Expression Of Apoptosis Inhibitor Gene Livin In Bladder Cancer Cell Lines

The latest reserch from both home and abroad about livin the newly found IAP family member has indicated that livin gene expressed highly in many malignant tumors.It plays a significant role in both occurrence and develepment of many kind of carcinomas.In our previous reserch we have demonstrated that livin gene expressed highly in bladder carcinoma which indicates that livin might play an important role in bladder cancer's occurrence and develepment.Objective:We choose two bladder cancer cell lines EJ and BIU-87,detecting the expression of livin gene in both of the two cell lines in order to further demonstrate the relations between livin and bladder cancer. Compareing the expression of livin in the cell lines,sieve one cell line for the consequent reserch by using molecular biologycal technique to interfere the livin gene in the bladder cell line.Methods:Human bladder cell lines BIU-87 and EJ are cultured in DMEM(Dulbecco's Modified Eagle's Medium)(1) RT-PCR is used to detect the livin mRNA in both of the two cell lines,and compare the transcription of livin gene in the tow cell lines.(2) ICC(Immunocytochemistry) is used to detect the translation of livin in the two cell lines,compareing the translation of livin gene,then analyse the resultes of RT-PCR and ICC,compare the expression of livin gene in EJ and BIU-87 cell lines.Results: Livin mRNA RT-PCR results suggest that the two cell lines have different levels of expression.Electrophoresis of the PCR product compared with the two cell lines can be found that EJ cell line has slightly stronger expression . The result of ICC indicates that two cell lines were both positive,which is according to the RT-PCR result. The results ofRT-PCR and ICC demonstrate that livin gene expresses highly in the two cell lines.Comparing the results,we can see that there is much higher expression in EJ cell line than that of BIU-87 cell line.Conclusions: In our previous research, RT-PCR was used to determine the expression of livin in 36 cases(31 men and 12 women) of TCC and 12 cases of normal bladder tissues (controls).LivinmRNA was expressed in 28 of 36 TCC cases (77.8%). Livin was no correlated with the grade, stage, quantity and recurrence. There was negative in the control group. The positive rate of livin between TCC and control was significant. In this paper,we experiment in two kinds of bladder cancer cells in vitro further. Immunocytochemistry and RT-PCR results show that livin expresses in two bladder cancer cell lines,this demonstrates that livin may play an important role in the metabolism and development of bladder cancer as well as bladder cancer chemotherapy resistance. Livin may become a new tumor marker in bladder cancer.By compareing the results of RT-PCR and ICC,EJ cell line is an ideal alternertive for further reserch .It is indicated that livin may play a significant role in the occurrence and development of TCC. Livin might be a biomarker for the detection of TCC and served as a new target for apoptosis-inducing therapy of TCC.