摘要
目的观察甲基化抑制剂5-氮杂-2′-脱氧胞苷(5-aza-2′-deoxycytidine,5-Aza-CdR)对肺癌SPC-A1细胞增殖、细胞划痕和凋亡的影响,探讨抑癌基因分泌型卷曲相关蛋白1(secreted frizzled related protein 1,SFRP1)和O6-甲基鸟嘌呤-DNA甲基转移酶(O6-methylguanine-DNA-methyltransferase,MGMT)基因启动子区DNA甲基化mRNA和蛋白在其中的表达和意义。方法CCK-8法检测不同浓度的5-Aza-CdR对人肺癌SPC-A1细胞增殖影响,划痕实验测定5-Aza-CdR对SPC-A1细胞迁移能力的影响,Hoechst 33258染色检测(0、3、10、30μmol·L^(-1))5-Aza-CdR处理肺癌SPC-A1细胞24h后细胞凋亡情况,RT-PCR、Western blot法检测SPC-A1细胞中SFRP1和MGMT的mRNA、蛋白表达。结果5-Aza-CdR可以浓度梯度的抑制肺癌SPC-A1细胞增殖,IC 50为21.2μmol·L^(-1);5-Aza-CdR(3、10、30μmol·L^(-1))作用48 h后,肺癌SPC-A1细胞划痕愈合分别为对照组的(92.4±2.6)%、(83.6±4.2)%、(76.7±4.5)%;5-Aza-CdR处理肺癌SPC-A1细胞后,出现典型的细胞凋亡形态学改变;不同浓度5-Aza-CdR(3、10、30μmol·L^(-1))处理SPC-A1细胞24 h后,SFRP1、MGMT mRNA和蛋白表达增加(P<0.05)。结论5-Aza-CdR可抑制肺癌SPC-A1细胞增殖,抑制肺癌细胞划痕修复和促进凋亡,可能与升高MGMT和SFRPl的甲基化表达有关。
Aim To observe the effect of methylation inhibitors of 5-aza-2′-deoxycytidine(5-Aza-CdR)on SPC-A1-lung cancer cell proliferation,cell scratches and apoptosis,to explore the influence of secretion curl associated protein 1(SFRP1)and O6-methylguanine-DNA-methyltransferase(MGMT)gene promoter region in which DNA methylation mRNA and protein expression and meaning.Methods The effects of 5-Aza-CdR at different concentrations on the proliferation of human lung cancer SPC-A 1 cells were determined by CCK-8 assay.The effect of 5-Aza-CdR on the migration ability of SPC-A 1 cells was determined by scratch assay.The apoptosis of lung cancer SPC-A 1 cells was detected by Hoechst 33258 staining after treatment with 5-Aza-CdR for 24 h.mRNA and protein expressions of SFRP1 and MGMT in SPC-A1 cells were detected by RT-PCR and Western blot.Results 5-Aza-CdR could reduce the proliferation of SPC-A 1 cells by concentration gradient,and IC 50 was 21.2μmol·L^(-1).After 5-Aza-CdR(3,10,30μmol·L^(-1))treated for 48 h,the lung cancer SPC-A 1 cell scratch healing rate was(92.4±2.6)%,(83.6±4.2)%,and(76.7±4.5)%,respectively.After 5-Aza-CdR treatment of lung cancer SPC-A 1 cells,there were typical morphological changes of apoptosis.After SPC-A 1 cells were treated with 5-Aza-CdR(3,10,30μmol·L^(-1))for 24 h,the expressions of mRNA and proteins of SFRP1 and MGMT increased(P<0.05).Conclusions 5-Aza-CdR can inhibit the proliferation of SPC-A 1 cells,inhibit cell scratch repair and promote the apoptosis of lung cancer cells,and to some extent promote the methylation of SFRP1and MGMT expressions.
作者
华丛书
叶静
陈海
葛腾飞
陈晓宇
HUA Cong-shu;YE Jing;CHEN Hai;GE Teng-fei;CHEN Xiao-yu(Dept of Thoracic Surgery,Chest Clinical Hospital of Anhui Medical University,Anhui Province Chest Hospital,Hefei 230022,China;Dept of Respiratory Medicine,the Second Hospital of Anhui Medical University,Hefei 230601,China;Experimental Center of Morphology,Anhui Medical University,Hefei 230032,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2021年第3期367-371,共5页
Chinese Pharmacological Bulletin
基金
国家自然科学资金面上项目(No 81373421)
安徽省胸科医院科研基金项目(No Tb15016)
安徽医科大学基础与临床合作提升计划项目(No 2019xkjT027)。
