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Objective:To define the anti-gastric cancer activity in vitro of petroleum ether fraction of Boehmeria nivea root and reveal the material basis of its efficacy, so as to lay the foundation for the development and utilization of B. nivea root. Method:Methyl thiazolyl tetraolium(MTT) method was used to evaluate the inhibitory rate and time-dose relationship of petroleum ether fraction of B. nivea root with different doses and delivery times on human gastric cancer HGC-27 cells. Flow cytometry was used to detect the change of cell apoptosis and cell cycle after petroleum ether fraction of B. nivea root acted on human gastric cancer HGC-27 cells. GC-MS was used to detect the components of petroleum ether fraction of B. nivea root. Result:Experiment data showed significant cell proliferation inhibition in an obvious time-dose-effect manner, with statistically significant differences (PB. nivea root. The effect of petroleum ether fraction of B. nivea root on human gastric cancer HGC-27 cells could induce apoptosis,which affects the normal changes of cell cycle. The percentage of cells was decreased significantly in G0/G1 phase,and that in S phase was significantly increased. GC-MS was used to identify 26 chemical constituents in petroleum ether of B. nivea root,including sitosterol and stigmasterol. Conclusion:Petroleum ether fraction of B. nivea root is the active anti-gastric cancer part,and its main effective component is sterol compounds. This lays the foundation for the rational application of B. nivea root in clinic and the further research in tis anti-tumor effect.
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@# Objective: To observe the effect of ursolic acid (UA) on autophagy and apoptosis of gastric cancer cell line MGC-803, and to explore the mechanism of UA-induced autophagy of MGC-803 cells based on PI3K/AKT/mTOR signaling pathway. Methods: Human gastric cancer cell line MGC-803 was cultured in vitro and divided into blank control group, UAintervention group and UA+3-MA group. The cell apoptosis in each group was detected by flow cytometry. Cell autophagy was detected by double fluorescence mRFPeGFP-LC3 plasmid transfection method. The mRNA expression levels of LC3B, BAX and Bcl-2 were detected by qPCR. The protein expression levels of PI3K type I, p-AKT, p-mTOR, ULK1, LC3B, BAX and Bcl-2 were detected by WB. Results: Flow cytometry showed that the cell apoptotic rate of UA intervention group was significantly higher than that of blank control group (P<0.05). Compared with UAintervention group, the apoptotic rate in UA+ 3-MAgroup was significantly reduced (P<0.05). The double fluorescence mRFP-eGFP-LC3 plasmid transfection method showed that the green and red fluorescent bright spots in UA intervention group increased significantly compared with the blank control group (P<0.05), and the green and red fluorescent bright spots in UA+3-MA group were significantly reduced compared with UA intervention group (P<0.05). Real-time quantitative PCR and WB method showed that compared with the blank control group, the mRNAand protein expressions of BAX and LC3B, and ULK1 protein were significantly increased in UA intervention group, while the mRNA and protein expressions of Bcl-2, and the protein expressions of PI3K, p-AKT and p-mTOR were significantly decreased in UA intervention group (all P<0.05); Compared with UA intervention group, mRNA and protein expressions of BAX and LC3B were significantly down-regulated and the mRNAand protein expressions of Bcl-2 were significantly up-regulated in UA+3-MA group (all P<0.05), while protein levels of PI3K, p-AKT, p-mTOR and ULK1 were not significantly changed in UA+3-MA group (P>0.05). Conclusion: UA can promote apoptosis of MGC-803 cells via inducing autophagy, which may be related to UA's involvement in regulating the expressions of PI3K/AKT/mTOR signaling pathway-related proteins.
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ABSTRACT:Objective To investigate the effects of ω-3 polyunsaturated fatty acids on the proliferation of human gastric cancer cell line AGS and the possible mechanisms.Methods Human gastric cancer line AGS and human microvascular epithelial cell HMEC-1 were treated with different concentrations of docosdhexaenoic acid (DHA)and eicosapentaenoic acid (EPA).The inhibition of cell proliferation was evaluated by MTT assay and cell morphology.Flow cytometry was used to detect the cell cycle change.The expressions of mitochondrial respiratory membrane protein complex Ⅰ,Ⅱ and V were analyzed with Western blot.Results DHA and EPA could markedly inhibit the proliferation of AGS in significant time-dependent and concentration-dependent manners (P 0.05).Conclusion ω-3 PUFAs can selectively inhibit the growth and proliferation of human gastric cancer cell line AGS.These effects may be as-sociated with arresting cell cycle in G0/G1 phase and inhibiting the energy metabolism of AGS cells.
