导入FasL基因对荷肝癌裸鼠肿瘤增殖影响的实验研究

目的　对 PA 317/ Fas L - p L XSN在裸小鼠体内抗肿瘤作用及机理作进一步探讨。方法　制备 PA317/ p KXSN -Fas L +病毒上清及转 Fas L CBRH7919细胞株。 6 0只实验动物 ,随机分成 4组。收集处于对数生长期的 CBRH 7919肝癌细胞 ,A、C、D三组每只裸小鼠接种肝癌细胞于腋窝皮下。 A组 (对照组 ) :接种 5天后 ,在腋窝部肿瘤内注射生理盐水 ;B组 :接种Fas L - CBRH7919制备模型 ;C组 :实验方法同 A组 ,在腋窝部肿瘤内注射 PA 317/ p L XSN- Fas L +病毒上清 ;D组 :实验方法同 C组 ,在腋窝部肿瘤内注射 PA 317/ p L XSN空载体。肿瘤细胞接种后测量、计算肿瘤的体积 ;间接免疫荧光流式细胞仪(flowcytom eter,FCM)检测各组肿瘤组织 Fas L的表达情况。采用 PI- Annexin V方法 ,用 FCM检则各组肿瘤细胞凋亡情况。结果　观察时间为 2 0天 ,B、C组肿瘤生长受到明显抑制 ,但并未完全消退 ,且与对照组有显著差异 ;D组肿瘤生长与对照组无显著差异。 B、C组肿瘤组织中 Fas L表达显著高于 A、D组 (分别为 72 .3% ,6 8.6 % vs7.2 % ,11.3% ,P<0 .0 1) ;与此对应 ,B、C组肿瘤组织中细胞凋亡显著高于 A、D组 (分别为 30 .5 % ,31.3% vs 12 .1% ,19.4 ,P<0 .0 1)结论　采用逆转录病毒载体将Fas L基因导入裸小鼠肝癌组织 ,

THE EXPERIMENTAL STUDY ON THE EFFECT OF PROLIFERATION BY CONVERTING FASL GENE INTO HCC CELLS IN NUDE MICE MODEL

Objective In order to study the function and mechanism of PA317/FasL-pLXSN system to reject proliferation of hepatocellular carcinoma cells in unde mouse.Methods Recombinant retrovirus supernatant harvest and FasL-CBRH7919 cell line vwer prepared.Sixty unde mice were randomizedly divided into 4 groups.The CBRH7919 HCC cells was injected into armpit of each animal in A,C and D rgoup.A group(control group):every one received saline injected into the tumor tissue five day post inoculation;B group:FasL-CBRH7919 cells were injected into unde mice;C group:the treatment is similar to group A,saline was substituted with PA317/pLXSN-FasL supermatant harvest;Dgroup:the treatment is similar to group A,but saline was substituted with PA317/lLXSN dummy vector.The time-volume of tumors curve was drawn.The expression of FasL and apoptosis in tumor cells were detected by different methods with flow cytometer(FCM).Results The proliferation of tumor in group B and was restrained markedly than that of group A and D(P<0.01),but no regressed tumor was found.Correspondingly,the expression of FasL(72.3%,68.6% vs 7.2%,11.3%,respectively.P<0.01)and apoptosis(30.5%,31.3% vs 12.1%,19.4%,respectively.P<0.01) in tumor cells detected by FCM in group B and C were higher than that of group A and D.Conclusion It is an effective treatment on HCC by converting FasL gene into tumor tissue with retroviral vectors in nude mouse model,which could induce apoptosis and restrain proliferation of tumor,change the disproportion of FasL expression on the normal liver tissue,HCC tissue and the margins of the cancerous tissue.