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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- GCV系统基因治疗过程中旁观者效应的效率和间隙连接蛋白的表达水平有关
Objective: To study the relationship between connexin43 (Cx43) and bystander effect in gene therapy. METHODS: The tk gene was transfected with different Cx43-expressing levels of Chinese hamster ovary cells (CHO). The tk+/tk- cells were mixed at different ratios and treated with ganciclovir (GCV) before cell count and flow cytometry. Mixed cell system growth inhibition rate. The above mixed cells were inoculated subcutaneously into nude mice and intraperitoneally given GCV 15 mg/kg twice daily for 3 weeks. The size of subcutaneous CHO cell nodules was observed. RESULTS: In vitro experiments confirmed that the degree of inhibition of growth of the tk+/tk- mixed cells after GCV treatment was related to the expression level of Cx43, mixed cells with high Cx43 expression exhibited bystander effect when containing 5% tk+ cells, and Cx43 expression. Low-level mixed cells exhibited a significant bystander effect when they contained 20% tk+ cells. It was found that the mixed cell animal with high expression level of Cx43 undergoes GCV after subcutaneous implantation, and the volume of cell nodule is significantly smaller than the mixed cell nodule volume with low Cx43 expression level. Conclusion: The efficiency of bystander effect and the expression level of gap junction protein in gene therapy of HSV-tk-GCV system