摘要
目的 构建含 Fas L 基因的腺病毒载体 ,转染肾癌GRC- 1细胞株 ,观察 Fas L基因对 GRC- 1细胞株的影响 .方法 以腺病毒作载体 ,把 Fas L基因克隆入腺病毒载体 ,构建成含 Fas L 基因的转基因载体并酶切鉴定 ,用脂质体包裹法把含 Fas L 基因的腺病毒载体转染 GRC- 1细胞株 ,用 RT-PCR方法证实转染后有无 Fas L基因表达 ,采用电镜观察GRC- 1细胞形态学变化 .结果 酶切鉴定证实含 Fas L基因的转基因载体构建成功 ;RT- PCR显示转染后 GRC- 1细胞株Fas L 基因表达水平显著高于未转染细胞株 (P<0 .0 1) ;肾癌细胞株 GRC- 1转染 Fas L基因 72 h后 ,在光镜下可见细胞体积缩小 ,生长不良 ,染色质浓缩 ,密度增高 ,胞核中出现颗粒样物质 ,电镜下可见凋亡小体 .结论 成功构建了 Fas L 基因的转基因载体 ;转染 Fas L 基因可诱导肾癌 GRC-
AIM To construct adenovirus vector with FasL, transfer vector into GRC 1 cell line, and observe FasL effectson GRC 1. METHODS Using adenovirus as vector, cloned FasL gene into vector and identified if FasL gene was cloned into vector by using enzyme digestion; using Lipofectin to pack vector with FasL, transferred vector into GRC 1 cell line, and then observed morphological change of GRC 1 cell. RESULTS ① FasL gene was cloned into adenovirus vector by using enzyme cutting; ② After 72 hours after transferring FasL gene into GRC 1 cell line, FasL gene was higher expression level than untransferred cells; ③ Condensation of chromosome and apoptosis body were observed by electroscope. CONCLUSION Adenovirus Vector with FasL gene was sucessfully constructed; transferring FasL into GRC 1 cell may induced GRC 1 cell line to display cell apoptosis.
出处
《第四军医大学学报》
北大核心
2002年第11期973-975,共3页
Journal of the Fourth Military Medical University
