The Study Of Antiproliferative Factor About The Proliferation Of Human Urinary Epithelial Cancer Cell Line T-24 And The Expression Of C-jun

Objective Antiproliferative Factor(APF) was discovered from interstitial cystitis patients by first.By chance,it had been discovered that it can inhibite bladder tumor cells growth.After APF affects normal urothelial cell,the C-JUN level decrease obviously,thereby it induce the whole level of Activator Protein-1(AP-1) decrease. AP-1 regulates the expression of target gene,include cell multiplication,cell differeneiation,inflammation,host response and canceration.And the activity loss of AP-1 affects the expression of the anti-oncogene P53 and cell cycle controlling gene P21(P21WAF1/CIP1) gene protein.The expression decrease of P53 and P21 gene restrain cell growth and proliferation,so cells show apoptosis or death.We can find different density APF and incompetence APF affect the cell growth of transitional cell carcinoma of the bladder by MTT and reverse transcription-polymerase chain reaction(RT-PCR).To approach the expression and signification of APF affecting transitional cell of the bladder.Method To cultivate bladder tumor cells T24 cell line.APF and incompetence APF according to different density divide to the expression group and control group,and each group affect T24 cells.Enzyme unite immunodetection equipment measure the extinction value of each group in OD 490nm by MTT,and to detect the cell growth rate of bladder tumor.The production was gotton by reverse transcription-polymerase chain reaction(RT-PCR) to make 2%agarose gel electrophoresis separation and EB dyeing showing.According to the electropherogram to get the width and brightness of strap,to carry out semiquantitative analysis to detect the expression of mRNA which is APF effect AP-1 after effecting bladder tumor cells.Result The result which is gotten by MTT measure the extinction value of each hole in OD 490nm by enzyme unite immunodetection equipment.It shows that to follow the increasing of APF density,the extinction value in OD 490nm degrade obviously,and the degrade of entinction value has statistical significance(P＜0.05);But the extinction value of adding incompetence APF has not marked change,and it has not statistical significance compare to experiment control group(P＞0.05).To measure the density of cDNA and OD260 and OD280 ratios by RT reaction,then regulate cDNA of each group to identical density,and carry out PCR reaction,to get the product making 2%agarose gel electrophoresis and EB dyeing.According to the electropherogram to get the width and brightness of strap,to carry out semiquantitative analysis.It shows that to follow the increasing of APF density,the expression of C-JUN is few and few. The difference has statistical significance among the different APF density groups.But it has not statistical significance between incompetence APF group and APF 0μg group.Conclusion APF may affect the expession of C-JUN which is a partment of AP-1. C-JUN affect cell-bridges directly,to carry to cell growth retardation.After APF affecting cells,the level of C-JUN degrade obviously,thereby cause to the whole level of AP-1 decrease and the degrade of it's activity affect the expression of the anti-oncogene P53 and cell cycle controlling gene P21 gene protein,so restrain the growth and proliferation of bladder tumor cells,and to follow the increasing of APF density,the depressant effect to cells is stronger and stronger.But inactive APF can not effect the expression of C-JUN,so it has not depressant effect to the growth of cells.It shows that APF which is applicated to treat the bladder tumor patients has very important value.The experimental result provide the basis for next animal experiment and clinical experiment.