Experiment To Study The Effect Of Colchicine On Rat C6 Glioma Cells In Vitro And In Vivo

Objective:1.To investigate the colchicine chemotherapeutic role on the proliferation of rat C6 glioma cells in vitro.2.To establish a reliable model of the rat brain glioma.3.To investigate the colchicine interstitial chemotherapy role on the general situation and survival time of the tumor-bearing rats in vivo.Methods:1.The rat C6 cells were divided into many colchicine treatment groups (2,1,0.5,0.25 ug·ml^-1 colchicine and so on),blank control group and zero group. When cells were cultivated to 1×10⁵/ml in the RPMI 1640 medium,they were inoculated into 96 holes board by 12 holes per group,100ul per hole to study.And MTT assay was used to observe the proliferation of glioma C6 ceils after incubated for 24h,48h and 72h.2.20 SD rats were randomly divided into test groups and control groups.In the experimental group,ten SD male rats were injected with 1×10⁶ C6 cells into the right caudate nucleus area using steoreotaxis techniques and in the control groups volume of serum-free culture medium of RPMI1640 were injected to observe the rate of rat brain glioma tumor.3.Thirty SD male rats were injected with 1×10⁶ C6 cells into the right caudate nucleus.At the 4th day after implantation the rats were divided into control group and experimental group.The colchicine group were treated with colchicine 10ul injected into the right caudate nucleus area using steoreotaxis techniques and the control group injected the same amount of saline in the same way for 3 times at the 4^th,7^thand10^th postoperation day to observe the colchicine interstitial chemotherapy role on the general situation and survival time of the tumor-beating rats.Results:1.MTT method in vitro experimental results showed that:colchicine could inhibit the growth of C6 glioma cells,the survived C6 glioma cells decreased with the increasing dose of colchicine and incubate time,which was time-dependent and dose-dependent effect.When the concentration of colchicine is 2ug/ml the growth inhibition rate reached 83.9 percent.The effects of colchicine on the proliferation of C6 glioma cells have a significant inhibitory effect.2.In the experimental group 9 rats died of glioma tumor within four weeks,one was long-term survival,the anatomy and pathological examination confirmed.The tumorigenic rate was 90%.3.In vivo experiments,the survival time of colchicine treated group was significantly prolonged than the saline treated group,there was a significant difference(P=0.000).Conclusion:1.Effects of colchicine on the proliferation of C6 glioma cells was inhibited,and with the increase in concentration and time,inhibition is also increased, which was a time-dependent and dose-dependent manner.2.By intracranial inoculation of 10⁶ rat C6 glioma cells to create the rat brain glioma model,the tumorigenic rate was 90%.The model is stable and reliable.3.The colchicine interstitial chemotherapy of the tumor-bearing rats can significantly prolong the survival time.4.Colchicine has the potential to become the new anti-tumor drug。...