丙氨瑞林对人子宫内膜癌裸鼠移植瘤的作用以及PinX1表达的影响

发布时间：2018-04-17 07:09

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

【摘要】：研究目的促性腺激素释放激素类似物已应用于临床治疗多种肿瘤,但其具体作用机制仍有待于研究。本课题通过观察不同剂量的丙氨瑞林(Alarelin)对子宫内膜癌裸鼠移植瘤的生长抑制和PinX1表达的影响以及两者之间的相互关系,探讨促性腺激素释放激素类似物影响端粒酶促肿瘤通路抑制肿瘤生长可能的作用机制。研究方法人子宫内膜癌细胞株HEC-1B体外培养后配成悬液注射于裸鼠皮下,建立人子宫内膜癌动物模型。26只裸鼠均造模成功,为提高准确性,去掉瘤体结节最大及最小的两只,其余实验荷瘤裸鼠随机地分为四组,标记为低、中、高剂量治疗组及对照组,对照组肌注生理盐水用于对照,治疗组肌注丙氨瑞林进行实验,分别按20ug/kg、40ug/kg、80ug/kg的剂量计算用药量。一次/日,连续治疗4周。实验期间密切观察一般情况,定期测量移植瘤体积,绘制肿瘤生长曲线,判定丙氨瑞林抑制肿瘤情况；实验结束时处死裸鼠,完整剔出裸鼠皮下移植瘤,称得瘤重,并计算抑瘤率,然后将各组每块瘤组织分为两份,一份用来HE染色,观察移植瘤细胞病理变化；另一份用于免疫组化检测PinX1蛋白的表达情况。实验结果 1、肿瘤开始生长及药物干预初期,裸鼠一般情况好,反应佳。随着肿瘤的逐渐增大,各实验组裸鼠均出现消瘦,精神、食欲、活动能力均下降,其中丙氨瑞林高剂量组在实验后期表现最明显的。 2、经过4周干预后,各干预组移植瘤的体积分别是：低剂量组1367.3±111.56mm3、中剂量组1123±117.4mm3、高剂量组982±97.1mm3,与对照组1622.8±154.4mm3相比,有显著性差异(P0.01),低剂量组的抑瘤率为15.72%、中剂量组的抑瘤率为30.76%、高剂量组的抑瘤率为39.46%,中、高剂量组之间的比较瘤体体积和抑瘤率均无显著差异(p=0.056),其他组间比较差异明显(P0.05)。 3、光镜下见对照组间质减少,细胞增生活跃,形态大小不一,异型性明显,核大而深染,可见核分裂象,核浆比例失调,丙氨瑞林治疗组细胞肿胀,异型性小,可见核皱缩、裂解、凋亡、坏死细胞。并可见大片坏死区。 4、对照组中移植瘤组织中PinX1表达极少或缺如,各干预组表达均增高,与对照组比较差异显著(P0.05),,各干预组中,低剂量组表达最低；高剂量组表达最强,两两相比较,仅中高剂量组之间无显著差异(P0.05)。研究结论： 1、丙氨瑞林能够直接抑制子宫内膜癌裸鼠移植瘤的生长,在一定范围内其抑制作用与丙氨瑞林剂量呈正相关。 2、丙氨瑞林可使移植瘤细胞出现明显的病理改变。 3、丙氨瑞林可能是通过上调肿瘤细胞中PinX1的表达,抑制端粒酶活性,使肿瘤细胞端粒得不到延伸,从而实现抑瘤目的。
[Abstract]:Research purposeGonadotropin releasing hormone analogue has been used in the treatment of many kinds of tumors, but its mechanism remains to be studied.In this study, we investigated the effects of different doses of alarelin on growth inhibition and PinX1 expression in nude mice with endometrial carcinoma.To explore the possible mechanism of the effect of gonadotropin releasing hormone analogues on the inhibition of tumor growth by telomerase-stimulated tumor pathway.Research methodHuman endometrial carcinoma cell line HEC-1B was cultured in vitro and injected into nude mice subcutaneously. The animal model of human endometrial carcinoma. 26 nude mice were established successfully. In order to improve the accuracy, the two largest and smallest tumor nodules were removed.The other experimental nude mice were randomly divided into four groups: low, medium and high dose treatment group and control group. The control group was intramuscularly injected with normal saline for the control group, and the treatment group was intramuscularly injected with alarelin. The dosage of the drug was calculated according to the dosage of 20ugr / kg ~ 40ugr / kg / kg ~ 80ugr / kg respectively.Once a day for 4 weeks.During the experiment, we observe the general situation closely, measure the volume of the transplanted tumor regularly, draw the tumor growth curve, and judge the inhibition of the tumor by alarelin. At the end of the experiment, we killed the nude mice, completely removed the transplanted tumor under the skin of the nude mice, and weighed the tumor.The tumor inhibition rate was calculated, then each tumor tissue was divided into two parts, one was used for HE staining to observe the pathological changes of transplanted tumor cells, the other was used to detect the expression of PinX1 protein by immunohistochemistry.Experimental results1. At the beginning of tumor growth and drug intervention, nude mice were generally in good condition and good response.With the increase of tumor, the weight loss, spirit, appetite and activity of the nude mice in each experimental group were all decreased, especially in the high dose group of alarelin in the late stage of the experiment.2. After 4 weeks of intervention, the volume of transplanted tumor in each intervention group was 1367.3 卤111.56 mm 3 in low dose group, 1123 卤117.4 mm 3 in middle dose group and 982 卤97.1mm 3 in high dose group, compared with 1622.8 卤154.4mm3 in control group.There was significant difference in tumor inhibition rate between low dose group (15.72%), middle dose group (30.76%) and high dose group (39.46%). There was no significant difference in tumor volume and tumor inhibition rate between middle and high dose groups (P 0.056).3. Under the light microscope, the interstitial cells of the control group were decreased, the cells proliferated and active, the size of the cells varied, the heterogeneity was obvious, the nucleus was large and deep stained, the mitotic image was seen, the proportion of nuclear and cytoplasm was disordered, the cells in the treatment group were swollen, the dysplasia was small, and the nucleus shrank.Lytic, apoptotic, necrotic cells.A large area of necrosis can be seen.(4) the expression of PinX1 in transplanted tumor tissues in the control group was very little or absent, and the expression of PinX1 in each intervention group was higher than that in the control group, the difference was significant (P 0.05). The expression of PinX1 was the lowest in the low dose group, the strongest in the high dose group, and the highest in both groups.There was no significant difference between middle and high dose groups (P 0.05).The study concluded that:1. Alarelin could directly inhibit the growth of endometrial carcinoma xenografts in nude mice, and the inhibitory effect was positively correlated with the dose of alamrelin in a certain range.2, alarelin could cause obvious pathological changes in transplanted tumor cells.3. Alarelin may inhibit telomerase activity by upregulating the expression of PinX1 in tumor cells, resulting in no extension of telomere in tumor cells, so as to achieve the purpose of tumor inhibition.
【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】：R737.31

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