丙氨瑞林对人子宫内膜癌裸鼠皮下移植瘤生长的影响及对Smurf2表达的影响

发布时间：2018-06-09 21:16

本文选题：子宫内膜癌 + 丙氨瑞林　；参考：《山西医科大学》2014年硕士论文

【摘要】：研究目的通过构建人子宫内膜癌裸鼠皮下移植瘤动物模型，探讨促性腺激素释放激素（GnRH）类似物——醋酸丙氨瑞林（Alarelin Acetate）对人子宫内膜癌裸鼠移植瘤的生长及对Smurf2表达的影响。使用3种不同剂量的丙氨瑞林对人子宫内膜癌裸鼠移植瘤模型进行分组干预实验，并探讨药物抑制对Smurf2表达调控的影响。研究方法将体外培养冻存的人子宫内膜癌Hec-1B细胞复苏置于适当的环境中培养，当细胞培养到一定比例时接种于4-6周龄雌性裸鼠左前肢近腋窝处皮下，构建形成皮下移植瘤，成功建模后将24只裸鼠随机分组，共4组，每组6只：第1组为对照组，肌肉注射生理盐水，第2组为丙氨瑞林低剂量组(14g/kg)，第3组为中剂量组(28g/kg)，第4组为高剂量组(56g/kg)。丙氨瑞林给药方式为背部肌肉注射，，每只裸鼠根据体重计算所需用药剂量，调整注射液体量为每只裸鼠注射0.1ml，每日给药1次，连续给药4周。在此期间4天测量1次裸鼠瘤体的最长径与最短径，计算瘤体体积，绘制肿瘤生长曲线、同时观察裸鼠整体生长状况。治疗4周后处死裸鼠，取出移植瘤，计算不同剂量丙氨瑞林组的抑瘤率。将移植瘤组织制成石蜡切片，进行HE染色，在光学显微镜下观察肿瘤组织形态学情况。用免疫组织化学方法检测皮下移植瘤组织中Smurf2蛋白的表达情况。实验结果 1.对照组、丙氨瑞林低、中、高剂量组的裸鼠皮下移植瘤生长曲线斜率依次减小，生长速度依次减慢，差异有统计学意义(P0.05)； 2.药物干预4周后，移植瘤体积分别是：对照组1628±162mm3，低剂量组1368±256mm3、中剂量组1296±307mm3、高剂量组995±213mm3，差异有统计学意义(P0.05)。丙氨瑞林低、中、高剂量组的抑瘤率分别15.94%、20.37%、38.86%，组间两两比较，差异有统计学意义(P0.05)； 3.移植瘤组织HE染色结果：光学显微镜下观察可知，对照组移植瘤组织结构排列紊乱，细胞体积较小，有明显异型性，且间质少，细胞核体积和核浆比也明显增大，分布不均，染色较深，可见较多核分裂像；治疗组移植瘤细胞排列较稀疏，可见多处无结构的红染区，瘤细胞核深染、固缩。 4.免疫组织化学结果：丙氨瑞林可增加Smurf2蛋白在移植瘤组织中表达水平，并且随着丙氨瑞林剂量的增加，Smurf2的表达逐渐增强（P0.05），且组间两两比较差异有统计学意义（P0.05）。实验结论 1.丙氨瑞林能够有效抑制人子宫内膜癌细胞株裸鼠皮下移植瘤的生长，且与剂量效应呈正相关。 2.丙氨瑞林对人子宫内膜癌裸鼠移植瘤的抑制作用可能是通过上调Smurf2的表达来实现的。
[Abstract]:Objective to investigate the effects of gonadotropin releasing hormone (GnRH) analogue-Alarelin Acetateon the growth of human endometrial carcinoma in nude mice and the expression of Smurf2 in nude mice. Three different doses of alarelin were used to treat human endometrial carcinoma xenografts in nude mice. To investigate the effect of drug inhibition on the expression and regulation of Smurf2.Methods the cryopreserved human endometrial carcinoma Hec-1B cells were resuscitated in a suitable environment. When the cells were cultured to a certain proportion, they were inoculated subcutaneously in the left forelimb near axilla of 4-6 weeks old female nude mice. After successful modeling, 24 nude mice were randomly divided into 4 groups, 6 in each group: the first group was the control group. Normal saline was injected intramuscularly in the second group, in the low dose group (14 g / kg), in the third group in the middle dose group (28 g / kg), in the fourth group in the high dose group (56 g / kg). Alarelin was injected intramuscularly on the back of each nude mouse according to its body weight. The dosage of the injection was adjusted to 0.1 ml per nude mouse once a day for 4 weeks. The longest and shortest diameters of nude mice tumor were measured once in 4 days, the tumor volume was calculated, the tumor growth curve was drawn, and the whole growth state of nude mice was observed at the same time. After 4 weeks of treatment, nude mice were killed, transplanted tumor was removed and tumor inhibition rate was calculated in different doses of alarelin group. The transplanted tumor tissue was made into paraffin sections and stained with HE, and the morphology of the tumor was observed under optical microscope. The expression of Smurf2 protein in subcutaneous transplanted tumor was detected by immunohistochemical method. In the control group, the slope of growth curve of subcutaneous transplanted tumor decreased in turn, and the growth rate of subcutaneous transplanted tumor in the control group decreased in turn, and the difference was statistically significant (P 0.05). After 4 weeks of drug intervention, the volume of transplanted tumor was 1628 卤162mm3 in control group, 1368 卤256mm3 in low dose group, 1296 卤307mm3 in middle dose group and 995 卤213mm3 in high dose group. The difference was statistically significant (P 0.05). The tumor inhibition rate of the low, medium and high dose groups was 15.9420.37 and 38.86, respectively. The difference between the two groups was statistically significant (P 0.05; 3. 0. 5%, P < 0. 05; P < 0. 05; P < 0. 05; P < 0. 05; P < 0. 05). The results of HE staining showed that the structure of the transplanted tumor in the control group was disordered, the size of cells was small, there was obvious heterogeneity, and the interstitial size was less, the nuclear volume and the ratio of nucleus to cytoplasm were also increased obviously, and the distribution was uneven. In the treatment group, the transplanted tumor cells were arranged sparsely, and there were many unstructured red staining areas. The nuclei of the treatment group were deep stained and pyknotic. Immunohistochemical results: alarelin could increase the expression of Smurf2 protein in transplanted tumor tissues, and the expression of Smurf2 increased gradually with the increase of the dose of alarelin, and the difference between the two groups was statistically significant (P 0.05). Conclusion 1. Alarelin could effectively inhibit the growth of human endometrial carcinoma cell line in nude mice, and had a positive correlation with dose effect. 2. The inhibitory effect of alarelin on human endometrial carcinoma xenografts in nude mice may be achieved by upregulating the expression of Smurf2.
【学位授予单位】：山西医科大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R737.33

【相似文献】