ABSTRACT
This article was aimed to investigate the cell proliferation , cell apoptosis and its related molecular mechanisms of the human gastric carcinoma cell line BGC-823 in v itro after treatment with cinnamaldehyde . The MTT Assay demonstrated the inhibitory effect of cinnamaldehyde . And the Flow Cytometry was used to determine its induction of cell apoptosis. The Hoechst 33342 was used to observe morphological changes during apoptosis . Moreover , quantitative real time PCR and western blot analysis were used to detect the effect of cinnamaldehyde on human gastric carcinoma cell line BGC-823 . The results showed that compared with the control group , cinnamaldehyde had inhibitory effect on human gastric carcinoma cell line BGC-823 ( P <0 . 01 ) . It showed that cinnamaldehyde induced apoptosis through the downregulation of Bcl-2 , Bcl-xL and Survivin expression , upregulation of Bax and Bak expression , downregulation of Bcl-2 and Procaspase-3 , and upregulation of BAX . It was concluded that cinnamaldehyde had inhibitory effect on the proliferation of human gastric carcinoma cell line BGC-823 and induced apoptosis . It may be related to the activation of the endogenous apoptosis pathway .
ABSTRACT
Objective:To construct specific small interfering RNA(siRNA) expressing vectors of intracellular Ca2+ transport protein(CaT1)gene and detect its silencing effects.Methods:The hairpin sequences of siRNAs targeting CaT1 gene were designed,synthesized and cloned into pSlincer 3.1-H1 plasmids after annealing.The vectors were then enriched in E.coli.The recombinant pSlincer 3.1-H1 plasmids were identified by restriction endonuclease cutting and DNA sequencing and then transfected into Human Gastric Carcinoma Cell Line BGC-823.The expression of CaT1 mRNA was examined by RT-PCR.Results:The siRNA oligonucleotides of CaT1 were correctly cloned into the pSlincer 3.1-H1 plasmids and confirmed by restriction endonuclease cutting and DNA sequencing.RT-PCR analysis revealed that the expression of CaT1 mRNA in BGC-823 cells transfected with the pSlincer 3.1-H1 constructs of siRNA was significantly decreased compared with that of the negative control and untransfected group.Conclusion:siRNA expression plasmids for silencing Ca2+ transport protein gene are successfully constructed,and they effectively inhibit the CaT1 gene expression.