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目的观察干扰素γ(IFN-γ)与树突状细胞(DC)疫苗联合使用后对口腔鳞癌的抑制作用。方法采用RT-PCR方法分析抗原提呈相关基因Tap1和Tapasin在人正常口腔黏膜细胞以及口腔鳞癌细胞系中的表达。采用IFN-γ处理口腔鳞癌细胞系CAL27后,分别在mRNA和蛋白水平检测Tap1和Tapasin的变化。将处理后的肿瘤细胞提取冻融抗原后致敏DC,并将其与同源的T淋巴细胞共培养,诱导出抗原特异性的细胞毒T细胞(CTL),在体外对口腔鳞癌细胞进行杀伤试验。结果与正常人口腔黏膜上皮细胞相比,肿瘤细胞中Tap1和Tapasin的表达明显降低(均在50%以下)。使用IFN-γ处理后,其在mRNA和蛋白水平的表达均明显提高。CTL行体外杀伤实验时发现,IFN-γ处理组肿瘤杀伤效率明显提高,A组(IFN-γ处理48 h)的肿瘤细胞杀伤率达到(87.21±4.67)%,而B组(IFN-γ处理24 h)及C组(IFN-γ处理0 h)则分别达到(73.34±4.52)%和(54.68±4.21)%(P<0.01)。结论 口腔鳞癌细胞中抗原提呈相关基因Tap1和Tapasin的表达明显降低,使用IFN-γ处理后可明显提高其表达。将处理后的肿瘤细胞提取抗原后,诱导出抗原特异性CTL,可显著提高其肿瘤杀伤率。
Objective To observe the inhibitory effect of interferon γ (IFN-γ) combined with dendritic cell (DC) vaccine on oral squamous cell carcinoma. Methods The expression of Tap1 and Tapasin in human normal oral mucosa cells and oral squamous cell carcinoma cell lines were analyzed by RT-PCR. The IFN-γ-treated oral squamous cell carcinoma cell line CAL27 were detected at the mRNA and protein levels of Tap1 and Tapasin changes. The treated tumor cells were extracted after freezing and thawing antigens, and DCs were sensitized and co-cultured with homologous T lymphocytes to induce antigen-specific cytotoxic T cells (CTLs). The in vitro squamous cell carcinoma cells Kill test. Results Compared with normal human oral epithelial cells, the expression of Tap1 and Tapasin in tumor cells were significantly decreased (both below 50%). After treatment with IFN-γ, its mRNA and protein levels were significantly increased. CTL in vitro killing experiments found that IFN-γ-treated group significantly increased tumor killing efficiency, A group (48 h IFN-γ treatment of tumor cell killing rate of (87.21 ± 4.67)%, while the B group (73.34 ± 4.52)% and (54.68 ± 4.21)%, respectively (P <0.01) when compared with control group (24 h) and group C (0 h IFN-γ treatment) Conclusion The expression of Tap1 and Tapasin genes in oral squamous cell carcinoma tissues was significantly lower than that in control group. IFN-γ treatment could significantly increase the expression of Tap1 and Tapasin. After treating the tumor cells to extract the antigen, antigen-specific CTL can be induced, which can obviously increase the tumor killing rate.