AIM： To investigate the effed3 of anti-sense oligonucleotides （ASODNs） on mRNA expression of heparanase in human esophageal cancer EC9706 cells. METHODS： One non-sense oligonucleotide （N-ODN） and five ASODNs aga...AIM： To investigate the effed3 of anti-sense oligonucleotides （ASODNs） on mRNA expression of heparanase in human esophageal cancer EC9706 cells. METHODS： One non-sense oligonucleotide （N-ODN） and five ASODNs against different heparanase mRNA sites were transfected into EC9706 cells, then the expression of heparanase mRNA in EC9706 cells was studied by in situ hybridization. RESULTS： The expression of heparanase mRNA could be inhibited by ASODNs.There was no significant difference among five ASODNs （P〉0.05）, but there was a significant difference between ASODNs and N-ODN or non-transfected group （ASODNI： 2.25±0.25, ASODN2： 2.21±0.23, ASODN3： 2.23±0.23, ASODN4：2.25±0.24 vs N-ODN： 3.47±2.80 or non- transfected group： 3.51±2.93 respectively, P〈0.05）. CONCLUSION： The expression of heparanase mRNA in EC9706 cells can be inhibited by ASODNs in vivo, and heparanase ASODNs can inhibit metastasis of esophageal squamous cell carcinoma or other tumors by inhibiting the expression of heparanase.展开更多
Background:The adhesion of monocytes to the endothelium following accumulation of low-density lipoprotein (LDL) in subendothelial spaces is an important step in the development of intimal hyperplasia in arterially imp...Background:The adhesion of monocytes to the endothelium following accumulation of low-density lipoprotein (LDL) in subendothelial spaces is an important step in the development of intimal hyperplasia in arterially implanted vein grafts and atherosclerosis in both animals and humans. However, it is not well known how serum factors affect the adhesion of monocytes. Methods: We have studied the effect of fetal calf serum (FCS), which we considered a source of LDL, on the adhesion of monocytes to endothelial cells (ECs) by using human monocytic THP-1 cells and both a monolayer of cultured bovine aortic endothelial cells (EC monoculture) and a co-culture with bovine aortic smooth muscle cells (EC-SMC co-culture). Results: It was found that the addition of FCS to the medium greatly affected the adhesion of THP-1 cells, and the higher the concentration of FCS in the medium, the greater the adhesion of THP-1 cells to endothelial cells. Adhesion of THP-1 cells to an EC-SMC co-culture was approximately twofold greater than that to an EC monoculture, and after adhering to endothelial cells, many THP-1 cells trans-migrated into the layer of smooth muscle cells. Conclusion: The results suggest that the elevation of the LDL (cholesterol) level in blood provides a favorable condition for the development of intimal hyperplasia and atherosclerosis by promoting the adhesion of monocytes to the endothelium and their subsequent migration into subendothelial spaces.展开更多
Chromogranin A (CgA) is a kind of acidic protein originally isolated from the chromaffin cells of the ox adrenal gland medulla. It was found that the CgA was also widely distributed in the neuroendocrine cells besides...Chromogranin A (CgA) is a kind of acidic protein originally isolated from the chromaffin cells of the ox adrenal gland medulla. It was found that the CgA was also widely distributed in the neuroendocrine cells besides the adrenal medulla. The physiological function of the CgA is not yet elucidated, but it was presumed to be involved in organizing the granule matrix and regulating the processing of the prohormones. Besides, CgA is also the precursor of some peptide hormones such展开更多
