AIM: Heat shock protein (HSP)70 is over-expressed in human gastric cancer and plays an important role in the progression of this cancer. We investigated the effects of antisense HSP70 oligomer on human gastric cancer ...AIM: Heat shock protein (HSP)70 is over-expressed in human gastric cancer and plays an important role in the progression of this cancer. We investigated the effects of antisense HSP70 oligomer on human gastric cancer cell line SGC-7901, and its potential role in gene therapy for this cancer.METHODS: Human gastric cancer cell line SGC-7901 was treated in vitro with various concentrations of antisense HSP70 oligonucleotides at different intervals. Growth inhibition was determined as percentage by trypan blue dye exclusion test. Extracted DNA was electrophoresed on agarose gel, and distribution of cell cycle and kinetics of apoptosis induction were analyzed by propidium iodide DNA incorporation using flow cytometry, which was also used to detect the effects of antisense oligomer pretreatment on the subsequent apoptosis induced by heat shock in SGC-7901 cells. Proteins were extracted for simultaneous measurement of HSP70 expression level by SDS-PAGE Western blotting.RESULTS: The number of viable cells decreased in a doseand time-dependent manner, and ladder-like patterns of DNA fragments were observed in SGC-7901 cells treated with antisense HSP70 oligomers at a concentration of 10 μmol/L for 48 h or 8 μmol/L for 72 h, which were consistent with inter-nucleosomal DNA fragmentation. Flow cytometric analysis showed a dose- and time-dependent increase in apoptotic rate by HSP70 antisense oligomers. This response was accompanied with a decrease in the percentage of cells in the G1 and S phases of the cell cycle, suggesting inhibition of cell proliferation. In addition, flow cytometry also showed that pretreatment of SGC-7901 cells with HSP70 antisense oligomers enhanced the subsequent apoptosis induced by heat shock treatment. Western blotting demonstrated that HSP70 antisense oligomers inhibited HSP70 expression, which preceded apoptosis, and HSP70 was undetectable at the concentration of 10 μmol/L for 48 h or 8 μmol/L for 72 h.CONCLUSION: Antisense HSP70 oligomers can abrogate HSP70 expression in SGC-7901 cells, wh展开更多
Objective To isolate and identify the bioactive phytochemicals from the leaves of Camellia nitidissima. Methods The chemical constituents were isolated and purified by repeated silica gel, Sephadex LH-20, MCI gel colu...Objective To isolate and identify the bioactive phytochemicals from the leaves of Camellia nitidissima. Methods The chemical constituents were isolated and purified by repeated silica gel, Sephadex LH-20, MCI gel columns, recrystallization, and semi-preparative HPLC techniques. The chemicl structures of these compounds were identified on the basis of spectral data including NMR and MS. Then quorum sensing inhibition （QSI） activities of these compounds were tested using Chromobacterium violaceum CV026 as the bioindicator strain. The antitumor activities of these compounds were measured using SGC7901 as cell proliferation and cytotoxicity. Results cx-Spinasteryl-I^-D-glucopyranoside （1）, stigmasta-7,22-diene-3-O-[c^-L-arabinopyranosyl （1 -2）]-β-D-galactopyranoside （2）, kaempferol 3-O-[2-O-（trans-p-coumaroyl）-3-O-α -D-glucopyranosyl]-α-D-glucopyranoside （3）, aromadendrin （4）, catechin （5）, phlorizin 4＇-O-β-D-glucopyranoside （6）, （3R,6R,7Lg-3-hydroxy-4,7-megastigmadien- 9-one （7）, dodecanoic acid （8）, 3α-acetoxy-20-1upanol （9）, and 3β,6α- trihydroxyolean- 7-one （1 0） were successively isolated from the leaves of C. nitidissima. Unfortunately, these compounds had no QSI activity. Based on Cell Counting Kit-8 （CCK-8） assay, compound 10 showed the best anti-tumor activity or all compounds （ICs0 = 91.7 μg/mL）. Conclusion Apart from compounds 4 and 5, other eight compounds are reported in this plant for the first time. All compounds show no QSI activity, compound 10 shows potential cytotoxic activity on SGC7901 cells in vitro.展开更多
