| Part one Establishing rabbit VX2 hepatocarcinoma model and the features of expression of recombinant human adenovirus P53 injection in the model through interventional guidingObjective: To investigate the feasibility of establishing rabbit VX2 hepatocarcinoma model and study the features of expression of recombinant human adenovirus P53(rAd-p53 ,1X1012VP) injection in the model through interventional guiding.Materials and methods: Sixty Newzealand rabbits hepatocarcinoma model were established by VX2 tumor implantation, the tumor volume(V1) was measured by magnetic resonance imaging(MRI) and CT scanning 11 days after implantation. Twenty-seven rabbits were extracted randomizedlly after the successful implantation. Then they were divided into three groups. Group A was infused through hepatic artery with rAd-p53(5×106VP) and group B was ultrofluid lipiodol 0.5ml+rAd-p53(5×106VP) as well, then the group C was injected in tumors with rAd-p53(5×106VP) through surgery directly. All the rabbit model were performed DSA examination during operation. All the rabbits were detected by stain of immunohistochemical method about the expression of P53,Bax and Bcl-2 protein in the tumors and around tumors after the operation in 24h, 72h and 6 days. Result: Succecssful implantation was obtained in fifty-seven rabbits by the examination of MRI and CT scanning, the ratio of successful was 95%. They were low density or the same density in plain CT scanning , low signal in T1WI imging and high signal in T2WI imaging ,the signals were well-distributed and clear boundary in MRI 14 imaging. Through DSA examination, the supply of the tumor vessel mainly coming from rabbit hepatic artery. They were all rich in blood vessel and tumor stain were be founded. The mean expression of P53 protein of group A after 24h, 72h and 6 days were 19.65±5.29, 62.43±3.27, 18.73±3.45;The mean expression of P53 proein of group B after 24h, 72h and 6 days were 20.28±4.34,65.43±2.13 ,19.63±4.23;The mean expression of P53 protein of group c after 24h, 72h and 6 days were 26.53±6.07,70.36±3.43 ,26.36±4.35. The mean expression of Bax protein of group A after 24h, 72h and 6 days were 8.56±1.27 ,35.75±1.35 ,9.84±2.46;The mean expression of Bax protein of group B after 24h, 72h and 6 days were 11.32±2.13 ,37.63±2.46,10.34±2.13; The mean expression of Bax protein of group C after 24h, 72h and 6 days were 17.64±2.25 ,43.76±2.19 ,16.89±1.67. The mean expression of Bcl-2 protein of group A after 24h, 72h and 6 days were 30.43±3.06 ,18.03±4.36 ,29.65±4.32;The mean expression of Bcl-2 protein of group B after 24h, 72h and 6 days were 28.36±2.25 ,16.21±3.17 ,27.70±3.39;The mean expression of Bcl-2 protein of group C after 24h, 72h and 6 days were 24.27±4.37 ,12.07±2.64 ,23.84±2.51.The expression of P53 and Bax protein in the tumors and around the tumors were shown in normal distribution, and the low level of expression of Bcl-2 protein was associated with the high expression of P53 protein.Conclusion: Rabbit VX2 heptocarcinoma model is suitable for interventional experiment study on hepatocarcinoma because of it is similar with human hepatocarcinoma in growth styles. The expression of P53 protein in the model of Rabbit VX2 heptocarcinoma is shown in normal distribution and the expression of Bax,Bcl-2 protein are associated with P53 protein. There are differences among the three different methods of interventional guiding, the method of injected in tumors with rAd-p53 through surgery directly is the best, the method of infusing throuth hpatic artery is lower.Part two Experimental studies of recombinant human adenovirus P53 injection treatment for VX2 rabbit transplanted hepatocarcinoma through interventional therapyObjective: To evaluate the efficience of recombinant human adenovirus P53 injection in the treatment of rabbit VX2 heptocarcinoma model by different ways through interventional therapy.Materials and methods: Thirty Newzealand rabbits hepatocarcinoma model were established in part one , the tumor volume(V2) was measured by MRI and CT scanning 14 days after interventional operation to analysis the growth rate. All the rabbit model were performed DSA examination during operation. 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 was ultrofluid lipiodol 0.5ml; group C was infused through hepatic artery with rAd-p53(5×10~6VP) simply; group D was ultrofluid lipiodol 0.5ml + rAd-p53(5×10~6VP); group E was injected in tumors with rAd-p53(5×10~6VP) 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 interventional operation. MVD was counted by endothelial cells which were highlighted by factor VIII, their expression levels were analyze in conjunction with the pathological feature. Result: There is no difference among the volume before therapy(P>0.05), but 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. Significant difference exited in microxcopic metastasis foci of paracancer livers tissue among all groups, the group B was lower than groups A,C(p<0.05), but the group B and D was no difference(p>0.05), group E was higher than group D,C(p<0.05), group D was lower than group E(p<0.05) . The mean apoptosis index(AI) are 12.06%,14.56%,17.65%,18.69% and 19.65% respectively. The AI of group C,D and E are significant difference exited than group A and B (p<0.05) by statistical analyze, and are significant difference exited among group C,D and E(p<0.05), but not between C and D(p>0.05). The mean positive ratio of expression of VEGF are 50.0%,83.3%,83.3%,50.0% and 50.0% respectively, the group B is significant difference exited than group A(p<0.05)by statistical analysis, and are significant difference exited among group C,D and E(p<0.05), but not between C and D(p>0.05). The mean expression of five groups of MVD are 81.62±16.13,85.23±24.34,75.20±23.91,71.16±21.34,72.34±25.29 respectively, the group B is significant difference exited than group A(p<0.05), the group C,D and E are significant difference exited than group A and B (p<0.05) by statistical analyze, and are significant difference exited among group C,D and E(p<0.05), but not between D and E(p>0.05). Statistical analysis showed a positive correlation between the expression of VEGF and MVD.Conclusion: The rAd-p53 have the safe and high effect of promote VX2 rabbit hepatocarcinoma cells apoptosis and anti-angiogenesis and inhibiting growth and metastasis as well through interventional therapy. There are differences of therapeutic effect among the different methods of interventional guiding of rAd-p53, the method of injected in tumors through surgery directly plus conventional invterventional therapy is the best, injected in tumors through surgery directly and the method of infusing throuth hpatic artery is lower.
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