半夏泻心汤联合IL-12转染骨髓间充质干细胞对胃癌荷瘤裸鼠抑瘤作用研究

Study on Antitumor Effects of Banxia Xiexin Decoction Combined with IL-12 Transfected BMSCs on Nude Mice with Gastric Cancer

目的观察半夏泻心汤联合IL-12转染骨髓间充质干细胞(BMSCs)对胃癌荷瘤裸鼠的抑瘤作用。方法取培养第3代的BMSCs,采用LV5空载慢病毒及LV5 IL-12过表达慢病毒转染BMSCs细胞,分别于转染48、72 h后,倒置荧光显微镜观察,经嘌呤霉素筛选稳定转染BMSCs,RT-qPCR检测BMSCs中IL-12的表达。Balb/C裸鼠腋窝皮下接种人胃癌BCG-823细胞建立胃癌荷瘤裸鼠模型,实验分为成瘤组、空载组、转染组、药物组、联合组。空载组、转染组和联合组经尾静脉注射0.2 mL含1×10^7/mL转染慢病毒的BMSCs悬液,药物组和联合组给予半夏泻心汤27.43 g/kg灌胃,每次0.5 mL,成瘤组给予等量生理盐水灌胃,隔日测量并计算瘤体体积,连续干预13 d,取瘤体组织,计算抑瘤率。结果倒置荧光显微镜下稳定转染病毒的BMSCs呈不规则形、星形,蜂窝状绿色荧光分布,经RT-qPCR验证,转染BMSCs的IL-12 mRNA表达升高,与正常BMSCs、空载BMSCs比较差异有统计学意义(P<0.05);与成瘤组、空载组比较,转染组、药物组及联合组瘤体积及瘤质量增加较小(P<0.05);与转染组、药物组比较,联合组瘤体积及瘤质量增加最小(P<0.05);与空载组比较,转染组、药物组及联合组抑瘤率明显升高(P<0.05);与转染组、药物组比较,联合组抑瘤率明显升高(P<0.05)。结论半夏泻心汤及IL-12转染的BMSCs对胃癌荷瘤裸鼠均有一定的抑瘤作用,二者联合具有协同增效作用。

Objective To observe the antitumor effects of Banxia Xiexin Decoction combined with IL-12 transfected BMSCs on nude mice with gastric cancer.Methods BMSCs of the third generation were cultured and transfected by LV5 no-load lentiviruses and IL-12 overexpression lentiviruses into BMSCs.The expression of IL-12 in BMSCs was detected by RT-qPCR after transfected 48 hours and 72 hours through inverted fluorescence microscopy.A nude mouse model of gastric cancer was established by subcutaneous inoculation of human gastric cancer BCG-823 cells into the axilla of Balb/C nude mice.The model was divided into tumorigenic group,no-load group,transfection BMSCs,medicine group and combination group.No-load BMSCs,transfection group and combination group were injected 0.2 mL BMSCs suspension containing 1×10^7/mL lentivirus via tail vein respectively.The medicine group and combination group were given Banxia Xiexin Decoction for gavage(27.43 g/kg,0.5 mL each time).The tumorigenic group was given the same dose of saline for gavage.Tumor volume of each group was measured and calculated every other day.After 13 days of continuous intervention,tumor tissue was taken to calculate the antitumor rate.Results BMSCs stably transfected with the virus showed irregular,star-shaped and honeycomb-like green fluorescence distribution under inverted fluorescence microscopy.The expression of IL-12 mRNA in the transfection BMSCs was increased by RT-qPCR,and there was statistical significance between the normal BMSCs and no-load BMSCs(P<0.05).Compared with tumorigenic group and no-load group,the tumor mass and tumor weight of the transfection group,the medicine group and the combination group increased little(P<0.05);Compared with the transfection group and the medicine group,the tumor mass and tumor weight of the combination group were the least(P<0.05);Compared with the no-load group,the antitumor rates of the transfection group,the medicine group and the combination group significantly increased(P<0.05).Compared with the transfection group and the medicine group,the antitumor rate of the combination group significantly increased(P<0.05).Conclusion Banxia Xiexin Decoction combined with BMSCs transfected with IL-12 have certain anti-tumor effects on nude mice with gastric cancer,and their combination has a significant synergistic effect.