Objective:To explore the influence of silencing Bcl-2 expression by small interfering RNA（siRNA） on Bcl-2 protein expression,cell apoptosis rale and radiosensilivity of gastric cancer BCC823 cells.Methods:siRNA segm...Objective:To explore the influence of silencing Bcl-2 expression by small interfering RNA（siRNA） on Bcl-2 protein expression,cell apoptosis rale and radiosensilivity of gastric cancer BCC823 cells.Methods:siRNA segment for Bcl-2 gene was designed and synthesized,then was induced into gastric cancer BGC 823 cells by liposome transfection.Bcl-2 protein expression was detected by Western Blotting.After X radiation,flow cytometry and clone forming assay were used to determine the effects of RNA interference on BGC823 cell apoptosis rate and radiosensitivity. Result:After the transfection of Bcl-2 siRNA,the positive expression rate of Bcl-2 protein in BGC823 cells was（35.45±2.35）%.Compared with the control group and negative siRNA transfection group,the rate was significantly decreased（P【0.01）.The apoptosis rate of BGC823-RNAi cell was（10.81±0.91）%,which was significantly higher than the control group and negative siRNA transfection group（P【0.01）.After 48h X radiation,the apoptosis rate of BGC823-RNAi was（28.91±1.40）%,which was significantly higher than the control group and the group without radiation （P【0.01）.During clone forming assay D<sub>0</sub>,D<sub>4</sub> and SF<sub>2</sub> values in Bcl-2 siRNA1 transfection group were all lower than those in the control group.The radiosensitivity ratio was 1.28（the ratio of D<sub>0</sub>） and 1.60（the ratio of D<sub>4</sub>）.Conclusions:Specific siRNA of Bcl-2 gene can effectively inhibit the expression of Bcl-2 gene,enhance the radiosensitivity and apoptosis of gastric cancer BGC823 cells,having good clinical application perspective.展开更多
[Objectives]This study aimed to investigate the inhibiting effect of Obazema on proliferation of gastric cancer BGC-823 cells and its mechanism.[Methods]BGC-823 cells were treated with high,medium and low concentratio...[Objectives]This study aimed to investigate the inhibiting effect of Obazema on proliferation of gastric cancer BGC-823 cells and its mechanism.[Methods]BGC-823 cells were treated with high,medium and low concentrations of drug-containing serum(0.316%,0.158%and 0.079%)for 0,48,72 and 96 h,respectively.Then,the proliferation of the cells was detected with CCK-8 method,and the expression of related proteins,B lymphocytoma-2(Bcl-2),phosphorylated protein kinase B(p-Akt),protein kinase B(Akt)and glyceraldehyde-3-phosphate dehydrogenase(GAPDH),was detected using Western blotting.[Results]The proliferation of the BGC-823 cells was significantly inhibited with different doses of Boenninghausenia albiflora(Hook.)Reichb.ex Meisn.var.albiflora(CH)and B.sessilicarpa Lévl.(S)(P<0.01),in dose-dependent and time-dependent manners.The inhibition of high-dose S on cell proliferation was similar to that of CTX 48 h after administration;the inhibition of high-dose CH on cell proliferation was significantly stronger than that of CTX(P<0.01);different doses of drug administration groups significantly inhibited the expression of p-Akt and Bcl-2 in the BGC-823 cells;the inhibition of high-dose CH on the expression of P-Akt and Bcl-2 and the inhibition of medium-dose CH on the expression of Bcl-2 were significantly stronger than that of CTX(P<0.05,P<0.01),in a certain dose-dependent manner;at the same dose,the inhibition of CH on the expression of the proteins was stronger than that of S(P<0.05,P<0.01);administration of S and CH significantly inhibited the expression of GAPDH compared with CTX(P<0.05,P<0.01).[Conclusions]Obazema has the capacity to inhibit the proliferation of BGC-823 cells.The mechanism may be achieved by inhibiting the expression of p-Akt and Bcl-2,and GAPDH may be the target gene of its anti-tumor mechanism.The inhibiting effect of CH on BGC-823 cells was more significantly than that of S.展开更多
