A Preliminary Research On Apoptosis And Adhesion Of Bladder Carcinoma Cell Line T24 Induced By Chemokine Receptor CXCR3 Antagonist

Objective: We made a primary research on apoptosis and adhesion of bladder carcinoma cell line T24 induced by chemokine receptor CXCR3 antagonist. To explore the role of CXCR3/CXCL10 axis in bladder cancer cell growth and invasion and possibility of blocking this reaction axis by CXCR3 antagonists as a new target in the treatment of bladder cancer. Adhesion was considered as one initial step of cancer cell invasion. By examining the role of CXCR3 antagonists on T24 cell adhesion rate,we discussed its effects on cancer cell invasion. According to the results of the experiment, we analyzed and studied the possible mechanism of CXCR3 antagonist on T24 cell apoptosis and adhesion.Methods: Bladder carcinoma cell line T24 were cultured in vitro and we took cells in logarithmic phase as research objects. According to the requirement, we set up six groups in which concentration of CXCR3 antagonist(NBI-74330)was different as follows:0umol/L group(blank group)、20 umol/L DMSO group(control group)、5umol/L group、10 umol/L group、15 umol/L group and 20umol/L group. Apoptosis rates of each group on 24 h and 48 h were measured by flow cytometry(FCM),repeating three times. We focused on the two following problems: the first one was how the apoptosis rate of T24 cell changed with the increase of drug concentration;the second one was at the same concentration of CXCR3 antagonist, apoptosis rates on 24 h and 48 h of T24 cell were compared. Furthermore, we set up five groups(0umol/L(control group)、5umol/L、10 umol/L、15 umol/L、20 umol/L) to examine the adhesion rate of T24 cell in strict accordance with the cell adhesion experiment steps. MTT was used to test the adhesion rate of each group, repeating three times.Results: ⑴Effects on apoptosis rates of T24 lines Compared with DMSO(20umol/L) group, there were no difference in apoptosis rates of blank group and 5umol/L group(P>0.05);However, apoptosis rates of 10 umol/L、15 umol/L and20umol/L group increased as the impovement of concentration of CXCR3 antagonist and the following were the rates on 24 h and 48 h :(8.6+0.5)%、(14.7+0.4)%、( 18.5+0.5)% and(10.7+1.3)%、(16.1+0.4)%、(21.1+1.0)%,and clear difference was found between them(P<0.05). Moreover, Apoptosis rates of 15 umol/L group and 20umol/L group on 48 h were higher than that on 24h:(16.1+0.4)% vs(14.7+0.4)%and(21.1+1.0)% vs(18.5+0.5)%, furthermore, there was clear difference between them( P<0.05).However, no difference were found in other groups(P>0.05).⑵Effects on adhesion of T24 lines Compared with 0 umol/L(control group),adhesion rates of 5umol/L 、 10umol/L 、 15umol/L and 20umol/L decreased with different degrees,(94.2+1.0)%、(84.6+0.6)%、(74.9+1.6)%、(67.2+1.1)%,with statistical significance(P<0.05).Conclusions:1. CXCR3 antagonist showed some apoptosis-promoting effect on bladder carcinoma cell line T24, moreover, time dependence and concentration dependence had been found.2. Besides, It could also lowering the efficiency of cell adhesion in a dose-dependent manner which suggest that CXCR3 antagonist may somewhat inhibit T24 cell invasion.3. Blocking CXCR3/CXCL10 axis responses by CXCR3 antagonist may become a new target in bladder tumor therapy and provide new insights into treatment of bladder cancer.