冷冻消融联合索拉非尼和IL-2治疗裸鼠肾癌移植瘤的实验研究

Experimental study on the therapeutic effects of cryoablation combined with sorafenib and interleukin 2 on renal cancer transplantation tumor in nude mice

目的观察冷冻消融联合索拉非尼和IL-2治疗裸鼠肾癌移植瘤的效果及可能作用机制。方法建立人肾癌Renca裸鼠皮下移植瘤模型,共50只,随机分为对照组、冷冻消融组、索拉非尼组、IL-2组、联合治疗组,每组10只。接种4周后处死裸鼠,剥离肿瘤组织,比较5组肿瘤组织瘤重,计算抑瘤率。流式细胞术检测5组肿瘤细胞凋亡率及T淋巴细胞亚群、NK细胞比例。ELISA法检测5组血清VEGF、VEGFR-2、HIF-1α水平。Western blot检测5组肿瘤组织血管内皮生长因子(VEGF)、VEGFR-2、HIF-1α蛋白表达。免疫组织化学SP法检测5组肿瘤组织微血管密度(MVD)。结果联合治疗组肿瘤重量、血清VEGF、VEGFR-2、HIF-1α、水平、癌组织VEGF、VEGFR-2、HIF-1α蛋白表达及MVD低于其他4组,抑瘤率、肿瘤细胞凋亡率、外周血CD+4、CD+8T淋巴细胞亚群、NK细胞比例高于其他4组,生存时间长于其他4组,差异有统计学意义(P<0.05)。结论冷冻消融联合索拉非尼和IL-2治疗能够通过抗血管生成、增强免疫功能抑制裸鼠肾癌移植瘤生长。

Objective To investigate the therapeutic effects of cryoablation combined with sorafenib and interleukin 2 on renal cancer transplantation tumor in nude mice,and to explore possible action mechanism. Methods The subcutaneous transplantation tumor models of human renal cancer（ Renca） were established in 50 nude mice. The nude mice were randomly divided into control group,cryoablation group,sorafenib group,interleukin 2 group and combination treatment group,with 10 mice in each group. At four weeks after inoculating,these nude mice were sacrificed and the tumor tissues were separated.The weight of tumor tissues and inhibition rates of tumor growth were calculated and compared. The apoptosis rates of tumor cells,the percentages of T lymphocyte subpopulation and natural killer（ NK） cells proportion were detected by flow cytometry. The serum levels of vascular endothelial growth factor（ VEGF）,VEGFR 2,hypoxia-inducible factor 1α（ HIF 1α）were measured by enzyme-linked immunosorbent assay（ ELISA）. The expression levels of VEGF,VEGFR 2 and HIF 1αproteins in tumor tissues were detected by Western Blot. The microvessel density（ MVD） of tumor tissues was detected by immunohistochemical SP method. Results The tumor weigh,serum levels of VEGF,VEGFR 2,HIF 1α,expression levels of VEGF,VEGFR 2,HIF 1α proteins and MVD of tumor tissues in combination treatment group were significantly lower than those in the other groups（ P〈0. 05）. However the inhibition rates of tumor growth,apoptosis rates of tumor cells and the percentages of CD_4~＋T,CD＋8~＋ T and NK cells in combination treatment group were significantly higher than those in the other groups（ P〈0. 05）. Moreover the survival time of mice in combined treatment group was significantly longer than that in the other groups（ P〈0. 05）. Conclusion The cryoablation combined with sorafenib and interleukin 2 can inhibit the growth of renal cancer transplantation tumor in nude mice through anti-angiogenesis and immune-enhancement.