Objective To investigate the effects of microRNA-18a(miR-18a) on migration and invasion of hepatocellular carcinoma(HCC) cells,and its possible mechanism associated with Dicer l. Methods HepG 2 and HepG 2.2.15 cells w...Objective To investigate the effects of microRNA-18a(miR-18a) on migration and invasion of hepatocellular carcinoma(HCC) cells,and its possible mechanism associated with Dicer l. Methods HepG 2 and HepG 2.2.15 cells were transfected with miR-18 a inhibitor using Lipofectamine. Cell invasion was evaluated by transwell invasion assay,and cell migration was detected by transwell migration and wound-healing assays. Moreover,luciferase reporter assay was used to identify whether Dicer expression was regulated by miR-18 a. Real-time RT-PCR and western blot were performed to analyze Dicer 1 expression. In addition,a functional restoration assay was performed to investigate whether miR-18 a promotes HCC cell migration and invasion by directly targeting Dicer 1. Results miR-18 a inhibitor can suppress the migration and invasion of HCC cells. Furthermore,suppression of Dicer l expression by small interfering RNA essentially abolished the inhibition of cell migration and invasion induced by miR-18 a inhibitor,restorating these activities to levels similar to the parental HCC cells. Interestingly,suppression of miR-18 a in HCC cells resulted in enhanced expression of Dicer l. In addition,the results of a luciferase assay demonstrated targeted regulation of Dicer l by miR-18 a. Conclusion Our findings suggest that miR-18 a promotes migration and invasion of HCC cells by inhibiting Dicer l expression.展开更多
磷酸酶互作蛋白2(Phosphatase-interacting protein 2,PSTPIP2)和溶质载体家族6成员18(Solute carrier family 6 member 18,SLC6A18)基因在动物先天性自身免疫中发挥着重要的作用。为探讨PSTPIP2基因g.43165308C>T位点和SLC6A18基因g...磷酸酶互作蛋白2(Phosphatase-interacting protein 2,PSTPIP2)和溶质载体家族6成员18(Solute carrier family 6 member 18,SLC6A18)基因在动物先天性自身免疫中发挥着重要的作用。为探讨PSTPIP2基因g.43165308C>T位点和SLC6A18基因g.8294767T>C位点多态性与牦牛免疫性状的关联性,本研究对189头娘亚牦牛的PSTPIP2基因g.43165308C>T位点和SLC6A18基因g.8294767T>C位点进行了PCR扩增,并利用PARMS SNP分型,采用IBM SPSS Statistics 26.0软件分析了不同基因位点与牦牛免疫球蛋白、炎症标志物及细胞因子等免疫性状的关联性。结果表明,PSTPIP2基因g.43165308C>T位点存在CC、CT和TT三种基因型,优势等位基因和基因型分别是C和CC,其频率分别为0.720和0.534。PSTPIP2基因g.43165308C>T位点属于中度多态位点,且处于Hardy-Weinberg平衡状态(P>0.05)。SLC6A18基因g.8294767T>C位点存在CC、TC和TT三种基因型,优势等位基因和基因型分别是T和TT,其频率分别为0.796和0.660。SLC6A18基因g.8294767T>C位点属于中度多态位点,且处于Hardy-Weinberg不平衡状态(P<0.05)。关联分析结果表明,PSTPIP2基因g.43165308C>T位点与牦牛免疫球蛋白A(IgA)、免疫球蛋白M(IgM)、C反应蛋白(CRP)、白介素-2(IL-2)、白介素-4(IL-4)、白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)均呈显著相关(P<0.05);SLC6A18基因g.8294767T>C位点与C反应蛋白(CRP)、白介素-4(IL-4)、干扰素-γ(IFN-γ)均呈显著相关(P<0.05)。综上所述,PSTPIP2和SLC6A18基因的多态性与牦牛免疫性状显著相关,这两个基因可能是影响牦牛免疫性状的潜在候选基因。展开更多
基金Supported by the National Natural Science Foundation of China(81472485)the Project of Six Talent Peaks of Jiangsu Province(WSW-014)Project of Hospital Management Center of Wuxi(YGZXQ1311,YGZ1106,and YGZ1007)
文摘Objective To investigate the effects of microRNA-18a(miR-18a) on migration and invasion of hepatocellular carcinoma(HCC) cells,and its possible mechanism associated with Dicer l. Methods HepG 2 and HepG 2.2.15 cells were transfected with miR-18 a inhibitor using Lipofectamine. Cell invasion was evaluated by transwell invasion assay,and cell migration was detected by transwell migration and wound-healing assays. Moreover,luciferase reporter assay was used to identify whether Dicer expression was regulated by miR-18 a. Real-time RT-PCR and western blot were performed to analyze Dicer 1 expression. In addition,a functional restoration assay was performed to investigate whether miR-18 a promotes HCC cell migration and invasion by directly targeting Dicer 1. Results miR-18 a inhibitor can suppress the migration and invasion of HCC cells. Furthermore,suppression of Dicer l expression by small interfering RNA essentially abolished the inhibition of cell migration and invasion induced by miR-18 a inhibitor,restorating these activities to levels similar to the parental HCC cells. Interestingly,suppression of miR-18 a in HCC cells resulted in enhanced expression of Dicer l. In addition,the results of a luciferase assay demonstrated targeted regulation of Dicer l by miR-18 a. Conclusion Our findings suggest that miR-18 a promotes migration and invasion of HCC cells by inhibiting Dicer l expression.
文摘磷酸酶互作蛋白2(Phosphatase-interacting protein 2,PSTPIP2)和溶质载体家族6成员18(Solute carrier family 6 member 18,SLC6A18)基因在动物先天性自身免疫中发挥着重要的作用。为探讨PSTPIP2基因g.43165308C>T位点和SLC6A18基因g.8294767T>C位点多态性与牦牛免疫性状的关联性,本研究对189头娘亚牦牛的PSTPIP2基因g.43165308C>T位点和SLC6A18基因g.8294767T>C位点进行了PCR扩增,并利用PARMS SNP分型,采用IBM SPSS Statistics 26.0软件分析了不同基因位点与牦牛免疫球蛋白、炎症标志物及细胞因子等免疫性状的关联性。结果表明,PSTPIP2基因g.43165308C>T位点存在CC、CT和TT三种基因型,优势等位基因和基因型分别是C和CC,其频率分别为0.720和0.534。PSTPIP2基因g.43165308C>T位点属于中度多态位点,且处于Hardy-Weinberg平衡状态(P>0.05)。SLC6A18基因g.8294767T>C位点存在CC、TC和TT三种基因型,优势等位基因和基因型分别是T和TT,其频率分别为0.796和0.660。SLC6A18基因g.8294767T>C位点属于中度多态位点,且处于Hardy-Weinberg不平衡状态(P<0.05)。关联分析结果表明,PSTPIP2基因g.43165308C>T位点与牦牛免疫球蛋白A(IgA)、免疫球蛋白M(IgM)、C反应蛋白(CRP)、白介素-2(IL-2)、白介素-4(IL-4)、白介素-6(IL-6)、肿瘤坏死因子-α(TNF-α)均呈显著相关(P<0.05);SLC6A18基因g.8294767T>C位点与C反应蛋白(CRP)、白介素-4(IL-4)、干扰素-γ(IFN-γ)均呈显著相关(P<0.05)。综上所述,PSTPIP2和SLC6A18基因的多态性与牦牛免疫性状显著相关,这两个基因可能是影响牦牛免疫性状的潜在候选基因。