AIM: To determine the effect of apoptosis on gastric cancer cells (SGC-7901) induced by cis-9, trans-11-conjugated linoleic acid (c9, t11-CLA) and its possible mechanism in the inhibition of cancer cells growth.METHOD...AIM: To determine the effect of apoptosis on gastric cancer cells (SGC-7901) induced by cis-9, trans-11-conjugated linoleic acid (c9, t11-CLA) and its possible mechanism in the inhibition of cancer cells growth.METHODS: Using cell culture, flow cytometery and immunocytochemical techniques, we examined the cell growth, frequency of apoptosis and distribution of cell cycle,expression of ki67, bcl-2, Fas, and c-myc of SGC-7901 cells which were treated with various c9, t11-CLA concentrations (25,50,100 and 200 μmol@L-1) of c9, t11-CLA for 24h and 48 h,with a negative control (0.1% ethanol).RESULTS: The growth of SGC-7901 cells was inhibited by c9,t11-CLA. Eight days after treatment with various concentrations of c9,t11-CLA, as mentioned above, the inhibition rates were 5.9 %, 20.2 %,75.6 % and 82.4 %, respectively. The frequency of apoptosis on SGC-7901 cells induced by different concentrations of c9, t11-CLA (except for 25 μmol@L-1, 24 h) was significantly greater than that in the negative control (P＜0.01). To further investigate the influence of the cell cycle progression, we found that apoptosis induced by c9, t11-CLA may be involved in blocking the cell cycle of SGC-7901 cells. Immunocytochemical staining demonstrated that SGC-7901 cells preincubated in media supplemented with different c9, t11-CLA concentrations for various time periods significantly decreased the expressions of ki67 (the expression rates were 18.70-3.20 %, at 24 h and 8.10-0.20 % at 48 h, respectively), bd-2 (4.30-0.15 % at 24 h and 8.05 %-0 at 48 h),and c-myc(4.85-2.20 % at 24 h and 4.75-0.30 % at 48 h) as compared with those in the controls (the expressions of ki67, bcl-2, and c-mycwere 15.1% at 24 h and 13.5 % at 48 h, 6.80 % at 24 h and 8.00 % at 48 h,5.50 % at 24 h and 5.30 % at 48 h, respectively) (P＜0.01),whereas the expressions of Fas were increased (0.60-2.75 %,24 h and 0.45-5.95 %, 48 h).CONCLUSION: The growth and proliferation of SGC-7901 cells are inhibited by cg, t11-CLA via blocking the cell cycle,pathways of bcl-2-associate展开更多
文摘AIM: Heat shock protein (HSP)70 is over-expressed in human gastric cancer and plays an important role in the progression of this cancer. We investigated the effects of antisense HSP70 oligomer on human gastric cancer cell line SGC-7901, and its potential role in gene therapy for this cancer.METHODS: Human gastric cancer cell line SGC-7901 was treated in vitro with various concentrations of antisense HSP70 oligonucleotides at different intervals. Growth inhibition was determined as percentage by trypan blue dye exclusion test. Extracted DNA was electrophoresed on agarose gel, and distribution of cell cycle and kinetics of apoptosis induction were analyzed by propidium iodide DNA incorporation using flow cytometry, which was also used to detect the effects of antisense oligomer pretreatment on the subsequent apoptosis induced by heat shock in SGC-7901 cells. Proteins were extracted for simultaneous measurement of HSP70 expression level by SDS-PAGE Western blotting.RESULTS: The number of viable cells decreased in a doseand time-dependent manner, and ladder-like patterns of DNA fragments were observed in SGC-7901 cells treated with antisense HSP70 oligomers at a concentration of 10 μmol/L for 48 h or 8 μmol/L for 72 h, which were consistent with inter-nucleosomal DNA fragmentation. Flow cytometric analysis showed a dose- and time-dependent increase in apoptotic rate by HSP70 antisense oligomers. This response was accompanied with a decrease in the percentage of cells in the G1 and S phases of the cell cycle, suggesting inhibition of cell proliferation. In addition, flow cytometry also showed that pretreatment of SGC-7901 cells with HSP70 antisense oligomers enhanced the subsequent apoptosis induced by heat shock treatment. Western blotting demonstrated that HSP70 antisense oligomers inhibited HSP70 expression, which preceded apoptosis, and HSP70 was undetectable at the concentration of 10 μmol/L for 48 h or 8 μmol/L for 72 h.CONCLUSION: Antisense HSP70 oligomers can abrogate HSP70 expression in SGC-7901 cells, wh
