榄香烯通过下调NCOA2表达抑制胃癌细胞增殖、侵袭、迁移的实验研究

Experimental Study on Elemene Inhibiting the Proliferation,Invasion and Migration of Gastric Cancer Cells by Down-regulating the Expression of NCOA2

目的:探究榄香烯对核受体辅激活因子2(NCOA2)表达及胃癌细胞增殖、侵袭、迁移的影响。方法:用不同浓度的榄香烯处理人胃腺癌BGC-823细胞,实时荧光定量PCR(qRT-PCR)和蛋白印迹法(Western blot)检测NCOA2 mRNA和蛋白的相对表达量。再次培养BGC-823细胞,将细胞分为对照组(不做任何处理)、榄香烯组(0.08 mg·ml^(-1)榄香烯),随后在榄香烯组的基础上再分为阴性对照组(转染pcDNA空载质粒)和榄香烯+NCOA2组(转染pcDNA-NCOA2过表达质粒)。平板克隆实验及MTT法分别检测各组细胞的集落形成率及增殖活力;划痕实验及Transwell实验分别检测各组细胞的划痕愈合率及侵袭细胞数量;Western blot检测各组细胞中NCOA2及增殖、侵袭、迁移相关蛋白水平。结果:与榄香烯0 mg·ml^(-1)相比,不同浓度的榄香烯可明显下调NCOA2的mRNA和蛋白表达水平,且0.02~0.08 mg·ml^(-1)范围内,NCOA2表达随着榄香烯浓度的增高而降低(P<0.05)。与对照组相比,榄香烯组BGC-823细胞的集落形成率、增殖活力、划痕愈合率、侵袭细胞数量以及NCOA2、细胞周期蛋白CyclinD1、增殖细胞核抗原(PCNA)、N-钙黏蛋白(N-Cadherin)及波形蛋白(Vimentin)蛋白水平显著降低,E-Cadherin蛋白水平显著增高(P<0.05);与阴性对照组相比,榄香烯+NCOA2组细胞集落形成率、增殖活力、划痕愈合率、侵袭细胞数量以及NCOA2、CyclinD1、PCNA、N-Cadherin及Vimentin蛋白水平明显升高,E-Cadherin蛋白水平明显降低(P<0.05);榄香烯组与阴性对照组相比,各项指标则无显著差异(P>0.05)。结论:榄香烯可通过下调NCOA2表达进而抑制胃癌细胞增殖、侵袭及迁移。

Objective:To explore the effects of elemene on the expression of nuclear receptor coactivator 2(NCOA2) and the proliferation,invasion and migration of gastric cancer cells.Methods:Human gastric adenocarcinoma BGC-823 cells were treated with different concentrations of elemene,and the relative expression of NCOA2 mRNA and protein was detected by real-time fluorescent quantitative PCR(qRT-PCR) and Western blot.The BGC-823 cells were cultured again and divided into control group(without any treatment) and elemene group( treated with 0.08 mg·ml^(-1) elemene),and then the elemene group was divided into negative control group(transfection of pcDNA empty plasmid) and elemene+NCOA2 group(transfection of pcDNA-NCOA2 overexpression plasmid).Plate cloning experiment and MTT method were used to detect the colony formation rate and proliferation activity of each group of cells,respectively;scratch test and Transwell test were used to detect the scratch healing rate and the number of invaded cells of each group of cells,respectively;Western blot was used to detect the levels of NCOA2 and the proliferation,invasion and migration related proteins in each group of cells.Results:Compared with elemene 0 mg·ml^(-1),different concentrations of elemene could significantly down-regulate the expression levels of NCOA2 mRNA and protein,and within the range of 0.02-0.08 mg·ml^(-1),the expression of NCOA2 increased with the concentration decrease of elemene(P<0.05).Compared with those in the control group,the colony formation rate,proliferation activity,scratch healing rate,number of invaded cells,NCOA2,CyclinD1,proliferating cell nuclear antigen(PCNA),N-cadherin and Vimentin protein levels of BGC-823 cells in elemene group were significantly reduced,while E-Cadherin protein level was significantly increased(P<0.05);compared with those in the negative control group,the colony formation rate,proliferation activity,scratch healing rate,number of invaded cells,NCOA2,CyclinD1,PCNA,N-cadherin and Vimentin protein levels in elemene+NCOA2 group were significantly increased,while E-Cadherin protein level was significantly reduced(P<0.05);however,there were no significant differences in the cell indicators between elemene group and the negative control group(P>0.05).Conclusion:Elemene can inhibit the proliferation,invasion and migration of gastric cancer cells by down-regulating the expression of NCOA2.