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第五届中国肿瘤学术大会二等奖--郑传胜论文摘要
2008-10-21 04:56  稿源:威尼斯城所有登入网址-威尼斯人平台网址-首页

  重组人P53腺病毒基因经介入方法治疗肝癌的的动物实验研究

  华中科技大学同济医学院附属协和医院介入科 郑传胜

     摘要 目的 研究经不同介入导入方式注射重组人P53腺病毒基因(rAd-p53)在兔VX2肝癌模型中的表达特点及治疗作用。材料与方法 (1)将27只种植成功的兔VX2肝癌模型随机分成三组(每组9只):A组:经肝动脉单纯灌注rAd-p53(5X106 VP);B组:rAd-p53(5X106 VP)+超液态碘油0.5ml经肝动脉混合灌注;C组:瘤内直接穿刺注射rAd-p53(5X106 VP)。术后24h、72h、6天分别处死实验兔,通过免疫组织化学方法检测肿瘤组织及瘤周P53 、Bax和Bcl-2蛋白的表达。(2)将30只兔VX2移植性肝癌模型随机分成五组(每组6只),a组:经肝动脉灌注生理盐水0.5ml;b组:经肝动脉灌注超液态碘油0.5ml行栓塞治疗;c组:经肝动脉单纯灌注rAd-p53(5X106 VP);d组:rAd-p53(5X106 VP)+超液态碘油0.5ml经肝动脉混合灌注;e组:瘤内直接穿刺注射rAd-p53(5X106 VP)。术后2周处死实验兔,先行大体病理检测,计算肿瘤生长率;再分别通过免疫组织化学方法检测肿瘤细胞凋亡、血管内皮生长因子(VEGF)及Von-Willebrand-factor(VIII因子)表达情况,对VIII因子阳性血管内皮细胞进行微血管(MVD)计数。比较分析不同导入方式的rAd-p53治疗作用。结果(1)rAd-P53基因的表达情况:A组,P53蛋白在24h、72h及6天平均表达率(%)分别为19.65±5.29, 62.43±3.27, 18.73±3.45;B组P53蛋白在24h、72h及6天平均表达率(%)分别为20.28±4.34, 65.43±2.13 ,19.63±4.23;C组P53蛋白在24h、72h及6天平均表达率(%)分别为26.53±6.07,70.36±3.43 ,26.36±4.35。P53和Bax蛋白的表达呈与时间相关性正态分布,以C组表达最高(P<0.05),Bcl-2蛋白表达随P53蛋白表达的增多而减少。(2) rAd-P53基因的治疗作用:c、d、e组肿瘤增长率较a、b组低(p<0.05),其中以d组最低(1.65倍)。五组(a、b、c、d、e)肿瘤细胞平均凋亡率分别12.06%、14.56%、17.65%、18.69%、19.65%,差异有显著性(P<0.05)。五组VEGF阳性率分别为50.0%、83.3%、83.3%、50.0%、50.0%,差异有显著性(P<0.05)。五组MVD计数分别为81.62±16.13 、85.23±24.34 、75.20±23.91 、71.16±21.34 、72.34±25.29,差异有显著性(P<0.05)。结论 (1)重组人P53腺病毒基因在兔VX2肝癌模型中的表达呈正态分布, rAd-p53不同导入方式,P53、Bax、Bcl-2蛋白表达的程度不同,以瘤内直接注射表达最高,与超液态碘油混合灌注表达次之。(2)经介入途径导入重组人P53腺病毒基因能安全高效促进兔VX2肝癌细胞凋亡、抑制肿瘤血管生成,进而抑制肝癌细胞转移;不同的rAd-p53导入方式对肝癌治疗效果不同,与超液态碘油混合灌注最为有效,瘤内直接注射次之。

  Experimental studies of recombinant human adenovirus P53 in the interventional treatment for VX2 rabbit transplanted hepatocarcinoma

  Objective: To evaluate the efficiency of recombinant human adenovirus P53(rad-P53) in the interventional treatment of rabbit VX2 heptocarcinoma model by different injection ways .methods: Thirty Newzealand rabbit VX2 hepatocarcinoma models were established .Then they were splited randomizedly into five groups. The therapy scheme performed as below: group A was infused through hepatic artery with 0.9%NaCl 0.5ml, group B with ultrofluid lipiodol 0.5ml, group C with rAd-p53(5×106VP) simply, group D with ultrofluid lipiodol 0.5ml + rAd-p53(5×106VP), group E was injected in tumors with rAd-p53(5×106VP) through surgery directly. They will be observed by pathological after death, then they were detected by stain of immunohistochemical method about the level of apoptosis cell and the expression of VEGF、von-willebrand-factor(VIII) after two weeks of the operation. MVD was counted by endothelial cells which were highlighted by factor VIII, their expression levels were analyze in conjunction with the pathological feature.Results: The volume and the ratio of growth were sinnificant difference after interventional therapy of all groups(p<0.05). The groups of C, D and E were lower than groups A and B, group D was the lowest. The mean apoptosis index(AI) are 12.06%、14.56%、17.65%、18.69% and 19.65% respectively(p<0.05). The mean positive ratio of expression of VEGF are 50.0%、83.3%、83.3%、50.0% and 50.0% respectively(p<0.05). The mean MVD expression of five groups are 81.62±16.13、85.23±24.34、75.20±23.91、71.16±21.34 、72.34±25.29 respectively (p<0.05).Conclusions: The rAd-p53 have the safe and high effect of promote VX2 rabbit hepatocarcinoma cells apoptosis and anti-angiogenesis and inhibiting growth and metastasis through interventional therapy. There are differences of therapeutic effect among the different methods of interventional guiding of rAd-p53, the method of injected with ultrofluid lipiodol + rAd-p53 hrough hepatic artery is the best.

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