基金National High Technology Research and Development Program of China(863 Program)(2014AA022208)National Natural Science Foundation of China(31170131 and 31070312)Jiangsu Qinglan Project
文摘Objective To isolate and identify the bioactive phytochemicals from the leaves of Camellia nitidissima. Methods The chemical constituents were isolated and purified by repeated silica gel, Sephadex LH-20, MCI gel columns, recrystallization, and semi-preparative HPLC techniques. The chemicl structures of these compounds were identified on the basis of spectral data including NMR and MS. Then quorum sensing inhibition （QSI） activities of these compounds were tested using Chromobacterium violaceum CV026 as the bioindicator strain. The antitumor activities of these compounds were measured using SGC7901 as cell proliferation and cytotoxicity. Results cx-Spinasteryl-I^-D-glucopyranoside （1）, stigmasta-7,22-diene-3-O-[c^-L-arabinopyranosyl （1 -2）]-β-D-galactopyranoside （2）, kaempferol 3-O-[2-O-（trans-p-coumaroyl）-3-O-α -D-glucopyranosyl]-α-D-glucopyranoside （3）, aromadendrin （4）, catechin （5）, phlorizin 4＇-O-β-D-glucopyranoside （6）, （3R,6R,7Lg-3-hydroxy-4,7-megastigmadien- 9-one （7）, dodecanoic acid （8）, 3α-acetoxy-20-1upanol （9）, and 3β,6α- trihydroxyolean- 7-one （1 0） were successively isolated from the leaves of C. nitidissima. Unfortunately, these compounds had no QSI activity. Based on Cell Counting Kit-8 （CCK-8） assay, compound 10 showed the best anti-tumor activity or all compounds （ICs0 = 91.7 μg/mL）. Conclusion Apart from compounds 4 and 5, other eight compounds are reported in this plant for the first time. All compounds show no QSI activity, compound 10 shows potential cytotoxic activity on SGC7901 cells in vitro.
基金the National Natural Science Foundation of China,No.39870661
文摘AIM: To determine the effect of apoptosis on gastric cancer cells (SGC-7901) induced by cis-9, trans-11-conjugated linoleic acid (c9, t11-CLA) and its possible mechanism in the inhibition of cancer cells growth.METHODS: Using cell culture, flow cytometery and immunocytochemical techniques, we examined the cell growth, frequency of apoptosis and distribution of cell cycle,expression of ki67, bcl-2, Fas, and c-myc of SGC-7901 cells which were treated with various c9, t11-CLA concentrations (25,50,100 and 200 μmol@L-1) of c9, t11-CLA for 24h and 48 h,with a negative control (0.1% ethanol).RESULTS: The growth of SGC-7901 cells was inhibited by c9,t11-CLA. Eight days after treatment with various concentrations of c9,t11-CLA, as mentioned above, the inhibition rates were 5.9 %, 20.2 %,75.6 % and 82.4 %, respectively. The frequency of apoptosis on SGC-7901 cells induced by different concentrations of c9, t11-CLA (except for 25 μmol@L-1, 24 h) was significantly greater than that in the negative control (P＜0.01). To further investigate the influence of the cell cycle progression, we found that apoptosis induced by c9, t11-CLA may be involved in blocking the cell cycle of SGC-7901 cells. Immunocytochemical staining demonstrated that SGC-7901 cells preincubated in media supplemented with different c9, t11-CLA concentrations for various time periods significantly decreased the expressions of ki67 (the expression rates were 18.70-3.20 %, at 24 h and 8.10-0.20 % at 48 h, respectively), bd-2 (4.30-0.15 % at 24 h and 8.05 %-0 at 48 h),and c-myc(4.85-2.20 % at 24 h and 4.75-0.30 % at 48 h) as compared with those in the controls (the expressions of ki67, bcl-2, and c-mycwere 15.1% at 24 h and 13.5 % at 48 h, 6.80 % at 24 h and 8.00 % at 48 h,5.50 % at 24 h and 5.30 % at 48 h, respectively) (P＜0.01),whereas the expressions of Fas were increased (0.60-2.75 %,24 h and 0.45-5.95 %, 48 h).CONCLUSION: The growth and proliferation of SGC-7901 cells are inhibited by cg, t11-CLA via blocking the cell cycle,pathways of bcl-2-associate
