共查询到20条相似文献,搜索用时 0 毫秒
1.
WANG Hua ZHOU Bin GUO Xing ZHU Jun-ya CHENG Chun ZHU Chang-lai ZHANG Tian-yi 《园艺学报》2011,27(6):1226-1229
AIM: To investigate the changes of cell growth and cytoskeleton in hepatocarcinoma SMMC-7721 cells treated with ginsenoside Rh2.METHODS: Cell viability and apoptosis under the conditions of ginsenoside Rh2 exposure at different concentrations were measured by MTT test and flow cytometry,respectively. The morphological changes of F-actin labeled with FITC-phalloidin were observed under confocal laser scanning microscope. The structures of nuclear matrix-intermediate fibre system were observed under transmission electron microscope (TEM).RESULTS: Rh2 at 40 mg/L for 4 days inhibited the proliferation and induced apoptosis in SMMC-7721 cells more than those in control group, 10 mg/L Rh2 group and 20 mg/L Rh2 group. The F-actin in the cells treated with Rh2 was well-distributed, lined up in order and the number of fibers increased, while those in the control cells were in disorder and punctiform. The results of whole mount TEM indicated that the intermediate fiber was plentiful, well-distributed and interweaved into a regular network in Rh2 treated cells.CONCLUSION: Rh2 effectively inhibits the cell proliferation, increases the cell apoptosis and induces the change of the cytoskeleton alignment in SMMC-7721 cells. 相似文献
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AIM:To examine the effects of recombinant human hepatocyte growth factor(rhHGF) and native calf HGF(cHGF) on SMMC-7721 human hepatocellular carcinoma(HCC) cell line. METHODS:Human HCC cell line culture, photometric assay, and flow cytometric assay were used in this study .RESULTS:A similar type of dose-dependent cell growth inhibition effect on SMMC-7721 human HCC cells by rhHGF(5-20 μg/L) as well as by cHGF(25-100 mg/L) had been found, with the maximal effect at the highest concentration used. Approximately over 50% of the cells treated with rhHGF(5 μg/L, 10 μg/L, 20 μg/L) accumulated in the quiescent G0/G1 phase of the cell cycle over incubation periods for 3 d. CONCLUSION:The growth of SMMC-7721 human HCC cells was strongly inhibited by both rhHGF and cHGF. This might be because the cells exposed to HGF became arrested in the G0/G1 phase. 相似文献
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AIM: To investigate the effect of platelet-derived growth factor (PDGF) on β3 integrin gene expression and the role of β3 integrin on adhesion, migration and proliferation of vascular smooth muscle cells (VSMC) induced by PDGF. METHODS: β3 integxin gene expression was detected by RT-PCr. After β3 integrin extracellular do-main was blocks, VSMC adhesio, migration and proliferation were measured by adhesion assay awound-culture model an [3H]-TSR incorporation respectively.RESULTS: After the interaction between β3 integrin and extracellular matrix was blocked, VSMC proliferation was inhibited in some degree and the rate of [3H]-TdR incorporation into VSMC decreased 39%. The cell adhesion and migration were significantly inhibited when 10 mg/L anti-β3 integrin antibody was added (P<0.05). When VSMC were treated by PDGF for 6 hours, the expression of β3 integrin gene was 87% higher than that of control. CONCLUSION: PDGF significantly induces expression of β3 integrin gene in VSMC, and the interaction between β3 integrin and ECM protein may play an important role in VSMC adhesion and migration. 相似文献
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AIM: To explore the different inhibitory effect of arsenic trioxide (As2O3) on hepatocarcinoma cell growth in SMMC-7721 and BEL-7402 cell lines and its mechanism. METHODS: The cell culture and trypan blue staining were used to study the inhibitory effect of arsenic trioxide on cell growth, and the glutathione (GSH) contents in hepatocarcinoma cells treated with arsenic trioxide were detected. RESULTS: Arsenic trioxide inhibited the growth of BEL-7402 cells in a time and dose-dependent manner. The inhibitory effect was significant at a lower dose of 0.50 μmol/L for 24 h, however, to SMMC-7721 cells, a higher dose of 2.00 μmol/L for 96 h was needed. The inhibitory rate of arsenic trioxide (0.25-2.00 μmol/L) on BEL-7402 cell growth was higher than that on SMMC-7721 cells. The content of GSH in SMMC-7721 cells was much higher than that in BEL-7402 cells . CONCLUSION: There was a significant difference in inhibition of hepatocarcinoma cell growth by arsenic trioxide between BEL-7402 and SMMC-7721 cell lines, the cause of which may be due to the difference in GSH content in BEL-7402 and SMMC-7721 cells. 相似文献
