姜黄素对肺癌细胞TFPI-2基因去甲基化作用研究  

Study on the Effect of Curcumin on the Demethylation of TFPI-2 Gene in Lung Cancer Cells

作　　者：梁莹[1] 吴西雅[1] 肖祖克[1] LIANG Ying;WU Xi-ya;XIAO Zu-ke(Jiangxi Provincial People's Hospital,Nanchang,Jiangxi,China,330000)

机构地区：[1]江西省人民医院,江西南昌330000

出　　处：《河南中医》2022年第12期1852-1856,共5页Henan Traditional Chinese Medicine

基　　金：江西省中医药管理局科技计划项目(2020B0337)。

摘　　要：目的:探讨姜黄素对肺癌细胞组织因子途径抑制物-2(tissue factor pathway inhibitor-2,TFPI-2)基因去甲基化的作用。方法:将人A549细胞分为姜黄素低剂量组(20μmol·L^(-1))、姜黄素中剂量组(40μmol·L^(-1))、姜黄素高剂量组(80μmol·L^(-1))及空白对照组,分别采用完全培养液及含相应浓度姜黄素的培养液培养后,MTT法检测A549细胞的增殖情况;RT-PCR检测TFPI-2 mRNA及DNA甲基转移酶1(DNA methyltransferase 1,DNMT1)mRNA、DNMT3a mRNA、DNMT3b mRNA的表达情况;甲基化特异性PCR(methylation-specific PCR,MSP)法检测TFPI-2基因甲基化状态。结果:与空白对照组比较,处理细胞12 h、24 h及48 h后,各剂量姜黄素组细胞增殖降低,且40μmol·L^(-1)姜黄素处理细胞48 h时OD值最低,差异有统计学意义(P<0.05)。细胞抑制率结果显示,各剂量姜黄素处理细胞12 h、24 h及48 h后均可抑制细胞增殖。与空白对照组比较,姜黄素各剂量组TFPI-2 mRNA表达水平升高,DNMT1 mRNA、DNMT3a mRNA、DNMT3b mRNA表达水平降低,差异均具有统计学意义(P<0.05)。TFPI-2甲基化检测显示:空白对照组未显示甲基化及去甲基化产物,各剂量姜黄素组同时显示甲基化产物和去甲基化产物,尤以姜黄素中剂量组去甲基化产物条带最为明显。结论:姜黄素可通过降低DNMT1 mRNA、DNMT3a mRNA、DNMT3b mRNA表达水平抑制TFPI-2甲基化,进而抑制A549细胞增殖,且以浓度40μmol·L^(-1)抑制效果最佳。Objective:To study the effect of curcumin on the demethylation of tissue factor pathway inhibitor-2(TFPI-2)gene in lung cancer cells.Methods:A549 cells were divided into the low-dose curcumin group(20μmol·L^(-1)),the medium-dose curcumin group(40μmol·L^(-1)),the high-dose curcumin group(80μmol·L^(-1))and the blank control group.Then they were cultured with ordinary culture medium and culture medium containing curcumin of corresponding concentration respectively,and the proliferation of A549 cells was detected by MTT method.The expressions of TFPI-2 mRNA,DNA methyltransferase 1(DNMT1)mRNA,DNMT3 a mRNA and DNMT3 b mRNA were detected by RT-PCR.Methylation specific PCR(MSP)was used to detect the methylation status of TFPI-2 gene.Results:Compared with the blank control group,the proliferation of cells in curcumin groups with different doses decreased after treatment of 12 hours,24 hours and 48 hours respectively,and the OD value of cells treated with 40μmol·L^(-1) of curcumin for 48 hours was the lowest,and the difference was statistically significant(P<0.05).The results of cell inhibition rate showed that curcumin could inhibit cell proliferation after 12 hours,24 hours and 48 hours respectively.Compared with the blank control group,the expression level of TFPI-2 mRNA in each dose group of curcumin increased,and the expression levels of DNMT1 mRNA,DNMT3 a mRNA,and DNMT3 b mRNA decreased,and all the differences were statistically significant(P<0.05).TFPI-2 methylation detection showed that the blank control group did not show methylation and demethylation products,and all dose curcumin groups showed both methylation and demethylation products,especially in the medium-dose curcumin group.Conclusion:Curcumin can inhibit TFPI-2 methylation by reducing the expression levels of DNMT1 mRNA,DNMT3 a mRNA,and DNMT3 b mRNA,thereby inhibiting the proliferation of A549 cells.And at the concentration of 40μmol·L^(-1),the inhibition effect was the best.

关 键 词：肺癌 姜黄素 组织因子途径抑制物-2 去甲基化 

分 类 号：R285.5[医药卫生—中药学]