Abstract: 【Abstract】 Objective To investigate the effect of Bmi-1 gene overexpression on the proliferation of human pancreatic ductal epithelial cell line hTERT-HPNE. Methods〓The plasmid containing Bmi-1 gene and control was transducted into hTERT-HPNE cells by retroviral transfect. The expression of Bmi-1 mRNA and protein levels were detected by quantitative real-time PCR (qRT-PCR) analysis and Western blot analysis, respectively. The effect of Bmi-1 gene overexpression on the proliferation, migration, invasion and colony formation by MTS assay, Transwell assay and soft agar assay. Results The results of qRT-PCR analysis and Western blot analysis demonstrated that stable transfected cell lines successfully established..The expression of Bmi-1 promoted cell growth and proliferation, enhanced clone formation capability, and increased migration and invasion abilities. Conclusion〓Our findings demonstrated that ectopic expression of Bmi-1 lead to malignant transformation of pancreatic ductal epithelial cells.

Key words: Pancreatic ductal epithelial cell, Malignant transformation, Bmi-1

摘要： 【摘要】 目的 探讨原癌基因B细胞特异性莫洛氏鼠白血病病毒插入位点-1(Bmi-1)过表达对人正常胰腺上皮细胞株hTERT-HPNE恶性转化的影响。方法〓采用逆转录病毒介导转染方法将携带原癌基因Bmi-1的质粒或空质粒稳定转染hTERT-HPNE,通过Real-time PCR及Western blot在mRNA及蛋白水平鉴定转染效果。采用MTS法、Traswell小室法、软琼脂克隆形成试验检测稳定转染Bmi-1对hTERT-HPNE细胞增殖、迁移、侵袭和软琼脂克隆形成能力的影响。结果〓Real-time PCR及Western blot结果均表明成功建立稳定转染Bmi-1基因的hTERT-HPNE细胞株。过表达Bmi-1基因使hTERT-HPNE细胞增殖、迁移、侵袭和软琼脂克隆形成能力明显增高。结论〓在细胞水平过表达Bmi-1基因恶性转化hTERT-HPNE细胞。

关键词: 胰腺导管上皮细胞, Bmi-1, 恶性转化