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AIM: To explore the effects of kaempferol on the proliferation, invasion and migration abilities of HBx-HepG2 cells and to examine the underlying molecular mechanisms. METHODS: The expression levels of related genes at mRNA and protein levels were determined by RT-qPCR and Western blot. The cell apoptotic rate was analyzed by flow cytometry. The cell proliferation, growth, invasion and migration abilities were measured by MTT assay, colony formation assay, Transwell invasion assay and wound healing assay, respectively. RESULTS: Kaemferol inhibited HBx-HepG2 cell proliferation in a concentration-and time-dependent manner. Kaempferol at 100 μmol/L significantly inhibited the colony formation, invasion and migration abilities of the HBx-HepG2 cells. Kaemferol at 100 μmol/L also increased cell apoptotic rate, increased the protein levels of cleaved caspase-3, cleaved caspase-9 and Bax, and decreased the expression level of Bcl-2. In addition, kaemferol at 100 μmol/L suppressed the mRNA and protein expression levels of β-catenin, c-Myc and cyclin D1 in the HBx-HepG2 cells. Kaemferol at 100 μmol/L also suppressed the protein level of p-GSK-3β and the β-catenin protein levels in both cytoplasm and nucleus. LiCl treatment reversed the inhibitory effect of kaempferol on the growth, invasion and migration of the HBx-HepG2 cells. CONCLUSION: Kaempferol inhibits cell proliferation, invasion and migration via activating Wnt/β-catenin signaling in HBx-HepG2 cells. 相似文献
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SHANG Dan ZHENG Qi-chang SONG Zi-fang WANG Xie-dan HU Qing-gang GUO Xing-jun 《园艺学报》2007,23(10):1887-1890
AIM: To express human Arresten gene in eukaryotic cell,and to investigate its effect on the proliferation and migration in vitro of rat primary cultured thoracic aortic vascular smooth cells (VSMCs).METHODS: COS-7 cells were transfected with recombinant eukaryotic expression plasmid pSecTag2-AT or control plasmid pSecTag2 mediated by liposome.48 hours after transfection,polymerase chain reaction (RT-PCR) was used to detect the expression of Arresten mRNA in the cells,while Western blotting assay was applied to detect expressed Arresten protein in concentrated supernatants.VSMCs were then co-cultured with the concentrated supernatants;and its proliferation was detected using cell counting kit-8 (CCK-8) in vitro.Migration of VSMCs was assayed by a microchemotaxis chamber and a polycarbonate filter (Transwell's chamber) with pores of 8 μm in diameter.RESULTS: RT-PCR revealed that the genome of Arresten-transferred cells contained a 449bp specific fragment of Arresten gene.Successful protein expression in supernatants was confirmed by Western blotting.CCK-8 assay showed that the proliferation of VSMCs was inhibited significantly by Arresten protein as compared with control group (P<0.01).Transwell's chamber showed that the number of control group,pSecTag2 transfected group and pSecTag2-AT transfected group were 28.70±3.97,26.10±4.53 and 14.00±3.33 (P<0.01).CONCLUSION: Arresten protein expressed in eukaryotic cells inhibits the proliferation and migration of VSMCs effectively in vitro. 相似文献
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AIM: To investigate the role of α1 and β2 adrenoceptors(α1AR and β2AR) in the proliferation of hypoxic pulmonary artery smooth muscle cells (PASMCs).METHODS: PASMCs were isolated by an explant method from neonatal bovine pulmonary arteries. The cultured PASMCs were exposed to 6.6% O2 for 6 h, 12 h and 24 h. The method of -TdR incorporation was used to measure the proliferation of PASMCs. i was assayed with Fura-2/AM. The mRNA expression of α1AR, β2AR, c-fos and c-myc was determined by Northern blotting. The effects of activation of α1AR and β2AR, and inhibition of α1AR on the above indexes were observed by treating PASMCs with different AR agonists and antagonists under hypoxic condition.RESULTS: Significant increase in TdR incorporation in hypoxic PASMCs with α1AR activation was observed, and marked decrease in that was induced by α1AR inhibition. However, no significant change was found after β2AR activation. i , the mRNA expression of c-fos, c-myc, α1AR and β2AR in PASMCs were increased after hypoxia.CONCLUSION: Hypoxia induces the increase in i and mRNA expression of c-fos and c-myc, leading to the proliferation of PASMCs. The hypoxic proliferation of PASMCs is intervened by α1AR, but not β2AR. The remodeling of pulmonary arteriole and pulmonary hypertension may be involved in the processes of pulmonary arteriole constriction and proliferation induced by hypoxia through up-regulation of α1AR. 相似文献
