腺病毒介导的p27mt基因转移对肝癌细胞生长的影响

目的 :研究突变型p2 7基因 (p2 7mt)对肝癌细胞生长的影响。方法 :采用重组腺病毒载体Ad p2 7mt将p2 7mt全长cDNA转入到肝癌细胞系SMMC 772 1中。3H TdR掺入法及软琼脂集落形成实验检测p2 7mt对肝癌细胞增殖的作用。结果 :转染p2 7mt基因的SMMC 772 1细胞在蛋白质水平有高水平的基因表达。其3H TdR掺入量及集落形成率明显低于对照组 (P <0 .0 5)。结论 :p2 7mt基因能抑制肝癌细胞增殖。p2 7mt可作为目的基因用于基因治疗。

Effects of Adenovirus Mediated Human p27mt on the Growth in Human Hepatocellular Carcinoma Cell Lines SMMC-7721

Objective:To study the effects of P27mt gene on growth of huma n hepatocellular carcinoma cells.Method:A replication deficient adenovirus vector encoding p27mt (Ad-p27mt) was used and the full length cDNA of p27mt was transfected into h uman SMMC-7721 cell lines in vitro.Effects of P27mt on proliferation was obser ved by 3H-thymidine incorporation and anchorage independent growth in sof t agar.Results:The virus titer is 7.95×10 12cfu/ml.The transd uction efficiency is nearly 100% when multiplicity of infection ≥50,The expre ssion of p27mt increased significantly in SMMC-7721 cells transfected with p27m t. 3H-thymidine incorporation demonstrated the synthesis of DNA in SMMC-7 721 cells is inhibited.The rates of anchorage independent growth in soft agar d ecreased significantly.Conclusion:p27mt gene could inhibit SMMC-7721 cells proliferat ion,p27mt gene can be used as an agent for gene therapy.