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1.
AIM: To investigate the effect on growth and activity of telomerase in esophageal carcinoma cells by inhibiting ubiquitin-proteasome pathway(UPP). METHODS: The esophageal carcinoma cell strain Eca9706 was treated with MG-132 to inhibit its UPP specially. The effect of growth suppression on cells was evaluated with MTT assay, morphologic changes of cells were observed under microscope, cell cycle and apoptosis were detected by flow cytometry (FCM). DNA fragment analysis was used to confirm the presence of apoptosis. The activity of telomerase was detected. RESULTS: MG-132 had obvious inhibitory effect on the growth of Eca9706 cells in a dose and time-dependent manner. Obvious pathologic change of cells were observed under microscope, cells became round, small and exfoliating. The FCM analysis showed that the ratio of esophageal carcinoma cells of G1 phase increased and a obviously apoptotic sub-G1 peak was found. Agarose electrophoresis showed marked ladder. The activity of telomerase was obviously inhibited. CONCLUSIONS: MG-132 significantly inhibits the growth and the activity of telomerase of Eca 9706 cells. These findings indicate that inhibiting UPP is a new strategy for the treatment of esophageal carcinoma.  相似文献   

2.
AIM:To study the mechanism of growth inhibitory effect of a selective cyclooxygenase-2 (COX-2) inhibitor,NS-398,on cancer cells.METHODS:The esophageal cancer cell line (EC9706),which expresses COX-2 constitutively,and hepatocellular carcinoma cell line (SMMC7721),which expresses no COX-2,were studied.The cell lines were incubated with NS-398 at doses of 10,20,50,100 μmol/L for 24 h,48 h and 72 h.Antiproliferation effect was measured by ［3H］-TdR incorporation.The cell apoptosis were determined by flow cytometry (FCM) and DNA fragmentation analysis.Survivin was detected by immunocytochemical technique.RESULTS:The growth inhibition was induced by NS398 in a dose- and time-dependent manners in both cell lines.FCM analysis revealed a high sub-G1 cell peak in EC9706 group and agarose electrophoresis showed marked apoptosis ladder pattern.However,no apoptosis was observed in SMMC7721 cells treated with NS-398.The difference of apoptosis percentage in EC9706 and SMMC7721 was (45.23±1.08)% and (3.05±0.15)% (P<0.01).After 24 h incubation with NS-398 at concentration of 100 μmol/L,the expression of survivin was markedly reduced in EC9706,no change was observed in SMMC7721.CONCLUSION:NS-398 suppresses cell growth in cancer cell lines by different mechanism.NS-398 suppresses cell growth and increases apoptosis in the cancer cells that expresses COX-2.  相似文献   

3.
AIM: To observe the influence of human mutant p27 gene (p27mt) on the growth and so as to investigate the function and mechanism of p27mt in gene therapy for colorectal cancer.METHODS: Colorectal cancer cell line SW480 was infected with recombinant replication defective adenovirus Ad-p27mt,and expression of p27mt protein was detected by Western blotting.The inhibitory effect of p27mt on SW480 and cell cycle were determined by flow cytometry,and DNA fragment was analyzed to identify the occurrence of apoptosis.RESULTS: After transfected with Ad-p27mt,p27 protein was highly expressed in SW480 cells.77.96% colorectal cancer cells were blocked in phase G0/G1,while in Ad-LacZ group and blank control group,27.57% and 25.29% cells were blocked in the same phase,respectively.Growth curve showed Ad-p27mt had an obviously inhibitory effect on the growth of SW480 cells.DNA fragment assay demonstrated that p27mt was able to induce the apoptosis of colorectal cancer cells.CONCLUSION: p27mt has an obvious blocking effect on colorectal cancer cell cycle,and most cells are blocked in phase G0/G1.This blockage is related with the growth inhibition and apoptosis induced by p27mt.  相似文献   

