E3 Ubiquitin Ligase HRD1 On The Uptake Rate Of ¹⁸F-FDG In Hepatoma Cells

Background ¹⁸F-FDG(¹⁸F-fluoro-2-deoxy-glucose),an analogue of glucose,can be used as substrates for energy metabolism and tumor PET-CT imaging tracer.Hepatocellular carcinoma on¹⁸F-FDG uptake in the degree of differentiation and differentiation of the same cells showed great difference.Density distribution which depends mainly on the cell membrane of GLUT1.GLUT1 is the main effect of glucose uptake and transport of tumor cells ¹⁸F-FDG protein.This study,according to the post-translational modification of protein synthesis after endoplasmic reticulum stress related protein degradation in molecular mechanism,focuses on the research of cytokines in liver cells with IL-6 mediated by ubiquitin E3 ligase HRD1 mediated endoplasmic reticulum degradation of GLUT1 protein,the cell membrane of GLUT1 distribution to reduce the use of sugar,weaken the function of the effect of ¹⁸F-FDG uptake.Culture experiments and animal model experiment method in class group by Huh7 cells in vitro,clinical cases of hepatocellular carcinoma cell line,the expression of GLUT1 and the regulation effect of HRD1 modified IL-6 molecular level.Based on preclinical work,provide the basis for unraveling the molecular mechanisms underlying HCC ¹⁸F-FDG PET-CT uptake imaging differences.Object To investigate the molecular mechanism of PET-CT imaging difference of ¹⁸F-FDG in HCC.We hypothesized that HRD1,a E3 ubiquitin ligase,mediate degradation of GLUT1 protein,result in PET-CT imaging difference of ¹⁸F-FDG in HCC.Methods 1.Analysis the ¹⁸F-FDG uptake,the expression of GLUT1,HRD1 and Derlin1 in 9 HCC patients by PET-CT imaging detected by immunofluorescence assay and immunohistochemistry assay.Detect GLUT1 protein and m RNA expression of in HCC carcinoma,paracancerous and normal tissues by Western blot and RT-PCR.2.Examine the effect of IL-6 on the protein expression of HRD1 in Huh7 cells in vitro.3.Construction of HRD1 overexpression and HRD1-Sh RNA Huh7 cell line in vitro.4.Detection of¹⁸F-FDG uptake rate in HRD1 Sh RNA Huh7 and HRD1 overexpression cell lines.5.Construction of HRD1 Sh RNA Huh7 and HRD1 overexpression Balb-c in nude mice model.Detection of¹⁸F-FDG uptake rate of different groups by Micro PET in vivo 6.Detect of the protein expression of GLUT1,HRD1 and Derlin1 in different groups by western blot.7.Detect the interaction between HRD1 and GLUT1 by CO-IP.Results 1.We collect 9 HCC patients with primary lesions in the left liver lobe or right lobe,solitary,irregular shape and unclear boundary.PET-CT results indicate that length of the lesion size is about 2.0-13 cm,the average value of CT 35-55 HU,SUVmax 2.5-10.2.These patients can be divided into three groups by SUVmax,including low uptake group?3 cases,SUVmax <3?,moderate uptake group(3 cases,3<SUVmax...