摘要
目的 观察转TK基因肺癌细胞对丙氧鸟苷 (GCV)的敏感性及TK GCV系统杀灭肿瘤细胞机制。方法 观察转TK基因、转空载体A5 49细胞 (下分别简称A5 49 TK、A5 49 pLXSN)和A5 49细胞形态学改变、生长增殖特性。GCV对 3种细胞生长抑制 (MTT法 )。电镜、流式细胞仪观察A5 49 TK、A5 49细胞经 5 0μmol LGCV作用 3d后细胞凋亡。 结果 转基因细胞变为不规则 ,呈多角型 ,透亮度下降 ,体外增殖能力下降 ,A5 49 TK ,A5 49 pLXSN ,A5 493种细胞倍增时间分别为 42 .3 5、41.75、3 6.18h。依据IC5 0 ,A5 49 TK细胞对GCV敏感性比A5 49细胞提高 46倍 ,电镜发现A5 49 TK细胞有凋亡小体 ,核呈半月征等 ,流式细胞仪发现A5 49 TK、A5 49细胞亚G0 G1 期分别为 ( 12 .2 1± 1.76) %、( 1.3 2± 0 .47) % ,两者比较P <0 .0 1。结论 A5 49 TK细胞获得了对GCV敏感性 ,TK GCV杀灭肿瘤细胞与诱导凋亡有关。
Objective To observe the sensitivity of TK gene-transferred pulmonary carcinoma cells to GCV and study the mechanism of TK/GCV system killing carcinoma cells. Methods The morphism and reproduction competency of A549 cell, A549 cell transferred with TK gene (A549 TK) and A549 cell transferred with vacant vector (A549 pLXSN) were observed. The inhibitory effect of GCV on the 3 cells was studied with MTT assay. The apoptosis of A549 TK cell and A549 cell the treated with 50 μmol/L GCV for 3 days was observed with electron microscope and flow cytometer. Results Under the effect of GCV, A549 TK and A549 pLXSN cells became irregular, polyangular, and opaque. The reproduction competency decreased and the doubling time of cells of A549 TK, A549 pLXSN and A549 cell were 42.35, 41.75 and 36.18 h respectively, and the sensitivity of A549 TK cell to GCV was 46 times higher than that of A549 cell. Apoptotic bodies and semilunar features of cell nucleus were found under electron microscope. Flow cytometer revealed that G 0G 1 subphase of A549 TK, A549 cells were (12.21±1.76)%, (1.32±0.47)% respectively (P<0.01). Conclusion A540 TK cell have acquired sensitivity to GCV and TK/GCV killing carcinoma cell was relative to induced cell apoptosis.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2000年第9期872-874,共3页
Journal of Third Military Medical University
