右旋美托咪定抑制胰岛素样生长因子2通路对卵巢癌大鼠免疫功能和癌细胞侵袭迁移的影响及调控机制研究

Effects and regulation mechanism research of dexmedetomidine on insulin-like growth factor 2 pathway on immune function and cancer cell invasion and migration in ovarian cancer rats

目的:探究右旋美托咪定抑制胰岛素样生长因子2(IGF2)受体通路对卵巢癌大鼠免疫功能和癌细胞侵袭迁移的影响机制。方法:选取雌性Fischer 344实验大鼠,正常大鼠作为对照组,参与实验的大鼠于皮下接种NUTU-19低分化上皮性卵巢癌建立卵巢癌大鼠模型,并将成功建模的大鼠随机分为模型组及右旋美托咪定治疗组,右旋美托咪治疗组腹膜内注射右旋美托咪定,1次/d,持续15 d,对照组和模型组给予等体积生理盐水。术后采用RT-q PCR实时分析卵巢组织IGF2、IGF1R和IRS1 m RNA表达,Western blot检测38MAPK/NF-κB信号通路蛋白表达,流式细胞仪检测CD4和CD8 T细胞亚群含量,ELISA检测血清IL-2和TNF-α水平,Transwell试验和刮擦黏附试验检测卵巢癌细胞侵袭和迁移能力,MTT检测右旋美托咪定对脾淋巴细胞增殖的影响。结果:与模型组相比,右旋美托咪定治疗组IGF2、IGF1R和IRS1 m RNA表达降低(P<0.05)。模型组较对照组和右旋美托咪定治疗组p38、NF-κB蛋白表达升高(P<0.05)。右旋美托咪定治疗组CD4和CD8 T细胞亚群百分比较模型组升高,且模型组CD4^(+)与CD8^(+)细胞比值降低程度明显小于右旋美托咪定组(P<0.05)。右旋美托咪定治疗组较模型组血清IL-2和TNF-α水平降低(P<0.05)。右旋美托咪定治疗组较模型组细胞迁移率、侵袭率降低。模型组较右旋美托咪定治疗组脾淋巴细胞增值率降低(P<0.05)。结论:右旋美托咪定可有效抑制IGF2信号通路活化,改善卵巢癌大鼠免疫功能,还可通过降低p38、NF-κB蛋白表达和血清IL-2和TNF-α水平抑制卵巢癌细胞侵袭和迁移。

Objective:To explore mechanism of dexmedetomidine inhibiting insulin-like growth factor 2(IGF2)receptor pathway on immune function and cancer cell invasion and migration in ovarian cancer rats.Methods:Female Fischer 344 experimental rats were selected.Normal rats were used as control group,experimental rats were subcutaneously inoculated with NUTU-19 poorly differentiated epithelial ovarian cancer to establish rat model of ovarian cancer.Successfully modeled rats were randomly divided into model group and dexmedetomidine treatment group.Dexmedetomidine treatment group was intraperitoneal injected of dexmedetomidine once a day for 15 days,control group and model group were given same volume of saline.RT-qPCR was used to analyze expressions of IGF2,IGF1 R and IRS1 mRNA in ovarian tissues in real time after operation.Expressions of p38 MAPK/NF-κB signaling pathway proteins were determined by Western blot.CD4 and CD8 T cells contents were detected by flow cytometry,serum IL-2 and TNF-αlevels were measured by ELISA,ovarian cancer cell invasion and migration ability were measured by Transwell test and scratch adhesion test,and effect of dexmedetomidine on spleen of lymphocyte proliferation was detected by MTT.Results:Compared with model group,expressions of IGF2,IGF1 R and IRS1 mRNA in dexmedetomidine treatment group were decreased(P<0.05).Expressions of p38 and NF-κB proteins in model group were higher than that in control group and dexmedetomidine treatment group(P<0.05).Percentages of CD4 and CD8 T cell subsets in dexmedetomidine treatment group were higher than those in model group,and degrees of reduction in ratios of CD4^(+)to CD8^(+)in model group were significantly smaller than that in dexmedetomidine group(P<0.05).Serum IL-2 and TNF-αlevels were lower in dexmedetomidine treatment group than in model group(P<0.05).Dexmedetomidine treatment group had lower cell migration rate and cell invasion rate than model group.Spleen lymphocyte proliferation rate of model group was lower than that of dexmedetomidine treatment group(P<0.05).Conclusion:Dexmedetomidine can effectively inhibit activation of IGF2 signaling pathway and improve immune function of ovarian cancer rats,and inhibit invasion and migration of ovarian cancer cells by reducing expressions of p38,NF-κB protein and serum IL-2 and TNF-α.