ABSTRACT
Chronic granulomatous disease (CGD) is a rare type of primary phagocytic immunodeficiency disease.CGD is caused by a defective gene that encodes the components of reduced nicotinamide adenine dinucleotide phosphate (NADPH) in phagocytes.Due to genetic mutations causing phagocytic respiratory dysfunction,phagocytic cells are not effective in killing peroxidase-positive bacteria and fungi,and clinical manifestations are characterized by repeated severe bacterial,fungal infections and granuloma formation.The current clinical treatments include routine therapy and hematopoietic stem cell transplantation (HSCT).With the research development of CGD in genetics,molecular biology and vector science,clinical research on gene therapy of the disease has been carried out,in which CRISPR/Cas9 system has a good application prospect in gene therapy for genetic diseases.This article reviews the progress of gene therapy.
ABSTRACT
Objective:To investigate the anti-proliferation and anti-metastasis effects and study the molecular mechanism of sinomenine in cell line(HepG2).Methods: HepG2 cells were cultured together with different treatment concentrations of sinomen-ine.The effect of sinomenine on inhibition of growth of HepG2 cells were determined by methyl thiazolyl tetrazolium(MTT) assay.The effect of sinomenine on inhibiting metastasis of HepG2 cells were determined by Transwell assay.The inhibitory effect of sinomenine on reverse transcriptase(RT) was studied using inhibitory kinetic method,on the basis,the reactive oxygen species(ROS) of HepG2 cells was monitored by indirect fluorescent labeling.The protein expressions of CASP3,CASP9,CAV1 and SOX2 were analyzed by Western blot experiment.Results: Sinomenine inhibited the proliferation and metastasis of HepG2 cells significantly.Sinomenine had a good inhibitory effect on the growth of HepG2 cells,half inhibitory concentration(IC50) was (15.35±2.43)μmol/L.Sinomenine was RT inhibitor,IC50 was (21.32±2.43)μmol/L.The Western blot showed that CASP3,CASP9 and CAV1 were up-regulated and SOX2 was down-regulated by the sinomenine treatment in HepG2 cells.Conclusion: The potential molecular mechanism of sinomenine suppresses proliferation and metastasis of HepG2 cells by up-regulation of CASP3,CASP9,CAV1 and down-regulation of SOX2.