5-氮杂-2′-脱氧胞苷对人甲状腺癌裸鼠移植瘤的抑制作用及其机制的研究  

The inhibitory effect of 5-aza-2'-deoxycytidine on xenograft tumor of human thyroid carcinoma in nude mice and its mechanism

作　　者：程冉 魏枫[1] 张苑 刘美岚[1] 贺佳 CHENG Ran;WEI Feng;ZHANG Yuan;LIU Meilan;HE Jia(Department of Endocrinology,the First Affiliated Hospital of Baotou Medical College,Inner Mongolia University of Science&Technology,Baotou 010400,China)

机构地区：[1]内蒙古科技大学包头医学院第一附属医院内分泌科,014000

出　　处：《天津医药》2022年第5期476-481,共6页Tianjin Medical Journal

基　　金：内蒙古自治区科技计划项目(201802128)。

摘　　要：目的观察去甲基化制剂5-氮杂-2′-脱氧胞苷(5-Aza-CdR)对人甲状腺乳头状癌TPC-1细胞裸鼠移植瘤的生长抑制作用。方法将人甲状腺乳头状癌TPC-1细胞接种于16只BALB/c雌性裸鼠右前肢腋下。移植瘤模型建立后,用随机数字表法将裸鼠分为2组,实验组按照1μg/g腹腔注射5-Aza-CdR 200μL,每2 d给药1次,给药4周。对照组腹腔注射等体积生理盐水。末次给药后处死荷瘤鼠,观察2组裸鼠移植瘤的生长情况,HE染色观察移植瘤的病理形态学改变,采用甲基化特异性PCR检测死亡相关蛋白激酶(DAPK)基因甲基化程度,采用免疫组化染色与Western blot检测DAPK和DNA甲基转移酶(DNMTs)的蛋白表达水平。结果经药物干预4周后,实验组移植瘤体积和质量均较对照组明显减少(P<0.05),相比对照组,实验组抑瘤率为45.38%。HE染色结果显示,实验组肿瘤细胞数量减少,部分区域肿瘤细胞核淡染、缩小,局部出现核固缩、核溶解。实验组DAPK基因甲基化程度低于对照组。免疫组化染色和Western blot结果显示,实验组DAPK蛋白表达量明显高于对照组,DNMT1、DNMT3A蛋白表达量低于对照组,差异有统计学意义(均P<0.05),2组DNMT3B的表达差异无统计学意义(P>0.05)。结论5-Aza-CdR可通过调节裸鼠移植瘤内DNMTs来提高DAPK的表达,从而抑制甲状腺乳头状癌生长。Objective To observe the inhibitory effect of 5-aza-2'-deoxycytidine(5-Aza-CdR)on the growth of human papillary thyroid carcinoma TPC-1 cells in nude mice.Methods Human papillary thyroid carcinoma TPC-1 cells were inoculated subcutaneously under the armpit of right forelimb in 16 female BALB/C nude mice.After the model was established,the nude mice were divided into 2 groups.The experimental group was intraperitoneally injected with 5-Aza-CdR 200μL per 1μg/g,and the control group was intraperitoneally injected with iso-volume sterile normal saline,once every 2 days for 4 weeks.The tumor-bearing mice were killed after the last drug intervention.The growth of transplanted tumors in nude mice of the two groups was observed.The histopathological changes of transplanted tumors were observed by HE staining.Methylation-specific PCR was used to detect the methylation degree of death associated protein kinase(DAPK)gene,and the protein expression levels of DAPK and DNA methyltransferases(DNMTs)were detected by immunohistochemical staining and Western blot assay.Results After 4 weeks of drug intervention,the tumor volume and weight were significantly reduced in the experimental group compared with the control group(P<0.05),and the tumor inhibition rate was 45.38%in the experimental group compared with the control group.HE staining results showed that the number of tumor cells in the experimental group was reduced,the nuclei of tumor in some areas were faint and shrunken,and nuclear pyknosis and nucleolysis occurred locally.DAPK gene methylation was lower in the experimental group than that in the control group.Immunohistochemical staining and Western blot results showed that the expression of DAPK protein was significantly higher in the experimental group than that in the control group,while the expression levels of DNMT1 and DNMT3A protein were significantly lower than those in the control group(P<0.05).There was no significant difference in the expression of DNMT3B between the two groups(P>0.05).Conclusion 5-Aza-CdR can

关 键 词：甲状腺肿瘤 癌 乳头状 死亡相关蛋白激酶类 DNA甲基化 小鼠 近交BALB C 5-氮杂-2′-脱氧胞苷 移植瘤 DNA甲基转移酶 

分 类 号：R736.1[医药卫生—肿瘤]