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.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
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)
关键词
癌
肝细胞
NDRG2
转染
Carcinoma, hepatocellular
NDRG2
Transfeetion