【摘要】 目的　探讨野生型 /突变型RASSF1A基因的表达对人肝癌细胞株QGY 770 3在体内外生长的影响。方法　构建突变型RASSF1A基因的真核表达载体 ,通过脂质体介导的基因转染法将野生型 /突变型RASSF1A基因的真核表达载体及空载体转染肝癌细胞株QGY 770 3 ,G418筛选稳定表达株 ,并以逆转录 聚合酶链反应 (RT PCR)和Westernblot进行鉴定。通过细胞生长曲线、软琼脂集落形成试验和裸鼠致瘤性试验对各稳定表达株的生物学行为进行检测。结果　成功建立稳定表达野生型和突变型RASSF1A基因的肝癌细胞株。以转染空载体的QGY 770 3细胞为对照 ,野生型RASSF1A基因的表达可明显抑制肝癌细胞在体外的生长 ,显著降低肝癌细胞在软琼脂中形成的克隆数目 (P <0 .0 1)和大小 ,显著减慢裸鼠皮下瘤的生长速度和重量 (P <0 .0 1)。突变型RASSF1A基因的表达对肝癌细胞的生长影响不大。结论　野生型RASSF1A的重表达有助于QGY 770 3恶性表型的逆转 ;野生型RASSF1A基因是一个肝癌相关的抑癌基因。更多还原

【Abstract】 Objective To explore the effect of wild type or mutant RASSF1A gene expression on the growth of human hepatocellular carcinoma (HCC) cell line QGY 7703.Methods The mutant RASSF1A gene eukaryotic expressing vector was constructed.The RASSF1A (wt or mutant) expression vector and empty vector was transferred into HCC cell line QGY 7703 with LipofectAMINE 2000 reagents.The positive clones that expressed RASSF1A gene stably were chosen by G418 and checked by reverse transcription polymerase chain reaction (RT PCR) and Western blot.The cytobiological behaviors of those positive clones was analyzed by using cell growth curve,anchorage independent growth assay and tumorigenesis in nude mice.Results The HCC cell lines that expressed wide type or mutant RASSF1A gene stably were established successfully.The growth of wide type RASSF1A expressed cells was obviously inhibited on dishes compared with the control cells transfected with empty vectors.The wide type RASSF1A expressed cells formed smaller and significantly fewer colonies than the control cells.The volume and the weight of tumor formed by wide type RASSF1A expressed cells in nude mice were also significantly reduced.Mutant RASSF1A gene expression had little effect on the growth of QGY 7703 cells in vitro and in vivo.Conclusion The re expression of wild type RASSF1A could favor the malignant phenotype revision of QGY 7703 cells and RASSF1A gene might be a good candidate of tumor suppressor gene related with HCC.更多还原