摘要
目的探讨川陈皮素是否通过调控LINC00116影响结直肠癌细胞生物学功能。方法采用不同浓度川陈皮素处理结直肠癌细胞HCT116,采用MTT法和克隆实验检测细胞增殖与克隆形成,采用流式细胞仪分析细胞凋亡,采用Transwell小室法测定细胞的迁移和侵袭,采用Westernblotting法检测P21、含半胱氨酸的天冬氨酸蛋白水解酶3(Caspase-3)、E-钙黏蛋白(E-cadherin)、基质金属蛋白酶2(MMP-2)蛋白,采用实时荧光定量PCR检测LINC00116。在细胞HCT116中转染si-LINC00116,观察抑制LINC00116在细胞HCT116增殖、凋亡、迁移、侵袭中的作用。将pcDNA3.1-LINC00116转染入细胞HCT116,评估过表达LINC00116对川陈皮素作用的细胞HCT116增殖、凋亡、迁移、侵袭的影响。结果10、20、40μg/mL川陈皮素减少细胞HCT116的存活率、克隆形成数、迁移细胞数、侵袭细胞数、MMP-2蛋白表达、LINC00116表达,提高细胞凋亡率及P21、Caspase-3、E-cadherin蛋白表达,且作用效果随浓度增加而逐渐增强(P均<0.05)。抑制LINC00116降低细胞HCT116的细胞存活率、克隆形成数、迁移细胞数、侵袭细胞数、MMP-2蛋白表达,增加细胞凋亡率及P21、Caspase-3、E-cadherin蛋白表达(P均<0.05)。过表达LINC00116增加川陈皮素作用的细胞HCT116的细胞存活率、克隆形成数、MMP-2蛋白表达、迁移细胞数、侵袭细胞数,降低凋亡率及P21、Caspase-3、E-cadherin蛋白表达(P<0.05)。结论川陈皮素通过下调LINC00116的表达,抑制结直肠癌细胞的增殖、迁移、侵袭,并促进细胞凋亡。
Objective To study whether nobiletin affects the biological function of colorectal cancer cells by regulating long non-coding RNA(lncRNA)LINC00116.Methods Different concentrations of nobiletin were used to treat the colorectal cancer cells HCT116.The thiazole blue(MTT)and cloning were used to monitor cell proliferation and colony formation.Flow cytometry was used to analyze the apoptosis.Transwell chamber method was used to measure cell migration and cell invasion.Western blotting was used to detected the P21,Cysteinyl aspartate specific proteinase 3(Caspase-3),E-cadherin,and matrix metalloproteinase-2(MMP-2)proteins.Quantitative real-time PCR was used to detect the LINC00116.Si-LINC00116 transfected was into HCT116 cells,and we observed the roles of inhibiting LINC00116 on the proliferation,apoptosis,migration,and invasion of HCT116 cells.PcDNA3.1-LINC00116 was transfected into HCT116 cells,and we evaluated the effects of overexpression of LINC00116 on the cell proliferation,apoptosis,cell migration,and cell invasion of HCT116 cells treated with nobiletin.Results The 10,20 and 40μg/mL nobiletin reduced the survival rate,the number of colony formation,the number of migrating cells,the number of invading cells,the expression of MMP-2 protein,and the expression of LINC00116 in HCT116 cells,increased the apoptosis rate,P21,Caspase-3,and E-cadherin protein levels,and the effects gradually increased with the growing concentration(all P<0.05).Inhibition of LINC00116 reduced the cell survival rate,the number of colony formation,the number of migrating cells,the number of invading cells,and the expression of MMP-2 protein in HCT116 cells,and increased the apoptosis rate and the expression of P21,Caspase-3,and E-cadherin protein(all P<0.05).Overexpression of LINC00116 increased the cell survival rate,the number of clone formation,the expression of MMP-2 protein,the number of migrating cells,and the number of invading cells in HCT116 cells treated by nobiletin,which reduced the apoptosis rate,P21,Caspase-3,E-cadherin prot
作者
孙嘉伟
卢秀
王燕
SUNJiawei;LU Xiu;WANG Yan(The Fourth People's Hospital of Shandong Ji'nan,Ji'nan 250031,China;不详)
出处
《山东医药》
CAS
2021年第16期56-60,共5页
Shandong Medical Journal
