Experimental Study Of3-bromopyruvate Targeting On Energy Metabolism Of Hepatic Carcinoma In Rats And The Assession Of Early Response To The Therapy

PART I:Experimental study of the antitumor effect of3-bromopyruvate targeting on Walker256cells in vitroPurpose:To assess the antitumor effect of3-bromopyruvate on Walker256cells.Materials and methods:MTT assay was applied to detect the antitumor activity of3-bromopyruvate on Walker256cells, cells apoptosis was analyzed using DNA ladder experiment in vitro.Results:The inhibition rate of Walker256cells treated with1,5,10,25,50umol/13-bromopyruvate were (10.9±0.9)%,(18.5±1.7)%,(24.2±1.3%,(40.4±3.0)%,(55.1±2.4)%, IC50of3-bromopyruvate on Walker256cells is40.26umol/L,72hours after treatment of50umol/L3-bromopyruvate on Walker256cells, DNA gel electrophoresis show apparent ladder, the negative control group had no ladder.Conclusions:Our results indicate that3-BrPA have a prospective antitumor effect in vitro. PART Ⅱ:Experimental study of3-bromopyruvate targeting on energy metabolism of hepatic carcinoma in rats through interventional procedures Purpose:To assess the antitumor effect of3-bromopyruvate on Walker256liver xenograft in Wistar rats model, and investigate its effect on tumor invasion and metastasis.Materials and methods:Walker256carcinona was implanted in the livers of rats to form tumor model. Rats in experimental groups (group A, n=10) were infused with3-bromopyruvate (1.5mg/kg concentration), rats in positive control group (group B, n=6) were infused mitomycin(0.1mg) and embolismed with lipiodol(0.1ml), rats in negative control group (group C, n=6) were infused with saline (1ml). After completion of the MRI examination, all the rats were sacrificed, the proportion of tumor necrosis was calculated based on histopathologic examination. Immunohistochemistry was used to detect tumors and adjacent normal tissue expression of VEGF and MMP9to evaluate the effects of3-bromopyruvate on metastasis of Walker256liver xenografts.Results:Tumor necrosis rate:7cases with grade Ⅲ necrosis,2cases with grade Ⅳ necrosis and1cases with grade Ⅰ necrosis in group A,6cases of group B were grade Ⅰ necrosis,5cases grade Ⅲ necrosis and1cases grade Ⅱ necrosis in group C, the mean tumor necrosis rate of group A was significantly higher than that of group B (P=0.001), groups A and C was no significant difference in mean tumor necrosis rate (P=0.927). Immunohistochemical examination showed VEGF of the tumor center in group A have9cases with negative expression and1case with grade Ⅱ expression, there are9cases with grade Ⅰ and1case with grade Ⅱ VEGF expression of adjacent tissues in group A; the VEGF of the tumor center in group B have4cases with negative expression and2cases with grade Ⅰ expression, there are5cases with grade Ⅱ and1case with grade Ⅰ VEGF expression of adjacent tissues in group B; the VEGF of the tumor center in group C have6cases with grade Ⅱ expression, there are6cases with grade Ⅱ VEGF expression of adjacent tissues in group C; VEGF expression of cancer center of in groups A and B was significantly different with group C (P=0.001, P=0.002), there was no statistically significant of VEGF expression in groups A and B (P=0.339), there was significant difference of VEGF expression in paracancerous in C group with group A, B (P =0.005, P=0.001), paracancerous VEGF was no significant difference in group B, C (P=0.317); MMP9expression of cancer centers were7cases with negative,1case with grade Ⅱ and2cases with grade Ⅰ in the group A, paracancerous expression of MMP9were8cases of grade Ⅰ and2cases negative in group A; MMP9expression of cancer centers were4cases with negative,2cases with grade Ⅰ in group B, MMP9expression of paracancerous were6cases of grade Ⅰ in group B; MMP9expression of cancer centers were5cases with grade Ⅱ,1case with grade Ⅰ in group C, paracancerous expression of MMP9were6cases of grade Ⅱ in group C; Expression of MMP9of cancer center was no significant difference in group A and B (P=1.000), Expression of MMP9of cancer center in group C was different from the group A and B (P=0.003, P=0.004); MMP9expression in tumor adjacent tissues was no significant difference between group A and B (P=0.257), MMP9expression in tumor adjacent tissues in group C was significantly different from group A and B(P=0.002, P=0.005).Conclusions:Our results indicate that intraarterial delivery of3-BrPA shows a promising antitumor effect in vivo. PART Ⅲ:The appliacation of MR-ADC in evaluating early response to the therapy targeting on hepatic carcinoma in rats by3-bromopyruvate through interventional proceduresPurpose:. To study of diffusion-weighted imaging in assession of the therapy targeting on energy metabolism of Walker256liver xenograft in Wistar rats by3-bromopyruvate through interventional procedures.Materials and methods:Walker256carcinona was implanted in the livers of rats to form tumor model. Rats in experimental groups (group A, n=10) were infused with 3-bromopyruvate (1.5mg/kg concentration), rats in positive control group (group B, n=6) were infused mitomycin(0.1mg) and embolismed with lipiodol(0.1ml), rats in negative control group(group C, n=6)were infused with saline (1ml). Maximum diameter,the mean volume and apparent diffusion coefficient (ADC) of hepatic carcinoma in the model rats were measured by MRI on the day before interventional treatment and the third day after interventional treatment.Results:Maximum diameter of tumor before interventional treatment was9.53±2.48mm,7.35±3.61mm and4.51±3.08mm in groups A,B and C, respectively; On the third day after treatment, the maximum diameter of tumor was15.20±2.76mm,10.48±2.01m and10.13±4.03mm in groups A,B and C, respectively; Groups A,B and C no significant difference in tumor maximum diameter (P=0.795, P=0.630, P=0.842). The mean tumor volume of tumor before treatment was305.48±62.47mm3,439.62±196.58mm3and137.93±93.93mm3in groups A,B and C, respectively, On the third day after3-bromopyruvate treatment, the mean tumor volume was482.98±103.88mm3,569.79±268.89mm3and432.44±437.39mm3in A,B and C groups, respectively; Groups A and B, C no significant difference in tumor volume(P=0.781, P=0.638); ADC values of tumor before interventional treatment was (8.37±1.77)×10-4s/mm2,(7.59±0.96)×10-4s/mm2and (6.97±1.51)×10-4s/mm2in groups A,B and C, respectively; On the third day after treatment, ADC values of tumor were (15.15±5.20)×10-4s/mm2,(8.23±1.12)×10-4s/mm2and (12.23±1.97)×10-4s/mm2in A,B and C groups, respectively; After treatment, ADC values of groups A and B were significantly different with group C (P=0.009, P=0.004), ADC values was no significant difference between A and group B (P=0.159).Conclusions:The ADC values of tumor can be applied to assess the early effects of3-bromopyruvate in rat hepatoma model.