DNMT抑制剂5-AzaDc对结直肠癌肝转移的影响  

Effects of DNA methyltransferase inhibitor 5-AzaDc on hepatic metastasis of colorectal cancer

作　　者：孙绍伟 张焕虎 曹金 宫庆[1] 韩传吉[1] SUN Shaowei;ZHANG Huanhu;CAO Jin;GONG Qing;HAN Chuanji(Department of Gastrointestinal Surgery,Weihai Municipal Hospital,Cheeloo College of Medicine,Shandong University,Weihai 264200,China)

机构地区：[1]山东大学附属威海市立医院胃肠外科,山东威海264200

出　　处：《山东医药》2022年第1期48-51,共4页Shandong Medical Journal

摘　　要：目的探讨DNA甲基转移酶(DNMT)抑制剂5-氮杂-2′-脱氧胞苷(5-AzaDc)对结直肠癌肝转移的影响。方法选择健康雄性BALB/c裸鼠50只,随机分为模型组、对照组以及5-AzaDc低、中、高剂量组,每组10只。模型组和5-AzaDc低、中、高剂量组通过脾脏种植法构建结直肠癌肝转移模型。建模成功后,5-AzaDc低、中、高剂量组分别腹腔注射1.0μmol/L 5-AzaDc 0.3、0.4、0.5 mg/kg,每周1次,连续注射8周。模型组和对照组腹腔注射等量生理盐水。腹腔注射8周后,颈椎脱臼处死,无菌条件下剖腹取肝转移瘤组织,剪碎、消化、再培养后,经流式细胞仪分离,获取肝转移瘤细胞,检测DNMT活性、肿瘤球数量和细胞存活率;颈椎脱臼处死前,采集外周静脉血,采用ELISA法检测血清CD44v6、VEGF。结果对照组未构建结直肠癌肝转移模型,无肝转移瘤细胞形成。模型组与5-AzaDc低、中、高剂量组肝转移瘤细胞DNMT活性、肿瘤球数量和细胞存活率均逐渐降低(P均<0.05)。5-AzaDc低、中、高剂量组肝转移瘤细胞DNMT活性、肿瘤球数量和细胞存活率均低于模型组(P均<0.05),并且5-AzaDc中、高剂量组低于5-AzaDc低剂量组(P均<0.05),而5-AzaDc中剂量组与5-AzaDc高剂量组比较P均>0.05。模型组和5-AzaDc低、中、高剂量组血清CD44v6、VEGF水平均高于对照组(P均<0.05);5-AzaDc低、中、高剂量组血清CD44v6、VEGF水平均低于模型组(P均<0.05),并且5-AzaDc中、高剂量组均低于5-AzaDc低剂量组(P均<0.05),而5-AzaDc中剂量组与5-AzaDc高剂量组比较P均>0.05。结论5-AzaDc对结直肠癌肝转移具有一定抑制作用,其作用机制可能与抑制肿瘤血管形成有关。Objective To investigate the effect of DNA methyltransferase(DNMT)inhibitor 5-Aza-2′-deoxycytidine(5-AzaDc)on liver metastasis of colorectal cancer.Methods Fifty BALB/c male nude mice were selected,and the mice were randomly divided into the control group,the model group,low-dose,medium-dose,and high-dose 5-AzaDc groups,with 10 in each group.Colorectal cancer mouse models with liver metastases were established by the spleen planting method in the model group,low-dose,medium-dose,and high-dose 5-AzaDc groups.After successful modeling,mice of the low-dose,medium-dose,and high-dose 5-AzaDc groups were intraperitoneally injected with 5-AzaDc(1.0μmol/L)once a week,for 8 weeks,and the doses were 0.3,0.4 and 0.5 mg/kg,respectively;while the mice in the control group and model group were given an equivalent volume injection of physiological saline.At 8 weeks after the intraperitoneal injection,the mice were sacrificed by cervical dislocation,and the tissues of liver metastases were taken by laparotomy under sterile conditions.After the mice were minced,digested and cultured,tumour cells of liver metastases were isolated and obtained by flow cytometry,then the activity of DNMT,the number of tumorspheres and the cell survival rates were detected.Peripheral venous blood samples were collected for each subject before the mice were sacrificed by cervical dislocation,and then the serum CD44v6 and VEGF levels were evaluated by ELISA.Results In the control group,the colorectal cancer mouse model with liver metastases was not constructed,and no tumour cells of liver metastases were formed.DNMT activity of liver metastasis cells,the number of tumorspheres and the cell survival rates all gradually decreased in the model group,low-dose,medium-dose,and high-dose 5-AzaDc groups(all P<0.05).DNMT activity of liver metastasis cells,the number of tumorspheres and the cell survival rates were lower in the low-dose,medium-dose,and high-dose 5-AzaDc groups than those in the model group(all P<0.05),and meanwhile,they were significantly low

关 键 词：结直肠癌 肝转移 DNA甲基转移酶 5-氮杂-2′-脱氧胞苷 小鼠 

分 类 号：R735.3[医药卫生—肿瘤]