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ZHANG Yi-min YE Ren-gao LI You-ji LI Xiao-yan WANG Chang-yun YU Xue-qing DONG Xiu-qing 《园艺学报》2007,23(9):1796-1800
AIM: To study the effects and mechanism of recombinant human defensin α1 on cell proliferation in cultured rat glomerular mesangial cells.METHODS: The influences of defensin α1 at various concentrations on rat 1097 mesangial cell line cultured in vitro were evaluated with MTT assay.The different concentrations of U0126,signal-regulated protein kinase (MEK) inhibitor,were added into the culture mediums of mesangial cells to do blocking test.Incubated with a final concentration of 3 mg/L defensin α1,the phosphorylation of extracellular signal regulated kinase (ERK)1/2 and type IV collagen of mesangial cells in different times were evaluated by Western blotting.RESULTS: Defensin α1 at 3-20 mg/L enhanced proliferation of rat glomerular mesangial cells.The incubation times for the maximum effect on proliferation was 12 h (P<0.01),whereas defensin α1 concentration >20 mg/L decreased cell proliferation.The cell proliferation induced by defensin α1 was inhibited by U0126.Stimulation of the cells with defensin α1 at concentration of 3 mg/L for 5 minutes induced a maximum effect on a ratio of phosphorylation of ERK1/2 to total ERK.After 12 h incubation with defensin α1,an increase in type IV collagen was observed by Western blotting and continued to increase at 24 h and 48 h (P<0.01).CONCLUSION: Defensin α1 enhances rat glomerular mesangial cell proliferation and induces type IV collagen production by MAPK signaling pathway. 相似文献
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AIM: To further investigate the role of PKARⅠβ in the growth-promoting effects of shuang long Jiegu pill (SLJGP), a Chinese medicine, on cultured osteoblasts. METHODS: pcDNA- antiPKARⅠβ, a recombinant expressing the antisense sequence of PKARⅠβ, was constructed and transformed HFOB1.19 by lipofectin. MTT was undertaken to assess the cell growth with the treatment of high dosage of SLJGP containing serum. RESULTS: Antisense gene blocked the growth-promoting effects of SLJGP containing serum on HFOB1.19. CONCLUSION: The function of SLJGP is closely related to cAMP-dependent protein kinase A. 相似文献
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AIM: To study the effect of basic fibroblast growth factor(bFGF) to the expression of integrin β1 subunit of endothelial cells. METHOD: The expression of integrin β1 subunit of endothelial cells was determined before and after bFGF treatment with Western Blot and immage analysis method.RESULTS: The mean gray value of immunostain by immage analysis method is 166.11±9.86 in the experimental group and 175.32±5.12 in the control group, suggested that bFGF may upregulate expression of integrin β1 subunits of endothelial cell. CONCLUSION: bFGF may play an important role in angiogenesis by way of inducing the overexpression of endothelial cell integrin β1 subunits. 相似文献
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ZHONG Ying-qiang HUANG Hua-rong LIU Juan LIN Ying XIA Zhong-sheng ZAN Hui 《园艺学报》2011,27(7):1290-1296
AIM: To investigate the effects of silencing of cyclooxygenase-2 (COX-2) gene expression by siRNA on the proliferation, apoptosis, cell cycle and tumorigenicity of human pancreatic cancer Capan-2 cells.METHODS: The gene transfection was performed using Lipofectamine 2000 (Lipo). The proliferation, apoptosis and cell cycle of Capan-2 cells were tested by the methods of cell counting, microscopy and FCM. The mRNA expression of COX-2 was determined by RT-PCR and real-time PCR. The protein level of COX-2 was detected by Western blotting. The tumorigenicity of Capan-2 cells transfected with siRNA-COX-2 was determined using the model of nude mice. RESULTS: Transfection efficiency of 96.47% was obtained under the conditions that the transfection volume was 2 mL, concentration of Lipo was 5 μL and that of siRNA-COX-2 was 50 nmol/L. The best sequence of siRNA-COX-2 for silencing of COX-2 gene expression was siRNA006 with the silencing rate of up to 73% 24 h after tansfection. siRNA-COX-2 slowed down the growth of Capan-2 cells 48 h after transfection (P<0.05). At time points of 48 h and 72 h after transfection, the protein expression of COX-2 was down-regulated to 67% and 61% of the normal level, the proliferation inhibition rate was 35.48% and 56.32%, and the apoptotic rate was 2.03% and 3.27%, respectively. At time points of 24 h, 48 h and 72 h after transfection, the proportion of the cells in G0/G1 phrase was 58.03%, 63.31% and 65.66%, and that of the cells in S phase was 30.27%, 24.87% and 22.2%, respectively. The mean volume and weight of tumor tissues were remarkably decreased due to the transplantation of Capan-2 cells transfected with siRNA-COX-2.CONCLUSION: siRNA-COX-2 effectively silences the expression of COX-2 gene, inhibits the growth and decreases the tumorigenicity of Capan-2 cells. 相似文献
