GM-CSF分泌型肝癌疫苗对移植性肝癌小鼠CTL杀伤活性的作用机制  

Research on mechanism of GM-CSF secreting liver cancer vaccine on CTL killing activity of transplanted liver cancer mice

作　　者：吴天山[1] 肖开提.伊布拉因 阿尔帕提.买买提 李相成[2] 

机构地区：[1]新疆医科大学第一附属医院急诊外科,新疆乌鲁木齐830054 [2]南京医科大学临床医学系,江苏南京210029

出　　处：《中国生化药物杂志》2015年第3期62-64,68,共4页Chinese Journal of Biochemical Pharmaceutics

基　　金：国家自然科学基金(81170415)

摘　　要：目的研究粒细胞巨噬细胞集落刺激因子(granulocyte-macrophage colony stimulating factor,GM-CSF)分泌型肝癌疫苗对移植性肝癌小鼠细胞毒性T淋巴细胞杀伤活性的作用机制。方法本实验建立3个组别:肝癌疫苗组(A组),肝癌组(B组)及PBS对照组(C组)。在小鼠体内注入H 22肝癌细胞构建移植性肝癌组,同时建立GM-CSF分泌型肝癌疫苗组及PBS对照组;采用流式细胞术检测每组小鼠外周血中CD8+T细胞免疫效应分子的表达水平;采用MTT方法检测各组脾细胞细胞毒T淋巴细胞(CTL)的杀伤活性;采用Western blot方法检测肿瘤坏死因子-α(tumor necrosis factor-α,TNF-α)和γ-干扰素(γ-interferon,γ-INF)的表达水平。结果流式细胞术结果表明,与肝癌组相比,GM-CSF分泌型肝癌疫苗可显著地增加小鼠外周血中CD8+T细胞的表达(P<0.01);MTT结果表明,与肝癌组相比,GM-CSF分泌型肝癌疫苗可显著地增加脾细胞细胞毒T淋巴细胞的杀伤活性(P<0.01);Western blot结果表明,与肝癌组相比,GM-CSF分泌型肝癌疫苗组可显著下调TNF-α和γ-INF蛋白的表达(P<0.01)。结论 GM-CSF分泌型肝癌细胞疫苗可显著地抑制H22肝癌细胞H22的活性,其作用机制可能是通过活化CD8+T细胞的表达和提高脾细胞细胞毒T淋巴细胞的杀伤活性,降低TNF-α和γ-INF蛋白的表达。Objective To study the effect of granulocyte-macrophage colony-stimulating factor （ GM-CSF） secreting liver cancer vaccine on killing activity of cytotoxic T lymphocytes （ CTL） of transplanted liver cancer mice and its mechanism.Methods There were three groups：liver cancer vaccine group （A group）, liver cancer group （B group） and PBS group （C group）.The transplanted liver cancer model was builded with injection of H 22 hepatoma cells, while the GM-CSF secreting liver cancer vaccine group and PBS group was builded.GM-CSF secreting liver cancer vaccine group and PBS group were establised.The levels of CD8 ＋T cell in peripheral blood were detected by flow cytometry.The killing activity of cytotoxic T lymphocytes （ CTL） of spleen cells was detected by MTT method.The expression levels of tumor necrosis factor-α（ TNF-α） and interferon-γ（γ-INF） were detected by Western blot.Results The flow cytometry results showed that, compared with B group, the levels of CD8 ＋T cell of A group significantly increased （P〈0.01）.MTT results showed that, compared with B group, the killing activity of cytotoxic T lymphocytes （CTL） in A group significantly increased （P〈0.01）.Western blot results showed that, compared with B group, the expression levels of tumor necrosis factor-α（TNF-α） and interferon-γ（γ-INF） in A group significantly decreased （P〈0.01）.Conclusion GM-CSF secreting liver cancer vaccine can significantly inhibit the activity of H22 cell, and its possible mechanism of action may be to activated CD8 ＋T expression, improve cytotoxic activity of CTL of spleen cells, and reduce TNF-αand γ-INF protein expression.

关 键 词：粒细胞巨噬细胞集落刺激因子 移植型肝癌 分泌型肝癌疫苗 细胞毒性T淋巴细胞 

分 类 号：R735.7[医药卫生—肿瘤]