摘要
目的 探讨重组腺病毒mIkBα基因（AdmIkBα）联合γ射线对人高转移性肝细胞癌HCC9204细胞和裸鼠HCC9204移植瘤生长的影响。方法利用有限稀释法测定病毒滴度，以不同感染复数（MOI）的AdIxBαm（10、20、30、50）转染HCC9204细胞，Westemblotting法检测转染前后细胞1：）65表达和mIxBa表达。采用4Gyγ射线对各组细胞进行治疗。MTT法检测转染组、转染组＋放射组、对照组细胞生长抑制率。建立裸鼠HCC9204肝癌动物模型，肿瘤局部注射AdmIkBα30MOI并联合6Gyy射线治疗，在不同时间测量肿瘤体积。结果病毒滴度1．252×10^9pfu／ml。mIkBa蛋白随AdmlteBa剂量增加而进行性增加，P65蛋白随AdmIkBα剂量的增加呈进行性递减。转染组体外细胞生长随AdmIkBα剂量的增加逐渐受抑制，经7射线照射后生长抑制更加明显。AdmIkBα转染和放射联合治疗后7～28d，裸鼠肿瘤生长速率明显低于单纯照射组、单纯转染治疗组。结论AdmIkBα转染联合Y射线治疗可增强对肝痛细胞杀灭作用．联合治疗明品优于单纯放疗组和单纯基因治疗组。
Objective To explore the effect of recombinant adnenovirus vector mediated mutant IkBα （mIkBα） combined with radiation on the hepatocarcinoma. Methods Limited dilution method was used to test the virus titer in 293 cells. The HCC9204 cells were infected with MOI 10,20,30 and 50 for 48 h, respectively. The expression of p65 and mkB a protein was analyzed by Western blot. Transfected HCC9204 cells and controls were treated with 4 Gy T rays. The inhibition rate of HCC9204 cells was examined by MTF. Rat modlles of HCC9204 was constructed. AdmIkBα plasmids were injected into tumor tissue and the tumors were administered with 6 Gy T irradiation 48 hours later. Tumor growth at diferent time points was recorded during 28 days. Results The titer of AdmIkBα is 1.252 ×10^9 pfu/ml. The expression of mItcB protein was increased with titer of AdmIkBα, and p65 protein began to decrease when MOI was 10,and reached the lowest when MOI was 50, they were all dose-dependent. The proliferation of HCC9204 cell lines were suppressed, as was more significant combined with radiation, and the effect was in a viral dose-dependent manner. From days 7 to 28 after AdmIkBα gene and radiotherapy, the tumor growth was significantly slower than after irradiation or gene therapy alone. Conclusions Recombimant adenoviral-mediated mIkBα gene, combined with irradiation, can increase the cellkilling effect. It is better than that of either one alone.
出处
《中华放射医学与防护杂志》
CAS
CSCD
北大核心
2007年第5期454-457,共4页
Chinese Journal of Radiological Medicine and Protection
