ABSTRACT
Objective To explore the effect of invasion and migration of hepatocellular carcinoma (HCC) endothelial cells (TECs) affected by overexpression of microRNA-3178 (miR-3178) through the transfection of miR-3178 mimic.Methods Real-time polymerase chain reaction (Real-time PCR) was used to identify differential expression of miR-3178 in normal hepatic sinusoidal endothelial cells (HSECs) and HCC TECs.Furthermore,HCC TECs were divided into 3 groups:control (CON) group,miRNA-3178 upregulation (Mimics,up-regulation of miR-3178 expression was achieved using miR-3178 mimics transfected into HCC TECs) group and negative control (NC,negative control sequence was transfected into HCC TECs) group.RT-PCR was used to detect expression of miR-3178 in HCC TECs before and after transfection.Transfection efficiency was observed by using an inverted fluorescence microscope.HCC TECs invasionand migration were measured by matrigel invasion and transwell migration assay.EGR3 protein expression of HCC TECs were identified by Western blotting analysis.EGR3 mRNA expression of HCC TECs were identified by RT-PCR analysis.Results The results of RT-PCR showed that miR-3178 was significantly down-regulated in HCC TECs compared to HSECs (P ＜0.05),and expression of miR-3178 was significantly increased after the transcienttransfection (P ＜ 0.05).The transfection efficiency in HCC TECs was morethan 90％.Number of migrated and invaded cells and in miR-3178 group was significantly less than those in other groups.Target gene prediction software showed EGR3 was a possible candidate target.Transfection of miR-3178 mimic significantly decreased the mRNA and protein expression levels of EGR3.Conclusion MiR-3178 was downregulated in HCC TECs and overexpression of miR-3178 can specifically inhibit migration and invasion of HCC TECsin vitro through inhibiting EGR3 expression,thus,miR-3178 might be a critical targeted therapy strategv for HCC.
ABSTRACT
Objective To investigate the renal level of 1o-hydroxylase and the change of serum calcium in rats with severe acute pancreatitis,and their correlation.Methods Eighty male Wistar rats were randomly (random number) divided into two groups:sham operation group (SO group),severe acute pancreatitis group (SAP group),and each group was further randomly divided into 1 h,3 h,6 h,and 12 h subgroups (n =10).Severe acute pancreatitis model was made by retrograde infusion with 5％ sodium taurocholate solution into the biliopancreatic duct,rats were sacrificed at 1 h,3 h,6 h,and 12 h separately after modeling.The levels of serum amylase,serum calcium,serum urea nitrogen,serum creatinine,serum 1,25-dihydroxy vitamin D3 were measured,and the level of lα-hydroxylase protein in the kidney was determined with immunohistochemistry and Western blotting.The histopathologic changes of kidney tissue were observed under light microscope and the changes of the proximal tubular epithelial cell were observed under electron microscope.Results Compared with SO group,the levels of serum amylase,serum urea nitrogen,and serum creatinine were higher in SAP group,but the levels of serum calcium and 1,25-dihydroxy vitamin D3 decreased at 3,6,and 12 h,and the renal level of 1 α-hydroxylase also decreased at 3,6,and 12 h after modeling.In SAP group,the levels of serum calcium,serum 1,25-dihydroxy vitamin D3 and the renal level of 1 α-hydroxylase gradually decreased,and the renal level of 1 α-hydroxylase and the level of serum 1,25-dihydroxy vitamin D3 had positive correlation at 3 h,6 h,and 12 h (r =0.93,P ＜0.01; r=0.951,P ＜0.01; r =0.92,P ＜0.01; r =0.878,P ＜0.01),and the renal level of 1α-hydroxylase and the level of serum calcium had positive correlation at 3 h,6 h,and 12 h (r =0.975,P ＜0.01; r=0.946,P＜0.01; r=0.747,P＜0.01).Conclusions Intheearly course of SAP,the lowered activity of 1 α-hydroxylase may play an important role in the development of hypocalcemia.