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OBJECTIVE: To investigate the effect of galangin on the anti-proliferation and apoptosis of gastric cancer SGC-7901 cells, and possible mechanisms. METHODS: Proliferative activity of SGC-7901 cells was measured by MTT assay while that cell cycle progression and apoptosis of galangin-treated SGC-7901 cells were analyzed by flow cytometry, morphologic observation and mitochondrial membrane potential (MMP) analysis. RESULTSS The evaluated results showed that when galangin was added into cell culture in 40-200 μmol · L-1, proliferative inhibition occurred and was observed as a dose- and time-dependent manner. The calculated IC50 values for galangin were 160, 100 and 70 μmol · L-1 when the treatment time was 24, 48 and 72 h, respectively. Flow cytometry analysis revealed that the proportion of the cells in the G2/M phase was significantly enhanced from 4.40(control group) to 18.31 (treatment group) by a treatment time of 24 h. CONCLUSION: The cells in treatment group showed typical apoptotic morphology and a decrease in MMP. At the same time, the percentage of the apoptotic cells significantly increased from 2.6% or 4.3% (control group) to 27.4% or 65.6% (treatment group), when 160 μmol · L-1 galangin treated SGC-7901 cells for 24 or 48 h. These mentioned results indicate that galangin can inhibit the proliferation and induce apoptosis of the SGC-7901 cells by perturbation in cell cycle progression and mitochondrial dysfunction.
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This study was aimed to investigate effect of aconite alkaloids on proliferation and apoptosis of hu-man gastric cancer cell line SGC-7901 . Effects of different concentrations of aconite alkaloids on proliferation of human gastric cancer cell line SGC-7901 were investigated with MTT assay; induced apoptosis and cell cy-cle blocking were detected with flow cytometer ( FCM ) . The results showed that the IC50 of aconite alkaloids on human gastric cancer cell line SGC-7901 was 0 . 2318 ± 0 . 0022 , 0 . 1601 ± 0 . 0227 , 0 . 1031 ± 0 . 0231 mg/ml at 24 h , 48 h and 72 h , respectively , compared to the control group with significant difference ( P <0.01). When the aconite alkaloids concentration was 0.8 mg/ml, it appeared with obvious apoptosis. The apop-tosis rate was ( 59 . 38 ± 5 . 05 )%. The FCM detection showed that compared with control group , the percentage of S-phase cells increased in the treatment group . And a typical sub-diploid peak appeared before G0 / G1 phase . It was concluded that aconite alkaloids can inhibit the proliferation of human gastric cancer cell line SGC-7901 in vitro, induce the apoptosis of cells and make SGC-7901 cells remain in S phase.
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We used retrovial vector LXSN to construct recombinant retroviral vector with mutant membrane bound TNF - ? gene. The vectors were introduced into packaging cell line, CRIP cells. The G418-resistant colonies were selected and the supernatants of the colonies were used to determine the virus titers. The titer of virus was 1? 104CFU/ml and the retroviral vectors were used to tranduced the human gastric cancer cell line, MGC-803 cells. The results of southern blot assay showed thai the targel gene had integrated into the genomic DNA of MGC-803T. MGC - 803Tcells were ablc to kill L929 cell line, but the parent cell line showed no cytotoxicily to the cells at all. There was no any variance in the morphological appearance and growlh rate in vilro of MGC-803N and MGC-803T cells. The results of inoculation in nude mice with these cells indicated that MGC-803T cells showed a considerable decrease in size of tumor. These results suggested that the retroviral vectors expressing mulant TNF -? gene were successfully construed. MGC - 803T cells showed cytotoxicily slrongly lo L929 cell line in vilro and lumorigenicity .