摘要
目的 :研究细胞间间隙连接蛋白 (connexin43,Cx43)与基因治疗中旁观者效应的关系。方法 :将不同 Cx43表达水平的中国仓鼠卵巢细胞 (CHO)转染 tk基因 ,tk+ / tk- 细胞按不同比例混合 ,经更昔洛韦 (GCV )作用后 ,用细胞计数和流式细胞仪检测混合细胞体系生长抑制率。将上述混合细胞接种于裸鼠皮下 ,腹腔给 GCV 15 mg/ kg,每日 2次 ,3周后处死 ,观察皮下 CHO细胞结节的大小。结果 :体外实验证实 tk+ / tk- 混合细胞经 GCV作用后生长受抑制的程度与 Cx43表达水平有关 ,Cx43表达水平高的混合细胞在含有 5 % tk+ 细胞时就表现出旁观者效应 ,而 Cx43表达水平低的混合细胞在含有 2 0 % tk+ 细胞时才表现出明显的旁观者效应。实验发现 Cx43表达水平高的混合细胞动物皮下种植后经过 GCV作用 ,细胞结节的体积明显小于 Cx43表达水平低的混合细胞结节体积。结论 :在 HSV- tk-
Objective: To study the role of gap junction in bystander effect of gene therapy using HSV-tk-GCV system. Methods: Chinese hamster ovary (CHO) cell with different level of connexin43 expression were transfected with simplex virus thymidine kinase gene (HSV-tk). Different ratio of tk+/tk- cells in media were treated with 10 μmol/L GCV. The efficiency of bystander effect were evaluated by the inhibition of cell growth. Nude mouse were implanted subcutaneous with those mixed cell and treated with GCV 15 mg/kg twice a day. Three weeks later, the animals were sacrificed and the volume of subcutaneous mass were measured to assay the bystander efficiency in vivo . Results: In vitro study indicated that the inhibition of mixed cell system were correlated with the level of Cx43 expression. The inhibition of high Cx43 group were obvious when the ratio of tk+ cell was 5%, the same result did not appear until the ratio of tk+ cell was up to 20% in low Cx43 group. Study in vivo also showed the volume of subcutaneous mass were correlated with the level of Cx43 expression. The volume of high Cx43 group were small than the low Cx43 group. Conclusion: Gap junction-mediated intercellular coupling contributes to the bystander effect HSV-tk-GCV system to tumor.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
2001年第9期819-822,共4页
Academic Journal of Second Military Medical University
基金
国家自然科学基金资助项目 ( 39770 75 2 )
