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目的:将人增殖抑制基因 (hHSG)用电子穿孔的转基因方式转染到体外培养的人肿瘤细胞系中,观察hHSG基因对内源性hHSG表达水平不同的肿瘤细胞系的化疗敏感性的影响。方法:首先用免疫组化方法检测不同组织来源的肿瘤细胞系中hHSG的表达水平,然后选择内源性hHSG表达水平较低的肺腺癌 (A549 )和内源性hHSG表达水平较高的宫颈癌(HeLaS3)细胞系,用电穿孔方法转染含有hHSG的真核表达载体 (pEGFP hHSG) 24h后,加入放线菌酮(CHX),采用细胞计数、MTT法观察hHSG对肿瘤细胞增殖抑制作用及对CHX的化疗敏感性的影响。结果:hHSG在不同组织来源的肿瘤细胞系都有不同程度的表达, pEGFP hHSG转染到两种内源性hHSG表达水平不同的肿瘤细胞系后,这两种肿瘤细胞的生长增殖都明显受到抑制,同时外源性的hHSG也增强了这些肿瘤细胞系对CHX的敏感性。结论:外源性hHSG可不依赖其内源性表达水平而抑制肿瘤细胞的增殖,与CHX并用可增强肿瘤细胞对CHX的敏感性,具有协同作用。
Objective: To investigate the effect of hHSG gene on the chemosensitivity of tumor cell lines with different expression levels of endogenous hHSG by transfection of human proliferation suppressor gene (hHSG) by electron transfection into human tumor cell lines in vitro. Methods: Firstly, the expression of hHSG in tumor cells from different tissues was detected by immunohistochemistry. Then, the lung adenocarcinoma (A549) with low expression of endogenous hHSG and the cervix with high expression of endogenous hHSG (HeLaS3) cell lines were transfected with hHSG eukaryotic expression vector (pEGFP hHSG) by electroporation method 24h after adding cycloheximide (CHX), using cell counting, MTT assay hHSG inhibition of tumor cell proliferation And the impact of chemosensitivity to CHX. Results: hHSG was expressed in different degree in different tumor cell lines. The growth of both tumor cells was obviously inhibited after pEGFP hHSG was transfected into two tumor cell lines with different expression levels of hHSG , While exogenous hHSG also enhanced the sensitivity of these tumor cell lines to CHX. CONCLUSION: Exogenous hHSG can inhibit the proliferation of tumor cells independent of its endogenous expression level. Combined with CHX, it can enhance the sensitivity of tumor cells to CHX and has a synergistic effect.