ABSTRACT
【Objective】 To investigate the effects of RASSF6 gene on gastric cancer cells’ proliferation， autophagy， apoptosis， and sensitivity to oxaliplatin chemotherapy. 【Methods】 Gastric cancer BGC823 cells were cultured in vitro and divided into experimental control group （control group）， RASSF6 overexpression group （Oe group）， RASSF6 interference group， and lentivirus control group according to the expression effect of lentivirus gene. The changes in cell proliferation， cell cycle distribution， cell migration， autophagy， apoptosis and sensitivity to oxaliplatin in each group were detected， and the number of autophagy bodies in each group was detected by electron microscopy. Real-time PCR （qRT-PCR） and Western blotting were used to detect the expression levels of apoptosis- and autophagy-related genes in each group. 【Results】 Studies on the biological behavior of gastric cancer BGC823 cells induced by RASSF6 gene expression showed that compared with the control group， the percentage of G0/G1 phase cells in the Oe group increased， while the percentage of G2 and S phase cells decreased， with statistical significance （P<0.05）. The apoptosis rate was significantly increased （P<0.05）. The cell scratch assay showed that the scratch healing rate was significantly decreased （P<0.05）. Studies on the sensitivity of RASSF6 gene expression to oxaliplatin showed that compared with the drug group （L-OHP group）， the apoptosis rate of Oe+L-OHP group was increased significantly （P<0.05）. In the Oe+L-OHP group， the expression of anti-apoptotic protein Bcl-2 decreased， the expressions of Bax and Caspase-3 were increased； the expression of autophagosomes was increased； the expressions of Beclin-1 and P62 and the ratio of LC3-Ⅱ/LC3 were all increased （P<0.05）. 【Conclusion】 The RASSF6 gene plays a role in suppressing gastric cancer cell BGC823， which can increase the sensitivity to oxaliplatin chemotherapy by promoting apoptosis and autophagy.
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.
ABSTRACT
Ten cell strains were obtained from the human gastric cancer cell line BGC-823 by using the method of single cell cloning. Through inoculation of these cell strains in nude mice and subsequent observation of their metastatic capability, four cell lines, C_1, C_6 with high, and C_5, C_8 with low metastatic capability were selected out; and their biological and morphological properties were studied. It was demonstrated that all of them did not show apparent difference in growth ability, but morphologically, the mictovilli of C_5 and C_8 cells were scarce and short; and their microfilaments were thick, straight and well organized. Whereas the microvilli of C_1 and C_6 strains were long and abundant comparatively, but their microfilaments were poorly organized. On the basis of present observation,we suggested that the organization of microfilaments in cancer cells appeared to bear reversed relationship with their metastatic capability.