HPDE细胞中过表达，并且Bmi-1过表达受Smad 4调节。Ingenuity Pathway Analysis软件对微阵列数据的分析显示，整合素相关激酶（ILK）信号传导和细胞周期的失调是参与致瘤转化的最显着变化。总之，该细胞培养模型从基因损伤、特定信号通路的激活和一些组织病理学特征方面密切地概括了人类胰腺癌的发生。激活的K-ras和Her 2与失活的p16/p14和Smad 4的结合是转化HPDE细胞的充分和必要的，从而揭示了潜在的致瘤机制。
The most common genetic lesions in pancreatic ductal adenocarcinoma (PDAC) have been identified. However, significant gaps still exist in our understanding of how such genetic alterations act in concert to induce PDAC development. In this study, we investigated the mechanism of tumorigenic transformation in the immortalized human pancreatic ductal epithelial (HPDE) cell line by sequentially introducing PDAC signature alterations into this cell line. The phenotype for stable expression of mutant K-ras, Her2, p16/p14shRNA, and Smad4shRNA in HPDE cells was examined by assays for cell proliferation, migration, invasion; soft agar; and orthotopic tumorigenesis. The mechanisms of tumorigenic transformation were further explored by gene expression profiling and pathway analyses. The transformed cells exhibited enhanced proliferation, migration, and invasion; displayed anchorage-independent growth in soft agar; and grew orthotopic tumors with some histopathological features of PDAC. We found that Smad4 played key roles in the tumorigenic transformation of HPDE cells. We further found that MDM2 and Bmi-1 were overexpressed in the tumorigenic HPDE cells and that Bmi-1 overexpression was regulated by Smad4. Ingenuity Pathway Analysis software analysis of microarray data revealed that dysregulation of integrin-linked kinase (ILK) signaling and the cell cycle were the most significant changes involved in tumorigenic transformation. Altogether, this cell culture model closely recapitulated human pancreatic carcinogenesis from gene lesions, activation of specific signaling pathways, and some histopathological features. The combination of activated K-ras and Her2 with inactivated p16/p14 and Smad4 was sufficient and essential to transform HPDE cells, thus revealing the potential tumorigenic mechanism.