Cholangiocarcinoma Cell Lines Qbc939 Nude Perineural Invasion Model And 5'-amp Transplanted Tumors In Nude Mice Cholangiocarcinoma Experimental Studies

Objective:To establish a model of cholgangiocarcinom perineural invasion in nude mice with Cholangiocarcinoma cell line QBC939. To investigate the effects and pathological changes of biological hypothermia caused by 5'-AMP on the growth of transplanted tumor of Cholangiocarcinoma QBC 939 in nude mice.Methods:①Human cholangiocarcinoma QBC 939 cells were anabiosissed, cultivated and passaged enough. And then QBC 939 cells were implanted subcutaneously into posterior portion of nude mice. Primary transplanted QBC 939 cell line of cholangiocarcinoma in nude mice was set up. Then part of primary transplanted tumour was transplanted into peritoneal cavities of 20 nude mice. After 30 days, passaged transplanted mice were killed to undergo anatomical and histopathological examination, meanwhile, perineural invasion around biliary tract of mice was observed.②Fourteen posterior portion tumorgenesis nude mice were separated into two groups randomly: The first group is experimental group, which is injected relevant dose of 5'-AMP in peritoneal cavity; The other one is blank group, which is injected equivalent normal sodium in peritoneal cavity. The next day, the mice were killed, (1) the tumor tissue underwent HE stained and proportion of tumor necrosis were observed, (2) the level of serum tumor marker, CEA and CA19-9 were tested.Results:1. All of five primary transplanted cholangiocarcinoma in nude mice were found tumorigeness, emerging rate of tumor is 100%(5/5); nine of twenty peritoneal cavities passaged transplanted nude mice were found tumorigeness, two of which were found perineural invasion around biliary tract; emerging rate of tumor is 45%(9/20), the rate of perineural invasion around biliary tract of tumor is 22.2%(2/9).2. The group of mice which were injected 5'-AMP in the dose of 3mg/g got into hibernation in 60 minutes in average when temperature fell down to 25℃; The temperature got the minimum-22.0℃,which lasted about 60minutes, then temperature rose gradually and returned to normal body temperature in about 350 minutes; In the next day all mice survived, the average measured temperature 36.3℃, with stable vital signs.3. Averaged percent of necrotic tissue of 5,-AMP experimental group is (18.2±5.3)%, averaged percent of necrotic tissue of blank control group is (5.7±2.4)%, The difference was significant between the experimental and control group in terms of averaged percent of necrotic tissue (P＜0.05). The mean values of CA19-9 and CEA in serum of experimental group were(318.6±102.9)kU/L and(11.9±0.8)μg/L, The averaged values of CA19-9 and CEA in serum of control group were (452.7±118.3) kU/L and (13.1±0.9)μg/L, The difference was significant between the experimental and control group in terms of averaged values of CA19-9 and CEA in serum (P＜0.05).Conclusion:1. The model of cholgangiocarcinom perineural invasion in nude mice were established, the success rate is 10%, it share same pathological characteristics with human cholangiocarcinoma.2.3mg/g is the optimum dose to induce biological hypothermia, and it gets the minimum in 120 minutes.3.Biological hypothermia caused by 5'-AMP shows apparent inhibition not only on the growth of transplanted cholangiocarcinoma in nude mice, but also on other tumors.