Objective:The purpose of this study was to observe the anti-cancer activity of Tonglian decoction(TD)on esophageal cancer(EC)cells in vitro,and to elucidate the related molecular mechanisms in the cell cycle and PI3K/...Objective:The purpose of this study was to observe the anti-cancer activity of Tonglian decoction(TD)on esophageal cancer(EC)cells in vitro,and to elucidate the related molecular mechanisms in the cell cycle and PI3K/Akt signaling pathway.Methods:EC9706 cells were cultured in RPMI 1640 medium supplemented with 10%calf serum at 37C in a 5%CO2 incubator.The cells were treated with rat serum containing TD or the serum of rats administered Xiaoaiping as a positive control drug.Cell proliferation was assessed by methylthiazolyldiphenyl-tetrazolium bromide assays.Cell morphology was observed under a microscope.The cell cycle was examined by flow cytometry.Protein expression in the PI3K/Akt signaling pathway was measured by western blotting.Results:TD mainly inhibited cell proliferation.Concentrations of 50%cell inhibition by rat serum containing TD or Xiaoaiping were 73.6 and 153.8 mL/mL,respectively.TD also influenced cell morphology characterized by small shrunken cells.Cell colonies became small and the cell proliferation rate was slower.In cell cycle analysis,the percentage of cells in S phase was decreased significantly by TD and Xiaoaiping compared with the blank control group(P<.05).Western blotting showed that serum containing TD strongly downregulated EGFR,PI3K,Akt,p-Akt,and mTOR expression compared with the blank control group(P<.05).Conclusion:TD could inhibit EC9706 carcinoma cell proliferation by blocking the cell cycle progression in S phase.The possible mechanism was inhibition of multiple targets in the PI3K/Akt signaling pathway by TD.展开更多
Objective To investigate the effects of Total Saponins of Panax notoginseng(PNS) and Liguastrazine(LIT) on the proliferation of cultured cerebral microvascular endothelial cells. Methods The inverted microscope was us...Objective To investigate the effects of Total Saponins of Panax notoginseng(PNS) and Liguastrazine(LIT) on the proliferation of cultured cerebral microvascular endothelial cells. Methods The inverted microscope was used to observe endothelial cells and immunochemical methods was also used to detect FVIII-related antigens so as to observe endothelial cells. PNS or LIT in concentrations 0.5?g·L -1, 1.0?g·L -1 and 2.0?g·L -1 were used on the cultured cerebral endothelial cells of rats for 24 hours. MTT method was adopted to determine the outcome of endothelial proliferation. Results 1. Immunochemical methods was used to detect FVIII-related antigens. The brownish yellow showed positive, and the observation of the cultured endothelial cells under inverted microscope showed that the cells appeared to be in the morphological form of cobble-stones. 2. PNS in lower concentration (0.5?g·L -1) could facilitate the proliferation of the cells, while 1?g·L -1 and 2?g·L -1 of PNS could inhibit the proliferation of the cells. 0.5?g·L -1 of LIT could facilitate the proliferation of cellswhile LIT of 1?g·L -1 and 2?g·L -1 had no significant effect. Conclusion The two kind of TCM ingredients extracted in lower concentration could facilitate the proliferation of the cells. And, at the same concentration, the inhibition of PNS on the cells is stronger than that of LIT.展开更多
