反义STAT3寡核苷酸对B16细胞辐射敏感性的研究

Study on radiosensitivity of B16 cells transfected by STAT3 antisense oligodeoxynucleotides

目的探讨反义STAT3转染鼠恶性黑色素瘤B16细胞后,肿瘤细胞辐射敏感性的变化。方法利用反义寡核苷酸转染B16细胞后,以不同剂量γ射线照射。通过CCK-8试剂盒检测细胞增殖变化,Hoechst33258染色对细胞凋亡作形态学上的观察。用Annexin V/PI复染,流式细胞仪检测细胞早期凋亡率的变化。结果反义STAT3转染后加以γ射线照射,B16细胞的增殖相比于两者单独作用组受到明显抑制,细胞凋亡水平也增加。结论反义STAT3寡核苷酸联合γ射线对B16细胞的增殖抑制和诱导凋亡作用明显增强,提高了鼠黑色素瘤B16细胞的辐射敏感性;表明阻断STAT3蛋白表达可能成为一种新的提高肿瘤辐射敏感性的有效手段。

Objective To investigate the influence of STAT3 antisense oligodeoxynucleotides （ASOSTAT3） on radiosensitivity of B16 cells. Methods Oligodeoxynucleotides were tansfected into B16 cells with oligofectamine reagent. Cell survival was measured using CCK-8 assay. Apoptotic assays were performed using Hoechst33258 staining and Annexin V/PI with FACS analysis. Results Cell survival decreased significantly in those groups tansfected with ASO-STAT3 and irradiated with different dose of γ-rays. Compared with those cells treated with irradiation alone or AO-STAT3 transfection with subsequent exposure to radiation, fraction of early apoptosis increased in those cells treated with ASO-STAT3 transfection combined with irradiation. Conclusion These results showed that targeting ASO-STAT3 can enhance the therapeutic effect of γ-ray irradiation on B16 cells, which implied that STAT3 can act as a potential molecular target for tumor therapies, and also a potential molecular target for enhancement of radiosensitivity of radioresistant tumors.