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目的:观察转染4-1BBL基因的小鼠胃癌细胞株MFC的体内致瘤性及其对小鼠免疫功能的影响。方法:通过脂质体介导以重组质粒pMKITneo和pMKITneo/4-1BBL转染小鼠胃癌细胞株MFC。用RT-PCR法检测目的基因的表达。计数法绘制体外培养细胞生长曲线。将转染前后肿瘤细胞,分别接种于615小鼠背部皮下观察其致瘤性。流式细胞仪测定荷瘤小鼠外周血NK、CD4+T和CD8+T细胞数,并测定荷瘤小鼠肿瘤结节内细胞凋亡率。结果:MFC/4-1BBL细胞中可检测到4-1BBLmRNA表达。转染前后肿瘤细胞的体外生长速度无明显变化。MFC/4-1BBL细胞在小鼠体内的致瘤性较MFC/pMKITneo细胞和野生型MFC细胞明显降低。接种MFC/4-1BBL细胞的小鼠外周血NK细胞及CD8+T细胞数明显增多;其肿瘤结节内细胞凋亡率亦明显增高。结论:转染4-1BBL基因后并未影响MFC细胞的体外生长,但其在小鼠体内的致瘤性明显降低。MFC/4-1BBL细胞可诱导荷瘤小鼠体内CD8+T和NK细胞增殖,而且能促进肿瘤细胞的凋亡。
Objective: To observe the in vivo tumorigenicity of 4-1BBL-transfected murine gastric cancer cell line MFC and its effect on immune function in mice. METHODS: Mouse gastric cancer cell line MFC was transfected with recombinant plasmids pMKITneo and pMKITneo/4-1BBL by liposome. The expression of the target gene was detected by RT-PCR. The counting method was used to draw the cell growth curve in vitro. The tumor cells before and after transfection were inoculated subcutaneously on the back of 615 mice to observe its tumorigenicity. The number of NK, CD4+T and CD8+ T cells in the peripheral blood of the tumor-bearing mice was measured by flow cytometry, and the apoptosis rate of the tumor nodules was measured in the tumor-bearing mice. RESULTS: 4-1BBL mRNA expression was detected in MFC/4-1BBL cells. There was no significant change in the growth rate of tumor cells before and after transfection. The tumorigenicity of MFC/4-1BBL cells in mice was significantly lower than that of MFC/pMKITneo cells and wild-type MFC cells. The number of peripheral blood NK cells and CD8+ T cells in mice inoculated with MFC/4-1BBL cells was significantly increased, and the rate of cell apoptosis in tumor nodules was also significantly increased. Conclusion: The transfection of 4-1BBL gene did not affect the growth of MFC cells in vitro, but its tumorigenicity was significantly reduced in mice. MFC/4-1BBL cells can induce the proliferation of CD8+T and NK cells in tumor-bearing mice, and can promote the apoptosis of tumor cells.