The effect of LaCit on cell proliferation, DNA synthesize, cell cycle, cell membrane fluidity, growth in soft agar in human lung cancer cells PG and human gastric carcinoma cells BGC 823 was studied by MTT(3 (4,5 d...The effect of LaCit on cell proliferation, DNA synthesize, cell cycle, cell membrane fluidity, growth in soft agar in human lung cancer cells PG and human gastric carcinoma cells BGC 823 was studied by MTT(3 (4,5 dimethylthiazol 2 yl) 2,5 diphenytetrazolium bromide) assay, 3 H TdR incorporation, flow cytometry, fluorescence polarization assay, and soft agar culture. The results indicate that when the concentrations of LaCit are 0.05, 0.1, 0.5 and 1 mmol·L -1 , it inhibits PG cells proliferation. When the concentrations are 0.5 and 1 mmol·L -1 LaCit inhibits PG cells 3 H TdR incorporation, and decreases the proportion of PG cells in S phase while increases the proportion of PG cells in G 1 phase and decreases the membrane fluidity of PG cells. But LaCit has no effect on BGC 823 cells except for cell membrane fluidity and no effect on 2BS cells.展开更多
Various biological indices were used to observe the effects of Tween-80 in combination with hyperthermia at different temperature (39-43℃) for different period of time (20-100 minutes) on human stomach cell line BGC-...Various biological indices were used to observe the effects of Tween-80 in combination with hyperthermia at different temperature (39-43℃) for different period of time (20-100 minutes) on human stomach cell line BGC-823. The results showed that Tween-80obviously reduced the activation energy of BGC-823 cells. Synergistic effect was observed if applied with heat at 39℃ with the increase in temperature and time,the inhibitory effect on the cancer cells was gradually intensified. The lethal rate of BGC-823 cells treated by heat at 41℃ in combination with Tween-80 was around 5.2 times as that treated by hyperthermia alone. The synergistic effect of heat at 41℃ for 100 min. in combination with Tween-80 was equivalent to the effect of 43℃ for 100 min.In other words, the critical temperature for BGC-823 cells was hereby reduced about 2℃. The measurement of membrane mobility, SDH activity etc.also showed that at 41℃ the synergistic effect of hyperthermia in combination with Tween-80 was the best, it exceeding the single effect of these factors in combination and showing effect of multiplication. The synergistic effect of heat at 41℃ in combination with Tween-80 was higher than that of heat at 41℃ in combination with MMC. It also demonstrated that the specific and sustained action on the inhibition of cancer cells did exist. These studies suggested that the synergistic mechanism of Tween-80 and hyperthermia probably was that both of them acted on the cell membrane system. Supported by The National Natural Science Fund of China (No. 3860948)展开更多
文摘Objective:To explore the influence of silencing Bcl-2 expression by small interfering RNA（siRNA） on Bcl-2 protein expression,cell apoptosis rale and radiosensilivity of gastric cancer BCC823 cells.Methods:siRNA segment for Bcl-2 gene was designed and synthesized,then was induced into gastric cancer BGC 823 cells by liposome transfection.Bcl-2 protein expression was detected by Western Blotting.After X radiation,flow cytometry and clone forming assay were used to determine the effects of RNA interference on BGC823 cell apoptosis rate and radiosensitivity. Result:After the transfection of Bcl-2 siRNA,the positive expression rate of Bcl-2 protein in BGC823 cells was（35.45±2.35）%.Compared with the control group and negative siRNA transfection group,the rate was significantly decreased（P【0.01）.The apoptosis rate of BGC823-RNAi cell was（10.81±0.91）%,which was significantly higher than the control group and negative siRNA transfection group（P【0.01）.After 48h X radiation,the apoptosis rate of BGC823-RNAi was（28.91±1.40）%,which was significantly higher than the control group and the group without radiation （P【0.01）.During clone forming assay D<sub>0</sub>,D<sub>4</sub> and SF<sub>2</sub> values in Bcl-2 siRNA1 transfection group were all lower than those in the control group.The radiosensitivity ratio was 1.28（the ratio of D<sub>0</sub>） and 1.60（the ratio of D<sub>4</sub>）.Conclusions:Specific siRNA of Bcl-2 gene can effectively inhibit the expression of Bcl-2 gene,enhance the radiosensitivity and apoptosis of gastric cancer BGC823 cells,having good clinical application perspective.
基金Supported by National Key Technology Research and Development Program(2018FYC1708000)Basic Research Project for Application of Science and Technology in Sichuan Province(2017JY0274)Science and Technology Research Project of Sichuan Traditional Chinese Medicine Administration(2018JC028).
