CAJ | 학술논문

用梯形DNA电泳与Gimsa染色研究人T细胞瘤细胞株Jurkat在转导编码人6A8 α-甘露糖苷酶基因的反向cDNA后，对抗Fas 抗体诱导凋亡敏感性的变化，并用间接免疫荧光染色（流式细胞仪）检测细胞表面Fas分子的表达。结果表明，抗Fas抗体诱导细胞24h后，野生型及转导空载载体的Jurkat细胞出现明显的梯形DNA，两者之间无明显差异，但转导反向6A8 cDNA的细胞未见明显的梯形DNA。Gimsa染色结果显示转导反向6A8 cDNA的Jurkat细胞中凋亡细胞数明显减少，而转导空载载体则无影响。Jurkat细胞为Fas（CD95, Apo-1）全阳性细胞，转导反向6A8 cDNA或空载载体对Fas表达阳性率与Fas表达强度无明显影响。以上结果提示，转导编码6A8α-甘露糖苷酶基因的反向cDNA使人T细胞株Jurkat对抗人Fas抗体诱导的凋亡作用产生耐受。
용제형DNA전영여Gimsa염색연구인T세포류세포주Jurkat재전도편마인6A8 α-감로당감매기인적반향cDNA후，대항Fas 항체유도조망민감성적변화，병용간접면역형광염색（류식세포의）검측세포표면Fas분자적표체。결과표명，항Fas항체유도세포24h후，야생형급전도공재재체적Jurkat세포출현명현적제형DNA，량자지간무명현차이，단전도반향6A8 cDNA적세포미견명현적제형DNA。Gimsa염색결과현시전도반향6A8 cDNA적Jurkat세포중조망세포수명현감소，이전도공재재체칙무영향。Jurkat세포위Fas（CD95, Apo-1）전양성세포，전도반향6A8 cDNA혹공재재체대Fas표체양성솔여Fas표체강도무명현영향。이상결과제시，전도편마6A8α-감로당감매기인적반향cDNA사인T세포주Jurkat대항인Fas항체유도적조망작용산생내수。
DNA ladder and Giemsa's staining were used to detect changes of susceptibility of the Jurkat cells transducted with rAAV-reverse 6A8 α-mannosidase cDNA to apoptosis induction with mAb anti-Fas.Indirect immunofluorescent staining was performed to detect the Fas expression on cell surface.DNA ladder was observed in wild type and mock-transducted Jurkat cells,but not in the rAAV-reverse 6A8 cDNA-transducted ones after 24 h treatment with mAb anti-Fas.Giemsa's staining showed a number of dying and died cells (with apoptosis bodies) in wild type and the mock-transducted ones upon anti-Fas antibody treatment.Mock transduction did not affect the apoptosis induction.However,in the rAAV-reverse 6A8 cDNA-transducted cells,few apoptotic cells could be found.100% cells in each group expressed Fas molecular.The data indicated that Jurkat cells became resistant to apoptosis induction by anti-Fas antibody upon transduction with a reverse cDNA encoding human 6A8 α-mannosidase and the transduction did not affect the expression of Fas molecular.