以地被菊(Chrysanthemum morifolium Ramat.)'七月桃花'为材料,利用同源基因克隆法结合RACE技术克隆了光周期调控开花重要基因FLOWERING LOCUS T(FT)的类似基因(基因登陆号GQ925916,命名为CmFT)。该基因开放阅读框有525bp,可编...以地被菊(Chrysanthemum morifolium Ramat.)'七月桃花'为材料,利用同源基因克隆法结合RACE技术克隆了光周期调控开花重要基因FLOWERING LOCUS T(FT)的类似基因(基因登陆号GQ925916,命名为CmFT)。该基因开放阅读框有525bp,可编码174个氨基酸。蛋白比对发现,CmFT所推测的氨基酸序列包含FT类蛋白保守基序和两个关键性氨基酸残基。通过系统遗传进化树分析进一步表明CmFT属于FT亚家族成员。SYBR GREEN法实时荧光定量RT-PCR表达分析结果表明CmFT在花芽的表达量最高,茎的表达量次之,叶片表达量最低。在短日条件下,该基因的表达量呈昼夜节律表达型,在光照期间呈下降趋势,在夜间逐渐上升,光照前4h时达到最高水平。在长日条件下,其相对表达量显著低于短日条件下且趋于零。由此推测,该基因主要与菊花光周期敏感性密切相关,可能在光周期促进开花中发挥一定的作用。展开更多
Objective To explore the role and regulation of guanine nucleotide-binding protein G（i）, a-1 subunit （GNAI1） in hepatocellular carcinoma （HCC）. Methods Expression of GNAI1 in HCC samples was determined by qRT-PC...Objective To explore the role and regulation of guanine nucleotide-binding protein G（i）, a-1 subunit （GNAI1） in hepatocellular carcinoma （HCC）. Methods Expression of GNAI1 in HCC samples was determined by qRT-PCR and immunohistochemical （IHC） staining. Huh-7 and SNU-387 cells stably expressing GNAI1 were established by the infection of lentivirus transducing unit containing GNAI1. siRNA against GNAI1 was transfected into SMMC-7721 cells to knock down the GNAI1 expression in HCC cells. Mir-320a/c/d mimics were transfected into SMMC-7721 and SK-Hep-1 cells and the expression of GNAll was determined by Western blot. The migration and invasion of Huh-7, SNU-387, SK-Hep-1 and SMMC-7721 cells were investigated by Transwell assays. Results The GNAI1 protein was significantly downregulated in HCC samples without changes in its mRNA levels. GNAI1 could inhibit the migration and invasion of HCC cells in vitro. Further investigations indicated that GNAI1 was a target of miR-320a/c/d in HCC cells. Transwell assays demonstrated that these microRNAs could promote the migratory ability and invasivesess of HCC cells in vitro. Conclusions GNAII is downregulated in HCC and inhibits the migration and invasion of HCC cells. This study is the first to investigate the role of GNAI1 in cancer. Regulation of GNAI1 by miR-320a/c/d indicates new therapeutic avenues for targeting HCC metastasis.展开更多
基金supported by grants from the National 973 Key Basic Research Program(No.2011CB933100)National Natural Science Foundation of China(No.81125016 and 81101481)+1 种基金Science and Technology Commission of Shanghai Municipality(No.11XD1404500 and 10JC1414200)Shanghai Health Bureau(No.XYQ2011047 and XBR2011039)
文摘Objective To explore the role and regulation of guanine nucleotide-binding protein G（i）, a-1 subunit （GNAI1） in hepatocellular carcinoma （HCC）. Methods Expression of GNAI1 in HCC samples was determined by qRT-PCR and immunohistochemical （IHC） staining. Huh-7 and SNU-387 cells stably expressing GNAI1 were established by the infection of lentivirus transducing unit containing GNAI1. siRNA against GNAI1 was transfected into SMMC-7721 cells to knock down the GNAI1 expression in HCC cells. Mir-320a/c/d mimics were transfected into SMMC-7721 and SK-Hep-1 cells and the expression of GNAll was determined by Western blot. The migration and invasion of Huh-7, SNU-387, SK-Hep-1 and SMMC-7721 cells were investigated by Transwell assays. Results The GNAI1 protein was significantly downregulated in HCC samples without changes in its mRNA levels. GNAI1 could inhibit the migration and invasion of HCC cells in vitro. Further investigations indicated that GNAI1 was a target of miR-320a/c/d in HCC cells. Transwell assays demonstrated that these microRNAs could promote the migratory ability and invasivesess of HCC cells in vitro. Conclusions GNAII is downregulated in HCC and inhibits the migration and invasion of HCC cells. This study is the first to investigate the role of GNAI1 in cancer. Regulation of GNAI1 by miR-320a/c/d indicates new therapeutic avenues for targeting HCC metastasis.