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AIM: To investigate the effects of 1,25-dihydroxyvitamin D3 on the proliferation of passively-sensitized human airway smooth muscle cells (HASMCs), and to explore its potential role in asthmatic airway remodeling.METHODS: HASMCs were passively sensitized with 10% serum from asthmatic patients.1,25-(OH)2D3 was used as the interventor.The effect of 1,25-(OH)2D3 on the cell proliferation and its optimal concentration were determined by MTT colorimetric assay.The cell cycle analysis was performed by flow cytometry.The expression of proliferating cell nuclear antigen (PCNA) was measured by the method of immunocytochemical staining.RESULTS: 1,25-(OH)2D3 at the concentrations of 10-9-10-7 mol/L markedly inhibited the cell proliferation and the maximum effect was observed at the concentration of 10-7 mol/L.This concentration of 1,25-(OH)2D3 markedly suppressed the PCNA-positive rate and hampered the G1/S transition in HASMCs passively-sensitized by asthmatic serum.CONCLUSION: 1,25-(OH)2D3 has direct inhibitory effects on the proliferation of passively-sensitized HASMCs in vitro, which may be concerned with the beneficial role of 1,25-(OH)2D3 on the prevention and therapy of asthmatic airway remodeling. 相似文献
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DING Hao LI Ju-xiang HONG Kui SUN Guo-fang ZHANG Nan WU Yan-qing WU Qing-hua CHENG Xiao-shu 《园艺学报》2011,27(4):625-631
AIM: To investigate the effects of human xeroderma pigmentosum group D (XPD) gene on the proliferation of human vascular smooth muscle cells (VSMCs) induced by interleukin-6 (IL-6). METHODS: Recombinant plasmid pEGFP-N2/XPD and vacant plasmid pEGFP-N2 were transfected into VSMCs by liposome, and then these cells were incubated with IL-6 at 1×105 U/L for 48 h. The cells were divided into 6 groups: blank control group; pEGFP-N2 group; pEGFP-N2/XPD group; IL-6 group; IL-6 + pEGFP-N2 group; IL-6 + pEGFP-N2/XPD group. The expression of green fluorescent protein was observed under fluorescence microscope. The cell growth was detected by MTT method. The cell cycle and apoptosis rate were examined by flow cytometre. The expression levels of XPD, Bcl-2, Bax and wild type P53 (wt-P53) were detected by RT-PCR and Western blotting.RESULTS: Green fluorescence was observed in the cells transfected with pEGFP-N2/XPD or pEGFP-N2, indicating successful transfection MTT results showed that the transfection of pEGFP-N2/XPD inhibited the cell growth, and reduced the positive effects of IL-6 on VSMCs growth. Flow cytometry results showed that the transfection of pEGFP-N2/XPD increased the apoptosis rate of VSMCs and the cell numbers in G0/G1 phase, decreased the cell numbers in S phase, and reduced the effects that IL-6 decreased the apoptosis rate of VSMCs and the cell numbers in G0/G1 phase, and increased the cell numbers in S phase. The results of RT-PCR and Western blotting showed that the transfection of pEGFP-N2/XPD increased the expression of XPD, Bax and wt-P53, decreased the expression of Bcl-2, and reduced the effects that IL-6 decreased the expression of Bax and wt-P53, and increased the expression of Bcl-2. CONCLUSION: XPD gene inhibits VSMCs proliferation, promotes VSMCs apoptosis, and reduces the effects that IL-6 promotes VSMCs proliferation and inhibits VSMCs apoptosis. Therefore, XPD gene is likely to be potential molecular target for treatment of atherosclerosis. 相似文献
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AIM:To investigate the effects of xeroderma pigmentosum group D (XPD) protein on the growth of human hepatoma HepG2 cells and the expression of retinoblastoma (Rb) and mitotic arrest deficient 2 (MAD2) proteins. METHODS:The recombinant plasmid pEGFP-N2-XPD and empty plasmid pEGFP-N2 were transfected into HepG2 cells by LipofectamineTM 2000. The cells were divided into 4 groups including blank control group, liposome group, pEGFP-N2 group (N2 group) and pEGFP-N2-XPD group (XPD group). The expression of XPD, Rb and MAD2 at mRNA and protein levels was detected by RT-PCR and Western blotting. The cell growth was measured by MTT assay. The cell apoptosis and cell cycle were analyzed by flow cytometry. RESULTS:Overexpression of XPD up-regulated the expression of Rb, and down-regulated the expression of MAD2 at mRNA and protein levels. XPD inhibited the proliferation of HepG2 cells and exacerbated the apoptosis. XPD prevented the hepatoma cells from G1 stage to S stage. CONCLUSION:XPD suppresses the growth of hepatoma cells, up-regulates the expression of Rb, and down-regulates the expression of MAD2. 相似文献