4.
AIM:To observe the effect of simvastatin on the proliferation of vascular smooth muscle cells(VSMCs) induced by serum and growth factor PDGF-BB and the effect of simvastatin on the expression of PTEN,a important regulator of G1/S cell cycle transition. METHODS:The DNA synthesis was determined by [3H]-TdR incorporation, cell cycle was examined with flow cytometry, the protein level of PTEN was measured by Western blot method. RESULTS: (1)Simvastatin inhibited [3H]-TdR incorporation in a dose dependent manner. (2) Flow cytometric DNA analysis revealed that simvastatin induced significantly enhancement of G0/G1 phase and decrease in S phase VSMCs.(3)Simvastatin increased protein level of PTEN and mevalonate, a metabolite of HMG-COA, reversed the effect of simvastatin on PTEN protein expression. CONCLUSION:Simvastatin may inhibit proliferation of VSMCs and retarded cell cycle in G0/G1 phase by increasing PTEN expression through inhibiting synthesis of mevalonate.  相似文献   

5.
AIM: To investigate the effect of probucol on proliferation of rat vascular smooth muscle cells(VSMC) stimulated by basic fibroblast growth factor (bFGF) and/or hydrogen peroxide(H2O2). METHODS: Effects of probucol on VSMC proliferation and DNA synthesis stimulated by bFGF and/or H2O2 were observed by means of MTT test, cell number count and [3H]-TdR incorporation. RESULTS: ①Probucol significantly inhibited proliferation and DNA synthesis in VSMC stimulated by bFGF and/or H2O2, with dosage-dependent manner. Cell number, A value and [3H]-TdR incorporation in group probucol+bFGF and group probucol+H2O2 were reduced by 40.0%, 39.1%, 45.5% and 46.9%, 45.0%, 39.5%, respectively, compared with group bFGF and group H2O2 (P＜0.05, P＜0.01, respectively). ②Pretreatment of VSMC with probucol for 24 h prior to bFGF and/or H2O2 stimulation exhibited significant inhibiton of VSMC proliferation and DNA synthesis, but after prestimulation by bFGF and/or H2O2 for 24 h, probucol had no influence on VSMC proliferation and DNA synthesis (P＞0.05). CONCLUSION: Probucol dramatically inhibited proliferation and DNA synthesis in VSMC stimulated by bFGF and/or H2O2, but had no inhibitory effect on the cell proliferation prestimulated by bFGF and /or H2O2.  相似文献   

6.
AIM: To investigate the change of chemosensitivity of esophageal carcinoma cells before and after induction by radiation,and to detect the excision repair cross complementation group 1 (ERCC1) gene before and after induction,and further to analyze the relationship between ERCC1 expression and chemosensitivity.METHODS: The esophageal carcinoma cell EC9706 was radiated repeatedly by a long-term,intermittent ［60Co］-γ radiation to induce the radioresistance esophageal carcinoma cell EC9706-R.The IC50 and resistant index (RI) of EC9706 and EC9706-R were detected by MTT assay.The expression of ERCC1 was examined by immunocytochemistry and RT-PCR.RESULTS: The IC50 of EC9706 and EC9706-R cells to cisplatin were (1.480±0.012) mg/L and (1.836±0.008) mg/L,respectively (P<0.05),and the RI was 1.240±0.015.The tinctorial scores of ERCC1 protein in EC9706 and EC9706-R were 2.838±0.055 and 2.898±0.039 (P>0.05),and the relatively quantities (IDV) of ERCC1 mRNA in EC9706 and EC9706-R cells were 1.168±0.068 and 1.143±0.089 (P>0.05).SP and RT-PCR did not display visible difference in protein level and mRNA level.CONCLUSION: The chemosensitivity of EC9706-R cells to cisplatin is decreased compared with EC9706 cells,but ERCC1 expression does not change with the radioresistance and chemoresistance.  相似文献   

7.
AIM: To investigate the effects of platelet-derived growth factor on DNA and collagen protein synthesis in human vascular fibroblasts. METHODS: In the present experiment, the human vascular fibroblasts were cultured and effects of platelet-derived growth factor-BB on DNA and collagen protein synthesis in human vascular fibroblasts were observed by using [3H]-TdR incorporation and [3H]-proline incorporation in vitro. RESULTS: Platelet-derived growth factor-BB significantly promoted NDA synthesis and collagen protein synthesis of quiescent human vascular fibroblasts, with a maximal response at a concentration of 30μg·L-1at 24 h and 36 h, respectively. CONCLUSION: Platelet-derived growth factor-BB promotes DNA and collagen protein synthesis in cultured human vascular fibroblasts.  相似文献   

