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目的 :探讨肿瘤坏死因子受体 (TNFR)基因转移对肿瘤细胞表面 TNFR表达量的影响 ,进而观察其对TNF杀伤肿瘤细胞作用的影响。 方法 :建立 TNFR的逆转录病毒表达载体 ,通过转染包装细胞 PA 317产生完整病毒颗粒 ;人前列腺癌细胞株 PC- 3m和小鼠 Renca肾腺癌细胞株经病毒感染后 ,检测其细胞表面 TNFR数量及TNF在体内外对其杀伤作用的变化。 结果 :TNFR基因转移前后 PC- 3m细胞表面与 TNF结合的位点数量分别为 2 912个 /细胞和 8872个 /细胞 (P<0 .0 5 ) ,Renca细胞表面与 TNF结合的位点数分别为 783个 /细胞和 36 78个 /细胞 (P<0 .0 5 )。 TNFR基因转染后 ,TNF在裸鼠体内对 PC- 3m和 Renca的肿瘤抑制率分别增强 1.8和 2 .1倍。 结论 :以逆转录病毒为载体的基因转移方法 ,可以提高肿瘤细胞表面 TNFR的表达量 ,从而增强 TNF对肿瘤细胞的体外杀伤作用和体内抑制作用
Objective: To investigate the effect of tumor necrosis factor receptor (TNFR) gene transfer on the expression of tumor necrosis factor receptor (TNFR) on the surface of tumor cells, and to observe its effect on tumor cell killing by TNF. METHODS: A retroviral expression vector of TNFR was established, and whole virus particles were generated by transfecting PA 317 into packaging cells; TNFR was detected on human prostate cancer cell line PC-3m and Renca renal adenocarcinoma cell line after infection with virus. Quantity and changes of TNF killing effect in vivo and in vitro. RESULTS: The number of TNF-binding sites on the surface of PC-3m cells before and after TNFR gene transfer was 2,912 cells/cell and 8,872 cells/cell (P<0.05), and the number of sites of Renca cell binding to TNF was 783 cells/cell and 36 78 cells/cell (P<0.05). After transfection of TNFR gene, tumor inhibition rates of PC-3m and Renca by TNF in nude mice were increased by 1.8 and 2.1 times, respectively. Conclusion : The gene transfer method using retrovirus as the vector can increase the expression of TNFR on the surface of tumor cells, thereby enhancing the in vitro killing effect and in vivo inhibitory effect of TNF on tumor cells.