肿瘤抗原诱导T淋巴细胞协同丝裂霉素对膀胱肿瘤细胞株杀伤作用的研究

Study on the cytotoxic T-lymphocytes induced by soluble tumor antigen with cytotoxicity of mitomycin C against bladder carcinoma cell line TBC-1

①目的 研究膀胱肿瘤可溶性抗原诱导产生抗肿瘤特异效应细胞(BT-BAK)联合丝裂霉素对膀胱肿瘤细胞的杀伤机制 和衔接使用时机,探讨化疗和生物治疗有机结合的最佳方案。②方法 以BCG为佐剂、IL-2诱导健康人外周单核细胞产生BT- BAK细胞;丝裂霉素作用膀胱肿瘤细胞株2小时后观察TBC-1不同时间点HSP70的表达情况;TBC-1经丝裂霉素化疗2小时后各 个时间点加入BT-BAK细胞,比较不同时间点丝裂霉素和BT-BAK细胞衔接所产生的协同作用。③结果 丝裂霉素化疗TBC-1 后不同时间点0、8、24、48、72小时的HSP70的表达情况,OD值分别为0.306±0.005、0.255±0.009、0.203±0.018、0.237±0.029、 0.239±0.020。正常TBC-1的HSP70表达OD值为0.247±0.022。丝裂霉素化疗TBC-1后不同时间点0、8、24小时加入BT- BAK细胞,其杀伤活性分别为77.81%±3.14%、72.09%±3.20%、64.94%±4.12%。单纯丝裂霉素化疗组其杀伤活性为63.18% ±4.62%。④结论 TBC-1经丝裂霉素化疗诱导2小时后HSP70的表达增强,而且有一定的表达规律。HSP70的高表达有助于BT -BAK细胞对膀胱肿瘤细胞株TBC-1的识别,提高杀伤活性。丝裂霉素化疗TBC-1两小时后0～8小时点加入BT-BAT细胞可 以发挥两者的协同作用。

Objective To investigate the cytotoxicity of Mitomycin C with BT-BAK cells against human bladder carcinoma cell line TBC-1. Methods Peripheral blood mononuclear cells(PBMCs) were obtained from the healthy human donors, then stimulated by incubation with viable BCG and IL-2. The expression of HSP70 on TBC-1 were monitored at different time points after coincubation with mitomycin C for 2 hours, thereafter the BT-ABK cells were added at 0h、8h、24h time points and the MTT assay was used to measure cytotoxicity of Mitomycin C with BT-BAK against TBC-1.Thereafter the BT-BAK cells were added at 0h、8h、24h time points and the MTT assay was used to measure cytotoxicity of mitomycin C with BT-BAK against TBC-1. Results After coincubation with mitomycin C for 2 hours, the expression of HSP70 on TBC-1 were 0.306±0.005、0.255±0.009、0.203±0.018、0.237±0.029、0.239±0.020 at 0h、8h、24h、48h、72h. The normal TBC-1 was 0.247±0.022. The cytotoxicity of mitomycin C with BT-BAK against TBC-1 were 77.81%±3.14%、72.09%±3.20%、64.94%±4.12%. Conclusion The expression of HSP70 on TBC-1 may be increased after coincubation with mitomycin C for 2 hours. 0h-8h may be the best time for the TBC-1 to be killed by the BT-BAK cells after coincubation with Mitomycin C for 2 hours.It may be a simple way for mitomycin C with BT-BAK cells to kill the human bladder carcinoma cell line TBC-1,which may be used in hospital in the future.