文摘[Objectives]This study aimed to investigate the inhibiting effect of Obazema on proliferation of gastric cancer BGC-823 cells and its mechanism.[Methods]BGC-823 cells were treated with high,medium and low concentrations of drug-containing serum(0.316%,0.158%and 0.079%)for 0,48,72 and 96 h,respectively.Then,the proliferation of the cells was detected with CCK-8 method,and the expression of related proteins,B lymphocytoma-2(Bcl-2),phosphorylated protein kinase B(p-Akt),protein kinase B(Akt)and glyceraldehyde-3-phosphate dehydrogenase(GAPDH),was detected using Western blotting.[Results]The proliferation of the BGC-823 cells was significantly inhibited with different doses of Boenninghausenia albiflora(Hook.)Reichb.ex Meisn.var.albiflora(CH)and B.sessilicarpa Lévl.(S)(P<0.01),in dose-dependent and time-dependent manners.The inhibition of high-dose S on cell proliferation was similar to that of CTX 48 h after administration;the inhibition of high-dose CH on cell proliferation was significantly stronger than that of CTX(P<0.01);different doses of drug administration groups significantly inhibited the expression of p-Akt and Bcl-2 in the BGC-823 cells;the inhibition of high-dose CH on the expression of P-Akt and Bcl-2 and the inhibition of medium-dose CH on the expression of Bcl-2 were significantly stronger than that of CTX(P<0.05,P<0.01),in a certain dose-dependent manner;at the same dose,the inhibition of CH on the expression of the proteins was stronger than that of S(P<0.05,P<0.01);administration of S and CH significantly inhibited the expression of GAPDH compared with CTX(P<0.05,P<0.01).[Conclusions]Obazema has the capacity to inhibit the proliferation of BGC-823 cells.The mechanism may be achieved by inhibiting the expression of p-Akt and Bcl-2,and GAPDH may be the target gene of its anti-tumor mechanism.The inhibiting effect of CH on BGC-823 cells was more significantly than that of S.
文摘The effect of LaCit on cell proliferation, DNA synthesize, cell cycle, cell membrane fluidity, growth in soft agar in human lung cancer cells PG and human gastric carcinoma cells BGC 823 was studied by MTT(3 (4,5 dimethylthiazol 2 yl) 2,5 diphenytetrazolium bromide) assay, 3 H TdR incorporation, flow cytometry, fluorescence polarization assay, and soft agar culture. The results indicate that when the concentrations of LaCit are 0.05, 0.1, 0.5 and 1 mmol·L -1 , it inhibits PG cells proliferation. When the concentrations are 0.5 and 1 mmol·L -1 LaCit inhibits PG cells 3 H TdR incorporation, and decreases the proportion of PG cells in S phase while increases the proportion of PG cells in G 1 phase and decreases the membrane fluidity of PG cells. But LaCit has no effect on BGC 823 cells except for cell membrane fluidity and no effect on 2BS cells.
文摘Various biological indices were used to observe the effects of Tween-80 in combination with hyperthermia at different temperature (39-43℃) for different period of time (20-100 minutes) on human stomach cell line BGC-823. The results showed that Tween-80obviously reduced the activation energy of BGC-823 cells. Synergistic effect was observed if applied with heat at 39℃ with the increase in temperature and time,the inhibitory effect on the cancer cells was gradually intensified. The lethal rate of BGC-823 cells treated by heat at 41℃ in combination with Tween-80 was around 5.2 times as that treated by hyperthermia alone. The synergistic effect of heat at 41℃ for 100 min. in combination with Tween-80 was equivalent to the effect of 43℃ for 100 min.In other words, the critical temperature for BGC-823 cells was hereby reduced about 2℃. The measurement of membrane mobility, SDH activity etc.also showed that at 41℃ the synergistic effect of hyperthermia in combination with Tween-80 was the best, it exceeding the single effect of these factors in combination and showing effect of multiplication. The synergistic effect of heat at 41℃ in combination with Tween-80 was higher than that of heat at 41℃ in combination with MMC. It also demonstrated that the specific and sustained action on the inhibition of cancer cells did exist. These studies suggested that the synergistic mechanism of Tween-80 and hyperthermia probably was that both of them acted on the cell membrane system. Supported by The National Natural Science Fund of China (No. 3860948)