8.
AIM:To explore the influence of ginsenoside Rh2 on cultured PC-3M cell cycle in vitro. METHODS:DNA content was measured using [3H]-TdR incorporation assay. To analysis the cycle changes of PC-3M cells, flow cytometry was used in the experiment.RESULTS:The results indicated that the rate of [3H]-TdR incorporation was lower in Rh2-treated cells than the control in a dose-dependent manner. Flow cytometry demonstrated that the pecent of G1 phase treated with Rh2 was higher than that of control. CONCLUSION:These results suggested that PC-3M cells cultured with Rh2 for 24 h were blocked at G1 phase, and Rh2 inhibited the growth of PC-3M cells in vitro.  相似文献   

9.
AIM: To investigate the effect of Salidroside on the proliferation, DNA synthesis, intracellular Ca2+ content of rabbit PASMC (pulmonary artery smooth muscle cells) under hypoxia. METHODS: Techniques of cell culture, MTT test, [3H][3H][3H]-TdR incorporation, fluo-3 and confocal laser scanning microscopy were used. RESULTS: The A value of MTT and [3H][3H]-TdR incorporation of PASMC increased significantly by 62% (P＜0.05) and 138% (P＜0.01) after 24 h hypoxia. Salidroside (32×10-5 mol/L) inhibited the action of hypoxia on the proliferation of PASMC, the A value of MTT and [3H][3H]-TdR incorporation declined significantly by 29% (P＜0.05) and 37% (P＜0.01) compared with hypoxia group. A calcium channel blocker, verapamil could also inhibit the accelerative effect of hypoxia on the proliferation of PASMC. The intracelluler Ca2+ content of PASMC raised markedly under hypoxia, but the effect of hypoxia on the intracelluler Ca2+ content could be inhibited by Salidroside. CONCLUSION: Salidroside inhibited the proliferation, DNA synthesis of PASMC induced by hypoxia. The inhibitory action of Salidroside on the increase in intracellular Ca2+ concentration under hypoxia might be one of the mechenisms.  相似文献   

10.
AIM: To investigate the effects of intracellular free calcium ([Ca2+]i) from different resources on the proliferation mediated by mitogen activated protein kinase (MAPK) in vascular smooth muscle cells (VSMCs). METHODS: Cultured VSMCs were used in all experiments. Calcium influx was stimulated by angiotension Ⅱ(Ang Ⅱ). The release of intracellular calcium stores was induced by inositol trisphosphate (IP3) and ryanodine (RY). MAPK activity was measured by [γ-32P]-ATP incorporation MAPK protein expression by western blot, VSMCs proliferation by [3H]-Leucine ([3H]-Leu) and [3H]-Thymidine ([3H]-TdR) incorporation. RESULTS: Compared to the control VSMCs, Ang Ⅱ, IP3 and RY significantly increased [Ca2+]i concentration activity of MAPK and its protein content in VSMCs. The promotion of [3H]-Leu and [3H]-TdR incorporation in VSMCs was also observed (P<0.01). CONCLUSION: The study indicated that calcium activator-induced increase in the activity and protein content of MAPK was involved in the proliferation of VSMCs, which was closely related to the [Ca2+]i concentration but independent to its origin.  相似文献   

11.
AIM: To investigate the effect of microRNA-24-3p (miR-24-3p) on the viability and apoptosis of esophageal cancer cells. METHODS: The expression of miR-24-3p and KLF6 mRNA in the esophageal cancer cells TE11, Eca109 and EC9706 were detected by RT-qPCR. The protein expression of KLF6 was determined by Western blot. EC9706 cells were transfected with anti-miR-24-3p and KLF6 siRNA. The cell viability was measured by MTT assay, the apoptotic rate was analyzed by flow cytometry, and the proliferation, apoptosis and IL-6/STAT3 signaling pathways related proteins were determined by Western blot. The level of IL-6 was measured by ELISA. The dual luciferase reporter gene assay was used to verify the relationship between miR-24-3p and KLF6. RESULTS: The levels of miR-24-3p were up-regulated in the esophageal cancer cells TE11, Eca109 and EC9706 (P < 0.05), and the expression of KLF6 at mRNA and protein levels was down-regulated (P < 0.05). Knock-down of miR-24-3p expression inhibited the cell viability, induced apoptosis, and inhibited the protein levels of CDK4, cyclin D1, CDC25A, p-STAT3, Bcl-2 and IL-6, and promoted the protein expression of caspase-3 and Bax in EC9706 cells. CONCLUSION: miR-24-3p targets KLF6 gene to affect the viability and apoptosis of esophageal cancer cells by regulating IL-6/STAT3 signaling pathway.  相似文献   

