肝癌细胞内表达抗肝癌单链抗体对肝癌细胞的杀伤作用

为研究抗肝癌单链抗体（scFV）基因在肝癌细胞内表达对肝癌细胞的作用，采用脂质体转染技术把含抗人肝癌scFV基因的逆转录病毒载体导入人肝癌细胞系HCC9204中。G418筛选阳性细胞克隆。RNAdotblot杂交分析scFV基因在HCC细胞中的表达。光镜及电镜技术观察转染细胞结构的变化。流式细胞仪检测转染细胞的增殖活性。结果：抗人肝癌scFV在人肝癌细胞系HCC9204中稳定有效表达。流式细胞测定出现亚G1期死亡细胞峰；光镜下表现为细胞疏松，胞浆空泡变性，瘤巨细胞增多。电镜分析表现为细胞表面微绒毛显著减少，线粒体及内质网空泡变性，细胞死亡明显。提示：肝癌细胞内表达抗肝癌scFV对肝癌细胞有明显杀伤作用，为肝癌基因治疗提供一种新方法。

INTRACELLULAR EXPRESSION OF ANTI HEPATOCELLULAR CARCINOMA (HCC) SINGLE CHAIN VARIABLE REGION FRAGMENT (sCFv) IN HCC CELLS INDUCES CELL DEATH

Objective; Study the effects of intracellular expression of anti HCC scFv on the HCCcells. Methods: Retrovirus vector containing antiHCC scFv gene was transducted into HCC9204 cellsusing lipofectamine. Positive cell clones were selected with G418. The expression of scFv in HCC cellswas determined by RNA dot blot hybridization. Theproliferativity of transfected cells were assayed byflow cytometry. The morphologic changs were analyzed by light and electronic microscopy. Results:transfected HCC cells. Flow cytometric analysis oftransfected cells revealed the sub-G. DNA contentindicating the cell death. Light microscopy analysisshowed vacuolation in the scFv expressing cells andmore tumor giant cells apeared, these vacllolar organella were mitochondia and endoplasrnic reticulumwhen further studied by electronic microscopy. Inaddition, microvilli were reduced in scFv expressingcells. Conclusion: Intracellular expression of antiHCC scFv in HCC cells could induce the cell death.