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AIM and METHODS: To investigate the expression of adhesion molecule β2 integrins (CD11a、CD11b) and L-selectin (CD62L )on acute lymophocyte leukemia (ALL) cells and its clinical implications. Adhesion molecules CD11a, CD11b and CD62L of 45 ALL patients and 25 health people were measured by flow-cytometric analysis. RESULTS: ①CD11a and CD11b expression were lower on ALL cells than the normal hematopoietic cells. The rate of low expression was 100% for CD11b, 50% for CD11a, respectively. CD62L expression were higher on ALL cells than the normal hematopoietic cells. ②The CD11a was lower expressed on B-ALL than T-ALL. CD62L was higher on T-ALL than B-ALL. ③The expression of CD11a in the invasion group was much higher than that in the non-invasive group (P<0.05). ④The levels of CD11a, CD11b were returned to normal levels at remission. CONCLUSION: These results suggest that there are abnormalities in the expression of cell adhesion molecules in ALL which may help identify ALL subtypes and the treatment effect. 相似文献
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Effect of ABCE 1 gene silencing on proliferation and migration of human bladder cancer cell line T24
AIM: To investigate the effect of small interfering RNA (siRNA)-mediated ABCE1 knockdown on the survival, cell cycle and invasion of human bladder cancer cell line T24. METHODS: The siRNA against ABCE1 was constructed and transfected into the T24 cells with LipofectamineTM 2000. The expression of ABCE1 was detected by RT-PCR and Western blot. Flow cytometry was used to detect the cell cycle. The effects of ABCE1 gene silencing on proliferation, migration and invasion of T24 cells were evaluated by CCK-8 assay, wound-healing assay and Transwell invasion assay, respectively. RESULTS: Compared with control group and blank group, the mRNA and protein levels of ABCE1 were significantly decreased in experimental group after transfected with ABCE1 siRNA. The cell cycle was arrested at G0/G1 phase and the cell number in S phase was decreased in the T24 cells. Compared with control group and blank group, the proliferation of T24 cells in experimental group was inhibited significantly, and the migration and invasion abilities of T24 cells in experimental group decreased significantly. CONCLUSION: Knockdown of ABCE1 gene may decrease migration, invasion and proliferation abilities in T24 cells. 相似文献
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SU Cheng-cheng ZHANG Yi-dan MA Yong-qiang CHEN Xue-fen XIANG Guo-an ZHOU Xin PENG Shou-chun LIN Zhi-chun WEI Lu-qing JI Wen-jie 《园艺学报》2015,31(11):2065-2069
AIM: To establish a cell line of stable silencing of P2X7 receptor (P2X7R) expression through short hairpin RNA (shRNA)-mediated interference in murine RAW264.7 macrophages, and to investigate the proliferation and apoptosis in the cell line. METHODS: Stable silencing of P2X7R gene in the RAW264.7 cells was achieved by recombinant shRNA plasmid targeting murine P2X7R gene via liposome mediated transfection, followed by G418 selection. The efficacy of plasmid transfection and P2X7R silencing in G418 resistant cells was verified by immunofluorescent microscopy and real-time PCR, respectively. The proliferative activity was analyzed by CCK-8 assay and EdU cell proliferation assay. The cell cycle distribution and apoptosis were evaluated by flow cytometry. RESULTS: The expression of P2X7R at mRNA and protein levels was down-regulated by 80% in sh P2X7R group compared with negative control (NC) plasmid transfection. In addition, P2X7R-silencing cells exhibited higher proliferative activity compared with NC and wild-type RAW264.7 cells (P<0.05). Compared with NC cells, P2X7R silencing resulted in an increase in the phagocytosis of the cells (P<0.05). CONCLUSION: A cell line RAW264.7 of stable silencing of P2X7R expression was successfully established. P2X7R gene silencing stimulates the proliferation, and changes phagocytic function in murine RAW264.7 macrophages. 相似文献