基因转移FasL诱导人黑色素瘤细胞凋亡的体外实验

目的 探讨体外基因转移Fas配体（Fas-ligand,FasL)对人恶性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)，在体外转导两株黑色素瘤细胞，并使其表达；通过流式细胞仪，RT－PCR法进行Fas/FasL表达检测，TUNEL法及荧显微镜相关凋亡检测，分析，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凋亡效果显著。

Experimental Study of Gene Transfer FasL Inducing Apoptiosis of Melanoma Cells in Vitro

Objective To evaluate effects of gene transferring Fas ligand(FasL) in virtro on apoptosis of malignant melanoma.Methods They constructed recombinant advenoviral vector containing human FasL cDNA(Ad-FL) under the control of a cytomegalovirus (CMV) promter, which encoded human FasL and transfected it into two melanoma cells.The expression of Fas/FasL was identified by flow cytometric analysis and RT-PCR. TUNEL and fluorescence microscope were used to analyze apoptosis.Results Two melanoma cell surface expressed Fas, whereas did not express FasL that was found by RT-PCR detection and flow cytometric analysis; while tumor cells transducted by Ad-FL was able to express high level of FasL. Ad-FL was able to induce two melanoma cells apoptosis or suppress its growth in vitro.Conclusion Recombinant adenovirus FasL has great effect on inducing the apoptosis of human melanoma in vitro.