12.
AIM: To investigate the effect of taurine on calcification of vascular smooth muscle cells (VSMCs).METHODS: Calcified VSMCs of rat in vitro were induced by β-glycerophosphate. Cellular calcium content, alkaline phosphatase(ALP) activities and [45Ca]accumulation were measured. DNA synthesis were evaluated by [3H]-thymidine ( [3H]-TdR) incorporation. RESULTS: Calcium content, ALP activities and [45Ca]uptake of calcified VSMCs stimulated by taurine (5-20 mmol/L) were greatly decreased in a concentration-dependent manner as compared with calcified group (P＜0.01). Taurine also inhibited the proliferation of calcified cells in a concentration-dependent manner. Cell countingz, [3H]-TdR incorporation of calcified cells stimulated by taurine were greatly decreased as compared with calcified VSMCs (P＜0.01). CONCLUSION: It was demonstrated that calcification of VSMCs may be alleviated by taurine.  相似文献   

13.
AIM: To study the effect of homocysteine (HCY) on proliferation of airway smooth muscle cells and fibroblasts and the effect of HCY on collagen prodution of airway fibroblasts. METHODS: [3H]-TdR incorpora- tion was measured in cultured airway smooth muscle cells. The [3H]-TdR and [3H]-proline incorporation were mea- sured in cultured airway fibroblasts. RESULTS: HCY induced proliferation of airway smooth muscle cells and fibroblasts in a concentration - dependent manner. HCY also induced collagen production of airway fibroblasts in a concentration - dependent manner. The inhibitors of protein kinase C, H7 and polymyxin B, inhibited HCY - induced proliferation of airway smooth muscle cells. CONCLUSIONS: HCY induced proliferation of airway smooth muscle cells and fibroblasts, HCY also induced collagen production of airway fibroblasts. The HCY - induced proliferation of airway smooth muscle cells may be related to the pathway of PKC signal transduction.  相似文献   

14.
AIM: To investigate the influence of DNA polymerase β(DNA polβ) on the biological characteristics of esophageal carcinoma cells.METHODS: siRNA expression plasmid targeting DNA polβ was constructed and transfected into EC9706 cells. The positive colonies were screened by G418 selection. The proliferation of EC9706 cells was detected by flow cytometry and nude mice xenograft model. The expression of DNA polβ was detected by real-time PCR. RESULTS: Compared to empty vector group and control group, the expression of DNA polβ was significantly reduced in the cells transfected with siRNA expression plasmid pSINsi-hU6-siRNA polβ288(experimental group 1) and the cells transfected with siRNA expression plasmid pSINsi-hU6-siRNA polβ814(experimental group 2). The expression of DNA polβ was significantly increased in DNA polβ over-expression group(P＜0.05). The proportion of the cells in S phase increased significantly in experimental group 1, experimental group 2 and DNA polβ over-expression group(P＜0.05). The weight of tumors increased significantly(P＜0.05).CONCLUSION: The over-expression and extremely low expression of DNA polβ may increase genetic instability of the cells and play a role in the occurrence of esophageal carcinoma.  相似文献   

15.
AIM: To study the inducing effect of human mutant p27 gene on apoptosis of the colorectal cancer cells. METHODS: The recombinant adenovirus Ad-p27mt was constructed to infect the colorectal cancer cell SW480. The inducing effect of Ad-p27mt on apoptosis in colorectal cancer cells was measured by flow cytometry, DNA fragment analysis and TUNEL method. RESULTS: Ad-p27mt was successfully constructed. When the multiplicity of infection (MOI) was ≥50, the infection efficiency reached 100%. After 24 h of infection, there was an apoptotic hypodiploid peak observed by flow cytometry before G1 and there were apoptotic characteristic bands in the DNA electrophoresis. The apoptotic index detected by TUNEL method was 82.6±3.2 (Ad-p27mt group) and 5.0±3.5 (control group), respectively, the difference of which was significant (P<0.01). CONCLUSION: Human mutant p27 gene transfection effectively induces apoptosis in the colorectal cancer cells.  相似文献   

