Experimental And Clinical Studies Of Bleomycin On Craniopharyngiomas--Inhibitory Effect In Vitro And Intracystic Injective Therapy Effect

Craniopharyngioma, one of the most common congenital intracranialneoplasma, is very difficult to be resected for its deep site and theimportant structures adjacent. No measurement is recognized as ideal to date.It was said that bleomycin, which is repeatedly injected into the cysts,could annihilate the cystic craniopharyngiomas without any importantcomplication during one course. The foresight of this therapy seemspromising. However, few basic research on it has been reported, because thereis no constructed model both in vitro and in vivo. Based on these, we plan toprimarily culture its tissue to construct its model in vitro to study theantitumor effect of bleomycycin on it. The clinical effect of this drug isfurther determined on the campared outcomes of a group of control therapies.0bjective To investigate the experimental and clinical antitumor effect ofbleomycin on craniopharyngiomas and to search the theoretic data for itsclinical applying. To explore its antiterer mechanism by analysis thecomponent of the cystic fluids before and after chemotherapy with molecularbiological methods. Methods First Part:The dispersed cells were obtained fromthe fresh removed craniopharyngiomas and the cystic fluids through digestingin collagenase and washing with serum free EMEM respectively. Then they werecultured in vitro. The growing curve-lines was drawn on viable cell nunber incertain time point. In drug-sensitivity experiment, the medium of some wellgrowing cells were replaced by the bleomycin containing EMEM and the culturecontinued. The antitumor curve-line were also drawn by viable cell number insuccessive time point. In 144 hours some cells in the drug solution wereselected to assess the antitumor effect of bleomycin by ATP-luminescenceassay. Second Part: Some cystic craniopharyngiomas were mono-blindly andrandomly divided into 3 groups:a group - intracystic chemotherapy withbleomycin:b group - intracystic chemo-radiotherapy with bleomycin and ³²P; cgroup-intracystic radiotherapy with ³²P. The bleomycin, bleomycin with ³²P and0. 9% saline with ³²P were respectively injected into the cysts through thesilica tubes, which were set into the cysts in stereotactic operation. Thefollowing time sustained more than 6 months. The outcome of therapies weredetermined on comparing the injected cysts volume before treatment and afterfollowing up. The index and LD of the cystic fluids, blood and cerebral spinalfluids and the endocrine function of these patients were detected before andafter therapy. The effect of bleomycin on these factors were assessed onthese data.Third Part:The cystic fluids, cerebral spinal fluids and serums ofpatients from a and b groups were collected to extract proteins, which werethen quantified. Both the antigen of β-hcg and bcl-2 were detected in thesesamples by Western-blotting. The total RNA were isolated by Trizol methodsfrom some well growing cells, which were cultured in the EMEM containing 5%charcoal strapped FBS for 24hours. Both mRNA of D -hcg and bcl-2 were assayedWith Rt--PCR. Resu1ts First Part:Though al1 of the sentimental cells from thecystic fluids fai1ed to be cu1tured for 72hours, 4 fresh craniopharyngiomastissues had been successfu11y cu1tured for more than 2 weeks. The cel1s numberwas downtrend. The decreased speed related to the drug concentration. Theobvious antitumor effects were observed in lng/m1 and 0. 1mp/m1 groups, whi1ethe resistant appeared in 0. 01mg/m1 groups. Second Part: l4 patients hadfinished the whole therapic course, inc1uding 3 from a group, 6 from b and 5from c. Two patients were multicystic in a group and the drug was se1ectivelyinjected into the 1argest cyst. After fo1lowed up, the vo1umes of the drug-received cysts regressed 99% and 56. 5% respectively,while the drug--free cystsenlarged by multi times.The other monocyst in a group regressed 53. 9%. In bgroup, 3 cysts a1most disapeared, other 2 regress...