CAJ | 학술논문

目的观察腺病毒介导胸苷激酶/更昔洛韦(TK/GCV)系统联合肿瘤坏死因子-α(TNF-α)对鼠膀胱癌的杀伤作用.方法构建MB49小鼠皮下模型,随机分为对照组、TK/GCV组、TNF-α组、联合治疗组.按治疗计划分组进行病毒及药物注射,实验结束测量肿瘤体积大小,进行组织病理学检查.结果体外实验验证低浓度TNF-α联合GCV较单纯GCV对细胞的杀伤率明显增强,差异有统计学意义(P＜0.01).流式细胞仪检测,联合治疗组较单独用药组凋亡率明显提高,差异有统计学意义(P＜0.01).动物实验结束后肿瘤体积对比:TK/GCV组为(93.43±2.10) mm3、TNF-α组为(53.95±2.61)mm3、对照组为( 171.52±4.33) mm3、联合治疗组为(18.23±1.11) mm3,联合治疗组肿瘤体积较其他各组明显缩小,差异有统计学意义(P＜0.01).苏木素-伊红(HE)染色见各治疗组均出现细胞坏死凋亡,其中联合治疗组仅于周边见少量肿瘤细胞存活.结论 GCV对转染腺病毒MB49细胞及TNF-α对MB49细胞均有良好的抑制生长作用,且呈浓度依赖性；TK/GCV、TNF-α均能有效诱导凋亡,且两者具有协同作用；TK/GCV系统联合小剂量TNF-α能增强自杀基因系统对小鼠膀胱癌的治疗作用.
목적 관찰선병독개도흉감격매/경석락위(TK/GCV)계통연합종류배사인자-α(TNF-α)대서방광암적살상작용.방법 구건MB49소서피하모형,수궤분위대조조、TK/GCV조、TNF-α조、연합치료조.안치료계화분조진행병독급약물주사,실험결속측량종류체적대소,진행조직병이학검사.결과 체외실험험증저농도TNF-α연합GCV교단순GCV대세포적살상솔명현증강,차이유통계학의의(P＜0.01).류식세포의검측,연합치료조교단독용약조조망솔명현제고,차이유통계학의의(P＜0.01).동물실험결속후종류체적대비:TK/GCV조위(93.43±2.10) mm3、TNF-α조위(53.95±2.61)mm3、대조조위( 171.52±4.33) mm3、연합치료조위(18.23±1.11) mm3,연합치료조종류체적교기타각조명현축소,차이유통계학의의(P＜0.01).소목소-이홍(HE)염색견각치료조균출현세포배사조망,기중연합치료조부우주변견소량종류세포존활.결론 GCV대전염선병독MB49세포급TNF-α대MB49세포균유량호적억제생장작용,차정농도의뢰성；TK/GCV、TNF-α균능유효유도조망,차량자구유협동작용；TK/GCV계통연합소제량TNF-α능증강자살기인계통대소서방광암적치료작용.
Objective To investigate the killing effect of adenovirus-mediated thymidine kinase/ganciclovir (TK/GCV) gene therapy in combination with tumor necrosis factor-α (TNF-α) for murine bladder carcinoma cells.Methods The animal models were established,and divided into 4 groups randomly:control,TNF-α injected subcutaneously around the tumor,GCV injected intraperitioneally,TNF-α injected subcutaneously around the tumor + GCV injected intraperitioneally.The adenovirus and the drugs were injected as planned.During the course of the treatment,the volume of the tumors was measured.The tumor tissues were analyzed histopathologically.Results The survival rate of GCV on adenovirus-transfected MB49 cells decreased gradually as the concentration increased.The survival rate of TNF-α on MB49 cells decreased gradually as the concentration increased.When GCV in combination with low concentration of TNF-α were added,the killing rate of the infected murine bladder carcinoma cells in each group increased significantly (P ＜ 0.01 ).We can detected the apoptotic peak of sub-G1 stage in group of TK/GCV alone,TNF-α alone and TK/GCV in combination with TNF-α by flow cytometry.The apoptotic rate of the unification group increased significantly (P ＜ 0.01 ).When animal experimentin vivo completed,the average tumor volume of TK/GCV treated group was (93.43 ± 2.10) mm3,and TNF-α treated group was (53.95 ± 2.61 )mm3,While the average tumor volume of the control proup was ( 17 1.52 ± 4.33 )mm3.The average tumour volume of TK/GCV + TNF-α treatedgroup was ( 18.23 ± 1.11 )mm3,smaller than any others (P ＜ 0.01 ).Through histopathology detection we can see all the therapy group emerged cell necrosis and apoptosis.There are few tumor cell survived in bouncary in pathological section of the affiliation group.Conclusion GCV can inhibit the MB49 cells infected with TK gene efficiently.TNF-α can inhibit and kill the MB49 cells.There are dose-dependent relation.TK/GCV,TNF-α can effectively induce apoptosis in vitro,and have a synergistic effect.TK/GCV suicide genic system in combination with low doses of TNF-α can enhance the lethal effect of suicide gene on the murine bladder carcinoma MB49 cells.