16.
Abstract: AIM: To investigate the effects of esophageal cancer-related gene 2 (ECRG2) protein combined with cisplatin (DDP) on the proliferation and apoptosis of human esophageal cancer EC9706 cells. METHODS: Methylthiazolyl tetrazolium (MTT) assay was used to examine the effects of single ECRG2 protein and ECRG2 protein combined with DDP on the proliferation of EC9706 cells. Hoechest 33258 staining was applied to analyze the effect of single ECRG2 protein and ECRG2 protein combined with DDP on apoptosis of EC9706 cells. The expression of p53 in EC9706 cells was detected by Western blotting. RESULTS: ECRG2 protein inhibited the proliferation of EC9706 cells. ECRG2 protein combined with DDP enhanced the inhibitory effect time- and dose-dependently in a certain range of concentrations. The number of apoptotic cells after treated with ECRG2 protein combined with DDP for 24 h was larger than those treated with single ECRG2 protein. The proliferation-inhibitory and qpoptosis-inducing expression of p53 was up-regulated in ECRG2 protein and DDP combination group compared with single ECRG2 protein group. CONCLUSION: DDP enhances the proliferation-inhibitory and apoptosis-inducing effects of ECRG2 protein on EC9706 cells. The apoptosis-inducing mechanism may be related to the up-regulation of p53 expression.  相似文献   

17.
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18.
CAI Wei  XU Yi 《园艺学报》2004,20(9):1627-1630
AIM: To observe the effects of Yangxue qingnao-containing serum on rat vascular smooth muscle cell (VSMC) proliferation induced by lysophosphatidic acid (LPA). METHODS: The [3H]-TdR incorporation and mitogen-activated protein kinasc (MAPK) activity were measured in cultured VSMC. End product of lipid peroxidation-MDA levels were also detected. RESULTS: 1×10－9,1×10－8 and 1×10－7 mol/L LPA enhanced the cultured VSMC [3H]-TdR incorporation, increased MAPK activity and MDA content in a concentration-dependent manner. 5％, 10％ and 15％ Yangxue qingnao-containing serum concentration-dependently inhibited the increase in VSMC [3H]-TdR incorporation, MAPK activity and MDA content induced by LPA. CONCLUSIONS: LPA has a stimulating effect on VSMC proliferation. The LPA-induced intracellular signal transduction may be related to MAPK activity. Yangxue-qingnao can efficiently inhibit LPA -induced VSMC proliferation,MAPK activity and lipid peroxidation.  相似文献   

19.
AIM:To investigate the effects of fibronectin (FN) on the proliferation and collagen synthesis in cultured rat cardiac fibroblasts (CFb) derived from SHR (CFbSHR) and WKY (CFbWKY). METHODS:CFb derived from 12-week-old spontaneously hypertensive rat (SHR) and WKY was cultured by outgrowth of tissue block. Cell proliferation of CFb was measured by cell number counting and[3H]-TdR incorporation using 24-well plates pre-coated with 5 μg/cm2 of FN. Collagen synthesis was determined by [3H]-proline incorporation. RESULTS:As compared with control, the cell number of fibroblasts derived from SHR and WKY were significantly increased to 163.75% and 170.42% respectively after 72 h incubation with FN in the presence of 0.4% FCS from a intial cell density of 1×104 cells/mL. DNA synthesis of CFb was markedly promoted by FN. FN induced an increased in [3H]-proline incorporation in both CFbSHR and CFbWKY. CONCLUSION:FN is able to promote cell proliferation and collagen synthesis of CFb derived both from SHR and WKY.  相似文献   

20.
番茄E8启动子乙烯应答元件克隆及DNA序列分析   总被引：2，自引：0，他引：2  
E8启动子是常用的番茄果实特异表达启动子之一，是指番茄E8基因5’侧翼近2．2kb的DNA序列。前人的研究表明，E8基因5’侧翼-2181～-1088区段的删除使E8基因表达量大幅度下降(仅为完整E8启动子的1／10)，同时该区段还是E8基因对乙烯应答的充分必要区域；这说明了该区段是E8基因表达调控的重要元件。  相似文献   

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