Clinic And Experimental Study Of MDR Of Liver Cancer And Transcatheter Arterial Embolization/Infusion

To study the expression of P - glycoprotein( P - gp) and its effect on the response of trariscatheter arterial chemoembolization ( TACE) in patients with hep-atocellular carcinoma ( HCC ) , and investigate the feasibility of establishing tumor model of MDR in rabbits liver with VX2 tumor induced by adriamycin and the feasibility of MDR reversal.Material and Method1. Clinic trialTACE was performed in 47 consecutive untreated patients suffering from HCC with different doses of anticancer agent mixed with lipiodol according to their general condition and Child' staging. Routine dose of anticancer agent ( epirubicin or adriamycin 40 mg, mitomycin 20 mg, fliorouracil 500 mg, ) was used in 33 HCC patients and semidose was used in the other 14 patients. Biopsy was performed in all of the 47 patients prior to TACE with an 18 Gauge Biopsy needle under the monitor of fluoroscopy.Specimens were fixed in 10% neutral formaldehyde solution and embedded in paraffin; Serial sections were prepared at 4um from the samples for immuno-histochemical. P-gp expression was detected by Strept avidin ?biotin complex immunohistochemical technique using monoclonal antibody JSB - 1. Immunore-actions was observed under the light microscope and more than 10% positive cells was regard as P - gp expression. Follow up CT was performed 2 months after initial TACE. Tumor volume was calculated and the response to treatmentwas assessed according to the recommendation of the World Health Organization. Reduction of 50% or more in the total volume regarded as effective.2. Animal trial35 healthy (2. 5 -3.0 kg) and 2 VX2 tumor - carrying Japanese white rabbits were used in this study. 10 rabbits were used as carriers to maintain VX2 tumor and the other 25 were used to establish liver tumor. VX2 tumor was isolated from one of the tumor - carrying rabbits in sterile conditions under general anesthesia and cut into fine pieces in salt solution. Tumor suspension was injected into the thighs of rabbit to maintain the tumor. Chemotherapy with 4mg adri-amycin (diluted with 5ml salt solution) was performed once a week for three weeks via posterior auricular vein in another tumor carrying rabbit. VX2 tumor was surgically removed a week after chemotherapy and fresh tumor suspension was injected into the thighs of other two rabbits to maintain VX2 tumor. This process was repeated for three times. 25 white rabbits were randomly divided into five groups, five in each group. Laparotomy was performed through a mid - line abdominal incision under general anesthesia. A piece of 1mm3 VX2 tumor was implanted directly beneath the surface at the left side of the median hepatic lobe. For group A and B, the implanted tumor was isolated from no chemo -therapeutic induced tumor - carrying rabbits and for the other 15 rabbits in group CND and E the implanted tumor was surgically removed from the tumor carrying rabbits that experienced repeat chemotherapy. All of the 25 rabbits were underwent contrast enhance CT in the third week after inoculated into the liver to assessed the tumor volume. Different anticancer agents or saline was direct infused via the common hepatic artery of the five groups of rabbits under the condition of general anesthesia respectively. For group A and C 4 mg adriamycin ( dilute with 2ml of saline) was infused and for group B and D the same amount of saline was employed; For group E 1 mg of verapamil was infused before 4 mg adriamycin. Follow up CT were performed before the rabbits were sacrificed a week after treatment and compared with pretreatment to assess the change of the tumor for different groups. Specimens from the edge of tumor were stored under -70C, fixed in 10% neutral formaldehyde solution and 2.5% Glutaral solution respec-lively. Tumor structure was observed under microscope and JEM - 200EX transmission electron microscope. Single VX2 tumor cells suspension were made from tumor specimens stored under - 701. Apoptosis rate was analysis for each group by means of Flow Cytometry.3. Statistical analysis...