The Research About The Killing Effect Of ¹²⁵IUdR On The CSCs Of The C6 Glioma Cell

OBJECTIVE:Try to understanding the effect of ¹²⁵IUdR on the brain tumor CSCs. Know whether this drug could inhabit the proliferation of brain tumor CSCs.METHORD:The experiment can be divided into two parts,the in vitro one and the in vivo one.1.As the in vitro one we use the Immunocytochemistry to determine CSCs exists in the C6 cell line(RFP Nestin antibody and GFP CD133 antibody culture with the C6 cell line for more then 24 hours,antibody dilute by 1:10000 and cover the whole surface of the cell culture tank).Then we use the flow cytomtery (backman)to separate the CSCs of C6 cell line.At last,co-culture the C6/CSCs with ¹²⁵IUdR(0.2mCi/10μl)and evaluate the killing effect of this medicine by flow cytometry after 24 hours.2.For the in vivo experiment we use the animal mode of C6 gliomas to evaluate the killing effect of ¹²⁵IUdR.Divided the C6/SD rats mode into three group every group contains 10 animals:the experiment group injects ¹²⁵IUdR at C6 gliomas local section for ¹²⁵IUdR(0.2mCi/10μl/8h),1time/2d,6days.;the control group injects physiological brine at the C6 gliomas local section for same quantity,1time/2d,6days;the blank group were killed derectly when the model is established.Then evaluate the kill effect through CSCs numbers by Nestin CD133 coimmunocytochemistry.RESULT:1.As the in vitro experiment show that the C6 cell line include CSCs which react with both antibody.2.The flow cytometry do separate the CSCs from the C6 cell line well.3.As the CSCs coculture with the ¹²⁵IUdR after 24 hours the flow cytometry assay show the killing effect well.4.The xenografty SD mouse really contain CSCs verified by HE and immunocytochemistry.5.Comparative between the experimental group and the control group by immunocytochemistry and Error bar,it shows the killing effect of ¹²⁵IUdR on the in vivo brain tumor.DISCUSS:1.As the in vitro experiment show the ¹²⁵IUdR really could kill the CSCs effectively in vitro.2.The ¹²⁵IUdR also work well in vivo.It kills the CSCs effectively in vivo.So the ¹²⁵IUdR killing effect on CSCs may lead to those tumor behavior restrained.