目的探讨用重组苦瓜蛋白MAP30(Momordica anti-HIV protein of 30 ku)诱导人食管癌细胞株EC-1.71细胞凋亡状况。方法 MAP30诱导EC-1.71细胞后,透射电镜观察细胞凋亡的形态特征;JC-1荧光探针检测细胞线粒体膜电位(ΔΨm)变化;ELISA法检...目的探讨用重组苦瓜蛋白MAP30(Momordica anti-HIV protein of 30 ku)诱导人食管癌细胞株EC-1.71细胞凋亡状况。方法 MAP30诱导EC-1.71细胞后,透射电镜观察细胞凋亡的形态特征;JC-1荧光探针检测细胞线粒体膜电位(ΔΨm)变化;ELISA法检测细胞培养上清液中细胞色素c(Cytc)和肿瘤坏死因子-α(TNF-α)的水平;免疫细胞化学法检测细胞ERK1/PERK1,AKT/PAKT和NF-κB表达;荧光微板阅读仪检测细胞caspase-12活性。结果 MAP30诱导可以使EC-1.71细胞固缩,核膜扭曲,核染色质聚集成块并靠近核膜,内质网高度膨胀、扩张;ΔΨm水平下降;Cytc和TNF-α含量增加;胞质中ERK1、AKT表达增强而胞核中PERK1、PAKT表达降低,NF-κB活性降低,caspase-12活性增强。结论 MAP30可以诱导EC-1.71细胞凋亡。展开更多
基金Supported by the Natural Science Foundation of Henan Province,No. 0311043700the Foundation for Young Mainstay Teachers in Colleges and universities of Henan Province, No.100(2003)the Building Foundation for 211 Key Fields during the 15th Five-year Plan Period of Ministry of Education, No. 2(2002)
文摘AIM： To investigate the effed3 of anti-sense oligonucleotides （ASODNs） on mRNA expression of heparanase in human esophageal cancer EC9706 cells. METHODS： One non-sense oligonucleotide （N-ODN） and five ASODNs against different heparanase mRNA sites were transfected into EC9706 cells, then the expression of heparanase mRNA in EC9706 cells was studied by in situ hybridization. RESULTS： The expression of heparanase mRNA could be inhibited by ASODNs.There was no significant difference among five ASODNs （P〉0.05）, but there was a significant difference between ASODNs and N-ODN or non-transfected group （ASODNI： 2.25±0.25, ASODN2： 2.21±0.23, ASODN3： 2.23±0.23, ASODN4：2.25±0.24 vs N-ODN： 3.47±2.80 or non- transfected group： 3.51±2.93 respectively, P〈0.05）. CONCLUSION： The expression of heparanase mRNA in EC9706 cells can be inhibited by ASODNs in vivo, and heparanase ASODNs can inhibit metastasis of esophageal squamous cell carcinoma or other tumors by inhibiting the expression of heparanase.
基金a Grant-in-Aid for Scientific Research onPriority Areas (No. 15086201) from the Ministry of Education, Culture, Sports, Science and Technology of Japanthe Health Bureauof Zhejiang Province (No. 2007B132), China
文摘Background:The adhesion of monocytes to the endothelium following accumulation of low-density lipoprotein (LDL) in subendothelial spaces is an important step in the development of intimal hyperplasia in arterially implanted vein grafts and atherosclerosis in both animals and humans. However, it is not well known how serum factors affect the adhesion of monocytes. Methods: We have studied the effect of fetal calf serum (FCS), which we considered a source of LDL, on the adhesion of monocytes to endothelial cells (ECs) by using human monocytic THP-1 cells and both a monolayer of cultured bovine aortic endothelial cells (EC monoculture) and a co-culture with bovine aortic smooth muscle cells (EC-SMC co-culture). Results: It was found that the addition of FCS to the medium greatly affected the adhesion of THP-1 cells, and the higher the concentration of FCS in the medium, the greater the adhesion of THP-1 cells to endothelial cells. Adhesion of THP-1 cells to an EC-SMC co-culture was approximately twofold greater than that to an EC monoculture, and after adhering to endothelial cells, many THP-1 cells trans-migrated into the layer of smooth muscle cells. Conclusion: The results suggest that the elevation of the LDL (cholesterol) level in blood provides a favorable condition for the development of intimal hyperplasia and atherosclerosis by promoting the adhesion of monocytes to the endothelium and their subsequent migration into subendothelial spaces.
