阿霉素联合二甲双胍对人舌癌裸鼠移植瘤的抑制作用及相关机制的研究

发布时间：2018-05-02 06:42

本文选题：舌癌 + 裸鼠移植瘤　；参考：《广西医科大学》2017年硕士论文

【摘要】：目的:研究阿霉素(Adriamycin,ADM)联合二甲双胍(Metformin,MET)对人舌癌裸鼠移植瘤的抑制作用及对Ki67、Cyclin D1、VEGF分子表达的影响,并探讨其相关的抗肿瘤作用机制。方法:建立人舌癌SCC-25细胞裸鼠移植瘤模型,将24只裸鼠随机分为对照组、二甲双胍组、阿霉素组、阿霉素联合二甲双胍组,按照给药方案用药,观察各组裸鼠的一般生活状态,每隔3天记录其体重变化及游标卡尺测量移植瘤的体积,绘制生长曲线。给药15天后处死裸鼠,测量肿瘤体积并计算各组抑瘤率,实时荧光定量PCR检测各组移植瘤Cyclin D1、VEGF m RNA的相对表达,免疫组化法检测各组肿瘤组织Ki67抗原的表达情况。结果:1.用药结束后,阿霉素组及联合用药组平均体重明显低于对照组及二甲双胍组,差异有统计学意义(P0.05);阿霉素组及联合用药组平均体重较用药前明显降低,差异有统计学意义(P0.01)。2.用药结束后,二甲双胍组、阿霉素组、联合用药组平均体积分别为(1022.16±262.49)mm3、(666.31±92.13)mm3、(595.06±193.54)mm3、(374.87±129.52)mm33.实时荧光定量PCR检测结果显示:与对照组相比,二甲双胍组、阿霉素组、二甲双胍/阿霉素联合用药组VEGF、Cyclin D1 m RNA表达水平降低,差异有统计学意义(P0.01、P0.05)。与二甲双胍组、阿霉素组比较,,抑瘤率分别为34.81%、41.78%、63.32%,与对照组相比差异有统计学意义(P0.05);联合用药组平均体积小于阿霉素组及二甲双胍组,差异有统计学意义(P0.05)。二甲双胍/阿霉素联合用药组VEGF、Cyclin D1 m RNA表达水平降低,差异有统计学意义(P0.05)。4.免疫组化结果显示:与对照组相比,二甲双胍组、阿霉素组、阿霉素联合二甲双胍组Ki67抗原阳性表达不同程度降低,差异有统计学意义(P0.01)。二甲双胍/阿霉素联合用药组Ki67抗原阳性表达率最低,与二甲双胍、阿霉素组相比差异有统计学意义(P0.01)。结论:1.阿霉素联合二甲双胍较阿霉素单用药对人舌癌SCC-25细胞移植瘤有更显著的抗肿瘤作用。2.二甲双胍及阿霉素均能抑制人舌癌SCC-25移植瘤的生长,推测可能是通过降低移植瘤中VEGF、Cyclin D1的表达从而发挥抑制肿瘤血管生成及诱导细胞周期阻滞的抗肿瘤作用,而联合用药对移植瘤中VEGF、Cyclin D1的下调水平更显著,抑瘤效果更好。
[Abstract]:Aim: to study the inhibitory effect of adriamycin (ADM) combined with metformin (MET) on the expression of VEGF in human tongue carcinoma in nude mice and to explore the mechanism of its antitumor effect. Methods: the transplanted tumor model of human tongue cancer SCC-25 cells was established in nude mice. 24 nude mice were randomly divided into control group, metformin group and adriamycin combined with metformin group. Body weight change and Vernier caliper were recorded every 3 days to measure the volume of transplanted tumor and draw the growth curve. After 15 days of administration, nude mice were killed, tumor volume was measured and tumor inhibition rate was calculated. The relative expression of Cyclin D1 PCR was detected by real-time quantitative PCR, and the expression of Ki67 antigen in tumor tissues was detected by immunohistochemical method. The result is 1: 1. After treatment, the average body weight of adriamycin group and combination group was significantly lower than that of control group and metformin group, the difference was statistically significant (P 0.05), and the average body weight of adriamycin group and combination group was significantly lower than that of before treatment. After the treatment, the average volume of metformin group, adriamycin group and combination group were 1022.16 卤262.49 mm ~ (3), 666.31 卤92.13 ~ (13) mm ~ (3), 595.06 卤193.54 mm ~ (3), 374.87 卤129.52 m ~ (33), respectively. The results of real-time fluorescence quantitative PCR showed that the expression of VEGF cyclin D1 m RNA in metformin group, adriamycin group and metformin / adriamycin combination group was significantly lower than that in control group (P 0.01). Compared with the metformin group and the adriamycin group, the tumor inhibition rate was 34.81 and 41.78 and 63.32, respectively, and the average volume in the combination group was smaller than that in the adriamycin group and metformin group (P 0.05). The expression of VEGF cyclin D1 m RNA in metformin / doxorubicin group was significantly lower than that in control group (P 0.05. 4). Compared with the control group, the positive expression of Ki67 antigen in metformin group, adriamycin group and adriamycin plus metformin group was significantly lower than that in control group (P 0.01). The positive rate of Ki67 antigen in metformin / adriamycin group was the lowest, and there was significant difference between metformin group and adriamycin group (P 0.01). Conclusion 1. Adriamycin combined with metformin has a more significant antitumor effect than adriamycin alone on human tongue cancer SCC-25 cells. Both metformin and adriamycin can inhibit the growth of human tongue cancer SCC-25 xenografts. It is assumed that both metformin and adriamycin can inhibit angiogenesis and induce cell cycle arrest by reducing the expression of VEGF cyclin D1 in the transplanted tumor. In contrast, the combined use of drugs could significantly down-regulate the level of VEGF cyclin D1 in transplanted tumors, and the inhibitory effect was better.
【学位授予单位】：广西医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R739.86

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