Department of Gastroenterology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, 197 Ruijin Er Road, Shanghai 200025, China.
Anticancer Agents Med Chem. 2013 Feb;13(2):261-9. doi: 10.2174/1871520611313020012.
Pancreatic ductal adenocarcinoma (PDAC) is an almost lethal disease. Thus, it is important to better understand its genetic progression from normal cells through precancerous lesions pancreatic intraepithelial neoplasia (PanIN) to invasive pancreatic cancer. Carriers of a germline mutation in BRCA2 have an increased risk of developing PDAC when compared with the general population. The purpose of our study was to examine the role of BRCA2 dysfuction in the progression of PDAC. Here we generated a novel in vitro model of pancreatic carcinogenesis. Cancerous PanIN-BR1 cells were established by stable transduction with lentiviral-mediated BRCA2 RNA interference in PanIN cell isolated from mice with oncogenic KrasG12D. Our data showed that silencing of BRCA2 promoted cell proliferation, migration and invasion in vitro. The tumorigenic potential of PanIN-BR1 were assessed by xenograft tumor formation in BALB/c nude mice. The expression of PCNA , Snail and Slug in the tumor xenografts was detected by immunohistochemistry. The staining for PCNA, Snail and Slug in PanIN-BR1-formed xenograft tissue was significantly more intense than PanIN-formed xenograft tissue. Microarray assay was also performed. Based on gene expression profiling and further validation by real-time PCR and Western Blot, we found that the expression of Cyclinb2, Cyclina2, Twist1, Wisp1 and Cxcr4 revealed a significant increase in the PanIN-BR1 cells, however, the expression of p15, p16 and Wisp2 showed a significant decrease in the PanIN-BR1 cells, compared to the PanIN cells. Collectively, these data reported here demonstrate that BRCA2 may be a promising therapeutic targets for PDAC progression.
胰腺导管腺癌（PDAC）是一种几乎致命的疾病。因此，更好地了解其从正常细胞经过癌前病变胰腺上皮内瘤变（PanIN）到侵袭性胰腺癌的遗传进展非常重要。与普通人群相比，BRCA2 种系突变的携带者患 PDAC 的风险增加。我们研究的目的是研究 BRCA2 功能障碍在 PDAC 进展中的作用。在这里，我们建立了一种新的胰腺癌变体外模型。通过携带致癌性 KrasG12D 的小鼠中分离的 PanIN 细胞中，通过慢病毒介导的 BRCA2 RNA 干扰稳定转导，建立了癌症 PanIN-BR1 细胞。我们的数据表明，BRCA2 沉默在体外促进了细胞增殖、迁移和侵袭。通过在 BALB/c 裸鼠中进行异种移植肿瘤形成来评估 PanIN-BR1 的致瘤潜力。通过免疫组织化学检测肿瘤异种移植组织中 PCNA、Snail 和 Slug 的表达。PanIN-BR1 形成的异种移植组织中 PCNA、Snail 和 Slug 的染色明显强于 PanIN 形成的异种移植组织。还进行了微阵列分析。基于基因表达谱，通过实时 PCR 和 Western Blot 进一步验证，我们发现 Cyclinb2、Cyclina2、Twist1、Wisp1 和 Cxcr4 的表达在 PanIN-BR1 细胞中明显增加，然而，与 PanIN 细胞相比，p15、p16 和 Wisp2 的表达在 PanIN-BR1 细胞中明显降低。总之，这些数据表明 BRCA2 可能是 PDAC 进展的有前途的治疗靶点。