文摘Chromogranin A (CgA) is a kind of acidic protein originally isolated from the chromaffin cells of the ox adrenal gland medulla. It was found that the CgA was also widely distributed in the neuroendocrine cells besides the adrenal medulla. The physiological function of the CgA is not yet elucidated, but it was presumed to be involved in organizing the granule matrix and regulating the processing of the prohormones. Besides, CgA is also the precursor of some peptide hormones such
基金the National Natural Science Fund of China(81101912)Natural Science Fund of Hebei Province of China(H2013209053)Scientific and Technological Project of Administration of Traditional Chinese Medicine of Hebei(2014185)。
文摘Objective:The purpose of this study was to observe the anti-cancer activity of Tonglian decoction(TD)on esophageal cancer(EC)cells in vitro,and to elucidate the related molecular mechanisms in the cell cycle and PI3K/Akt signaling pathway.Methods:EC9706 cells were cultured in RPMI 1640 medium supplemented with 10%calf serum at 37C in a 5%CO2 incubator.The cells were treated with rat serum containing TD or the serum of rats administered Xiaoaiping as a positive control drug.Cell proliferation was assessed by methylthiazolyldiphenyl-tetrazolium bromide assays.Cell morphology was observed under a microscope.The cell cycle was examined by flow cytometry.Protein expression in the PI3K/Akt signaling pathway was measured by western blotting.Results:TD mainly inhibited cell proliferation.Concentrations of 50%cell inhibition by rat serum containing TD or Xiaoaiping were 73.6 and 153.8 mL/mL,respectively.TD also influenced cell morphology characterized by small shrunken cells.Cell colonies became small and the cell proliferation rate was slower.In cell cycle analysis,the percentage of cells in S phase was decreased significantly by TD and Xiaoaiping compared with the blank control group(P<.05).Western blotting showed that serum containing TD strongly downregulated EGFR,PI3K,Akt,p-Akt,and mTOR expression compared with the blank control group(P<.05).Conclusion:TD could inhibit EC9706 carcinoma cell proliferation by blocking the cell cycle progression in S phase.The possible mechanism was inhibition of multiple targets in the PI3K/Akt signaling pathway by TD.
文摘Objective To investigate the effects of Total Saponins of Panax notoginseng(PNS) and Liguastrazine(LIT) on the proliferation of cultured cerebral microvascular endothelial cells. Methods The inverted microscope was used to observe endothelial cells and immunochemical methods was also used to detect FVIII-related antigens so as to observe endothelial cells. PNS or LIT in concentrations 0.5?g·L -1, 1.0?g·L -1 and 2.0?g·L -1 were used on the cultured cerebral endothelial cells of rats for 24 hours. MTT method was adopted to determine the outcome of endothelial proliferation. Results 1. Immunochemical methods was used to detect FVIII-related antigens. The brownish yellow showed positive, and the observation of the cultured endothelial cells under inverted microscope showed that the cells appeared to be in the morphological form of cobble-stones. 2. PNS in lower concentration (0.5?g·L -1) could facilitate the proliferation of the cells, while 1?g·L -1 and 2?g·L -1 of PNS could inhibit the proliferation of the cells. 0.5?g·L -1 of LIT could facilitate the proliferation of cellswhile LIT of 1?g·L -1 and 2?g·L -1 had no significant effect. Conclusion The two kind of TCM ingredients extracted in lower concentration could facilitate the proliferation of the cells. And, at the same concentration, the inhibition of PNS on the cells is stronger than that of LIT.
文摘目的探讨用重组苦瓜蛋白MAP30(Momordica anti-HIV protein of 30 ku)诱导人食管癌细胞株EC-1.71细胞凋亡状况。方法 MAP30诱导EC-1.71细胞后,透射电镜观察细胞凋亡的形态特征;JC-1荧光探针检测细胞线粒体膜电位(ΔΨm)变化;ELISA法检测细胞培养上清液中细胞色素c(Cytc)和肿瘤坏死因子-α(TNF-α)的水平;免疫细胞化学法检测细胞ERK1/PERK1,AKT/PAKT和NF-κB表达;荧光微板阅读仪检测细胞caspase-12活性。结果 MAP30诱导可以使EC-1.71细胞固缩,核膜扭曲,核染色质聚集成块并靠近核膜,内质网高度膨胀、扩张;ΔΨm水平下降;Cytc和TNF-α含量增加;胞质中ERK1、AKT表达增强而胞核中PERK1、PAKT表达降低,NF-κB活性降低,caspase-12活性增强。结论 MAP30可以诱导EC-1.71细胞凋亡。
