基因转移FasL诱导人黑色素瘤细胞凋亡的体外研究

目的：探讨体外基因转移Fas配体(FasLigand, FasL)对恶性人黑色素瘤细胞凋亡的影响。方法：用携带人FasL cDNA的缺陷型重组腺病毒载体(AdFL)，在体外转导2株黑色素瘤细胞，并使其表达；通过流式细胞仪、RTPCR法进行 Fas/FasL表达检测，TUNEL法及荧光显微镜检测细胞凋亡状况。结果：流式细胞仪和 RTPCR检测两株黑色素瘤细胞表面均表达Fas，不表达FasL，而AdFL转导的两株黑色素瘤细胞均能表达FasL；AdFL能显著诱导两株黑色素瘤细胞在体外凋亡或抑制其生长。结论：重组腺病毒FasL在体外诱导人黑色素瘤细胞凋亡效果显著。

Induction of Apoptosis in Melanoma by FasL Gene Transfer in vitro

Objective:To evaluate effects of Fas ligand (FasL) gene transfer in vitro for apoptosis of malignant melanoma. Methods:We constructed recombinant adenoviral vector containing human FasL cDNA(Ad-FL) under the control of a cytomegalovirus (CMV) promoter, which encodes human Fas ligand and transfected into two melanoma cells. Flow cytometric analysis, RT-PCR identify the expression of Fas/FasL. TUNEL, fluorescence microscope were used to analyse apoptosis. Results: Two melanoma cells surface express Fas, whereas not express FasL detected by RT-PCR and flow cytometric analysis; while tumor cells transducted by Ad-FL can express high level of FasL. Ad-FL can induce two melanoma cells apoptosis or suppress their growth in vitro. Conclusion: Recombinant adenovirus FasL had great effect on inducing the apoptosis of human melanoma in vitro.