稳定转染NDRG2基因对人肝癌细胞HepG2的作用  

Inhibitory effects of NDRG2 gene on HepG2 cells after stable gene transfection

作　　者：吴国强[1] 李开宗[2] 窦科峰[2] 药立波[3] 刘新平[3] 

机构地区：[1]沈阳军区总医院普通外科,110016 [2]第四军医大学西京医院肝胆外科 [3]第四军医大学基础部生化与分子生物学教研室

出　　处：《中华实验外科杂志》2011年第12期2127-2130,共4页Chinese Journal of Experimental Surgery

基　　金：基金项目：国家863计划资助项目（2006AA022194）

摘　　要：目的观察NDRG2基因（N—myc downstream regulated gene2）对肝癌细胞株HepG2生物学行为的影响。方法采用脂质体法将正义及反义NDRG2基因稳定转染HepG2及QZG细胞，采用逆转录．聚合酶链反应（RT-PCR）、免疫组织化学及Western blot法进行鉴定；比较观察其生物学特性的变化。结果NDRG2基因在NDRG2-HepG2细胞中得到了稳定表达；NDRG2-HepG2与HepG2细胞在细胞计数（1、2、3、4、5、6、7d分别为3×10^5／6×10^5、4×10^5／12×10^5、5×10^5／13×10^5、6．0×10^5／13．5×10^5、6．5×10^5／15．0×10^5、6．8×10^5／16．5×10^5、7×10^5／17×10^5）、细胞集落形成（32．4±4．7／56．5±5．6）及成瘤重量（0．0462g／0．2120g）差异有统计学意义（P〈0．05）；而Anti—QZG与QZG细胞的差异无统计学意义（P〉0．05）。结论NDRG2基因可抑制肝癌细胞株HepG2细胞生长增殖。Objective To investigate the biological effects of the NDRG2 gene on HepG2 cells. Methods The sense and anti-sense NDRG2 full length cDNA were transfected into HepG2 and QZG cell lines by the vectors of pIRES2-EGFP-NDRG2 and pIRES2-EGFP-Anti-NDRG2 respectively and the transfec- tants were obtained by clone selection. The expression levels of NDRG2 gene were detected by reverse transcription-polymerase chain reaction （RT-PCR）, immunochemistry and Western blotting. Effects of NDRG2 and Anti-NDRG2 transfection on the biological characteristics of HepG2 and QZG cells were analyzed, including proliferation rate, colony-forming efficiency （CFE） and tumor formation. Results The NDRG2-HepG2 cell line was obtained after stable gene transfection. There was significant difference between NDRG2-HepG2 and HepG2 cells （P 〈 0. 05） in the cell count （ 1, 2, 3, 4, 5, 6, 7 days after passage culture ： 3 × 10^5 vs. 6×10^5,4×10^5 vs. 12×10^5, 5×10^5 vs. 13×10^5, 6.0×10^5 vs. 13.5×10^5, 6.5×10^5 vs. 15.0×10^5, 6. 8 ×10^5 vs. 16. 5 ×10^5, 7 ×10^5 vs. 17×10^5 respectively）. There was no significant difference in the CFE （32. 4 ±4. 7 vs. 56. 5 ±5.6） and the tumor formation （0. 0462 g vs. 0. 2120 g） between Anti-QZG and QZG cells （P 〉 0. 05）. Conclusion NDRG2 gene can arrest the growth of HepG2 cells.

关 键 词：癌 肝细胞 NDRG2 转染 

分 类 号：R735.7[医药卫生—肿瘤]