[1]郭　琪,胡　娇,张　毅,等.5′-氮杂-2′-脱氧胞苷对人肺鳞状细胞癌细胞株NCI-H226和NCI-H520中CNRIP1基因甲基化及蛋白表达的影响[J].新乡医学院学报,2020,37(5):401-403.[doi:10.7683/xxyxyxb.2020.05.001]
　GUO Qi,HU Jiao,ZHANG Yi,et al.Effect of 5′-aza-2′-deoxycytidine on methylation status and protein expression of cannabinoid receptor interacting protein 1 gene in human lung squamous carcinoma cell lines NCI-H226 and NCI-H520[J].Journal of Xinxiang Medical University,2020,37(5):401-403.[doi:10.7683/xxyxyxb.2020.05.001]
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5′-氮杂-2′-脱氧胞苷对人肺鳞状细胞癌细胞株NCI-H226和NCI-H520中CNRIP1基因甲基化及蛋白表达的影响
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《新乡医学院学报》[ISSN:1004-7239/CN:41-1186/R]

卷:
37
期数:
2020年5
页码:
401-403
栏目:
基础研究
出版日期:
2020-05-05

文章信息/Info

Title:
Effect of 5′-aza-2′-deoxycytidine on methylation status and protein expression of cannabinoid receptor interacting protein 1 gene in human lung squamous carcinoma cell lines NCI-H226 and NCI-H520
作者:
郭　琪1; 胡　娇2; 张　毅1; 胡明军1
（1.西安市胸科医院外科，陕西　西安　710000；2.西安市胸科医院中医康复科，陕西　西安　710000）
Author(s):
GUO Qi1; HU Jiao2; ZHANG Yi1; HU Mingjun1
（1.Department of Surgery，Xi′an Chest Hospital，Xi′an 710000，Shaanxi Province,China；2.Department of Rehabilitation of Traditional Chinese Medicine，Xi′an Chest Hospital,Xi′an 710000,Shaanxi Province,China）
关键词:
NCI-H226; NCI-H520; 人肺鳞状细胞癌; 基因甲基化; 大麻素受体相互作用蛋白-1; 5′-氮杂-2′-脱氧胞苷
Keywords:
NCI-H226; NCI-H520; human lung squamous carcinoma; gene methylation; cannabinoid receptor interacting protein 1;  5′-aza-2′-deoxycytidine
分类号:
R734.2
DOI:
10.7683/xxyxyxb.2020.05.001
文献标志码:
A
摘要:
目的　探讨5′-氮杂-2′-脱氧胞苷对人肺鳞状细胞癌细胞株NCI-H226和NCI-H520中大麻素受体相互作用蛋白-1（CNRIP1）基因甲基化和蛋白表达的影响。方法　取对数生长期NCI-H226和NCI-H520细胞株分为实验组和对照组，对照组细胞使用不含5′-氮杂-2′-脱氧胞苷的培养基继续培养；将实验组细胞分为0.5、1.0、1.5 μmol·L-1组，分别使用含不同浓度（0.5、1.0、1.5 μmol·L-1）5′-氮杂-2′-脱氧胞苷的培养基继续培养。取各组培养72 h后的细胞进行DNA提取及蛋白提取，采用甲基化特异性聚合酶链反应法检测各组细胞中CNRIP1基因的甲基化状态， 采用Western blot检测各组细胞中CNRIP1蛋白表达。结果　对照组和0.5 μmol·L-1组NCI-H226、NCI-H520细胞株均显示为甲基化， 1.0、1.5 μmol·L-1组中2种细胞株均显示为非甲基化。0.5、1.0、1.5 μmol·L-1组2种细胞株中CNRIP1蛋白表达量均高于对照组（P<0.05）；0.5、1.0、1.5 μmol·L-1组2种细胞株中CNRIP1蛋白表达量随浓度的增加而升高，两两比较差异均有统计学意（P<0.05）。结论　人肺鳞状细胞癌细胞株NCI-H226和NCI-H520中CNRIP1基因发生甲基化可能是导致基因表达下调甚至失活的主要原因，5′-氮杂-2′-脱氧胞苷可以逆转CNRIP1基因甲基化状态，促进CNRIPI蛋白表达。
Abstract:
Objective　To investigate the methylation status of cannabinoid receptor interacting protein 1（CNRIP1）gene in human lung squamous carcinoma cell lines NCI-H226，NCI-H520 and the effect of 5′-aza-2′-deoxycytidine on the methylation status and protein expression of CNRIP1.Methods　The NCI-H226 and NCI-H520 cell lines in logarithmic growth period were divided into experimental group and control group.The cells in control group were cultured in the medium without 5 ′- aza-2′ - deoxycytidine.The cells in experimental group were divided into 0.5,1.0,1.5 μmol·L-1 group which were cultured in the medium with different concentrations (0.5,1.0,1.5 μmol·L-1) of 5 ′- aza-2′ - deoxycytidine.After 72 hours of culture,the cells were taken for DNA extraction and protein extraction.Then the methylation status of CNRIP1 gene in each group was detected by methylation specific polymerase chain reaction；the expression of CNRIP1 protein in the cells in each group was detected by Western blot.Results　The cell lines of NCI-H226 and NCI-H520 in control group and 0.5 μmol·L-1 group were methylation,while in the 1.0,1.5 μmol·L-1 group were non-methylation.The expressions of CNRIP1 protein in NCI-H226 and NCI-H520 cell lines in 0.5,1.0,1.5 μmol·L-1 group were higher than those in the control group (P<0.05);the expression of CNRIP1 protein in NCI-H226 and NCI-H520 cell lines in 0.5,1.0,1.5 μmol·L-1 group increased with the increase of concentration,and there was statistic difference among the three groups (P<0.05).Conclusion　The methylation of CNRIP1 gene in human lung squamous carcinoma cell lines NCI-H226 and NCI-H520 may be the main reason of the down-regulation or even inactivation of gene expression.The 5′-aza-2′-deoxycytidine can reverse the methylation of CNRIP1,and promote the expression of CNRIP1 protein.

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更新日期/Last Update: 2020-05-05