Sun Er-lin, Fan Xiao-dong, Han Rui-fa, Niu Yuan-jie
Tianjin Urology Institute 300211, China.
Zhonghua Zhong Liu Za Zhi. 2010 Apr;32(4):244-8.
To investigate the antitumor effect of recombinant IFN-alpha-2b-BCG on mouse bladder cancer MB49 cells in vitro, and to explore its antitumor mechanisms.
MB49 cells were co-cultured with recombinant BCG or wild BCG, and than were examined by light and transmission electron microscopy. The cell growth was assessed by MTT assay, and apoptosis rate and MHC-I of the MB49 cells was detected by flow cytometry using AO and Hoechst33258 fluorescence immunostaining.
The hIFN-alpha-2b-BCG-treated tumor cells showed slow growth, detachment of some cells, and various degree of degeneration. Light microscopy revealed organelle disorganization, chromatin aggregation, nuclear pyknosis, and cytolysis in some cells. Cellular membrane bulged and some bubbles were seen under fluorescence microscope using AO staining. Hoechst33258 assay also depicted frequent apoptosis in the tumor cells. The MTT assay showed that rBCG more actively than the wild BCG inhibited the proliferation of MB49 cells. The apoptosis rate of the recombinant BCG group was 19.7% and 46.6% at the time point of 24 h and 48 h, respectively, significantly higher than 10.8% and 20.9%, respectively, in the wild BCG group. The results of flow cytometry indicated that both types of BCG enhanced the expression of MHC-I in the MB49 cells, but more effective in the recombinant BCG group.
The recombinant hIFN-alpha-2b-BCG has more strong immuno-modulatory properties, anti-tumor effect on MB49 cells and induces apparent cytotoxicity in the bladder cancer cells in vitro.
研究重组干扰素α-2b-卡介苗对小鼠膀胱癌MB49细胞的体外抗肿瘤作用，并探讨其抗肿瘤机制。
将MB49细胞与重组卡介苗或野生卡介苗共培养，然后通过光学显微镜和透射电子显微镜进行观察。采用MTT法评估细胞生长情况，使用AO和Hoechst33258荧光免疫染色通过流式细胞术检测MB49细胞的凋亡率和MHC-I。
经hIFN-α-2b-卡介苗处理的肿瘤细胞生长缓慢，部分细胞脱落，并出现不同程度的变性。光学显微镜显示一些细胞的细胞器紊乱、染色质聚集、核固缩和细胞溶解。使用AO染色在荧光显微镜下可见细胞膜鼓起并出现一些气泡。Hoechst33258检测也显示肿瘤细胞中频繁发生凋亡。MTT试验表明，重组卡介苗比野生卡介苗更能有效抑制MB49细胞的增殖。重组卡介苗组在24小时和48小时时间点的凋亡率分别为19.7%和46.6%，显著高于野生卡介苗组的10.8%和20.9%。流式细胞术结果表明，两种类型的卡介苗均能增强MB49细胞中MHC-I的表达，但重组卡介苗组更有效。
重组hIFN-α-2b-卡介苗具有更强的免疫调节特性，对MB49细胞具有抗肿瘤作用，并在体外对膀胱癌细胞诱导明显的细胞毒性。
