目的检测临床手术切除43例胃癌及相应正常组织COX-2、hMLH1和hMSH2微卫星不稳定状态MSI及三种基因启动子甲基化情况,并进一步探讨它们与胃癌发生的关系。方法采用聚合酶链反应(PCR)技术检测5个位点的MSI状态;甲基化特异性PCR(Methylatio...目的检测临床手术切除43例胃癌及相应正常组织COX-2、hMLH1和hMSH2微卫星不稳定状态MSI及三种基因启动子甲基化情况,并进一步探讨它们与胃癌发生的关系。方法采用聚合酶链反应(PCR)技术检测5个位点的MSI状态;甲基化特异性PCR(Methylation specific PCR,MSP)方法检测胃癌及正常组织COX-2、hMLH1及hMSH2三种基因启动子CpG岛甲基化状态。结果43例胃癌中MSI总检出率为48.84%(21/43),五个位点的MSI检出率无显著差别。COX-2和hMLH1基因启动子CpG岛甲基化在43例胃癌中分别有8例和13例,正常组织中未检测到。hMSH2基因启动子CpG岛在胃癌及正常组织中均未检测到甲基化。在MSI-H组hMLH1基因启动子CpG岛甲基化率显著高于MSS组(P<0.01);而在MSI-H和MSI-L组间以及MSI-L和MSS无差别。MSI-H组中COX-2基因启动子CpG岛甲基化8例,且有7例胃癌同时出现hMLH1和COX-2基因启动子CpG岛甲基化。结论在MSI胃癌(尤其是MSI-H型胃癌)的发生、发展过程中可能同时出现了hMLH1和COX-2基因启动子CpG岛的甲基化(即表型遗传修饰)。MSI胃癌hMLH1基因启动子CpG岛甲基化率高于MSS胃癌,提示检测hMLH1基因启动子CpG岛甲基化对于判断肿瘤类型有一定意义。展开更多
AIM： TO determine the possible modulating effect of the COX-2 polymorphisms, -765G→C and -1195A→G, on the risk of colorectal cancer （CRC） in a Dutch population. METHODS： This case-control study includes 326 pati...AIM： TO determine the possible modulating effect of the COX-2 polymorphisms, -765G→C and -1195A→G, on the risk of colorectal cancer （CRC） in a Dutch population. METHODS： This case-control study includes 326 patients with CRC and 369 age- and gender-matched controls. Genotypes of the COX-2 polymorphisms -7dEG→C and -1195A→G were determined by polymerase chain reaction-based restriction fragment length polymorphism. COX-2 genotypes and haplotypes were analyzed and odds ratios with 95% confidence intervals were estimated by logistic regression. RESULTS： The -765GG genotype was associated with an increased risk of developing CRC （OR, 1.45; 95% CI, 1.03-2.04）. No significant difference was observed in the genotype distribution of the -1195A→G polymorphism between patients and controls. The GG/AC haplotype was present significantly less often in patients than in controls （OR 0.44; 95% CI, 0.22-0.85）. When the AC, AG and GG haplotypes were investigated separately, the AC haplotype showed a tendency to be less frequent in patients than in controls （OR（AG/AC） 0.78; 95% CI, 0.57-1.06）. CONCLUSION： The -765GG genotype is associated with an increased risk of developing CRC and the G6/ AC haplotype seems to protect against CRC. These findings suggest a modulating role for the COX-2 polymorphisms -765G→C and -1195A→G in the development of CRC in a Dutch population.展开更多
Transient receptor potential ankyrin 1 （TRPA1） is a key player in pain and neurogenic inflammation, and is localized in nociceptive primary sensory dorsal root ganglion （DRG） neurons. TRPA1 plays a major role in t...Transient receptor potential ankyrin 1 （TRPA1） is a key player in pain and neurogenic inflammation, and is localized in nociceptive primary sensory dorsal root ganglion （DRG） neurons. TRPA1 plays a major role in the transmission of nociceptive sensory signals. The generation of neurogenic inflammation appears to involve TRPAl-evoked release of calcitonin gene-related peptide （CGRP）. However, it remains unknown whether TRPA1 or CGRP expression is affected by TRPA 1 activation. Thus, in this study, we examined TRPA 1 and CGRP expression in DRG neurons in vitro after treatment with the TRPA1 activator tbrmaldehyde or the TRPA1 blocker menthol. In addition, we examined the role of extracellular signal-regulated protein kinase 1/2 （ERK1/2） in this process. DRG neurons in culture were exposed to formaldehyde, menthol, the ERK1/2 inhibitor PD98059 ＋ formaldehyde, or PD98059 ＋ menthol. After treatment, real-time polymerase chain reaction, western blot assay and double immunofluorescence labeling were performed to evaluate TRPA1 and CGRP expression in DRG neurons. Formaldehyde elevated mRNA and protein levels of TRPA 1 and CGRP, as well as the proportion of TRPA1- and CGRP-positive neurons. In contrast, menthol reduced TRPA1 and CGRP expression. Furthermore, the effects of lbrmaldehyde, but not menthol, on CGRP expression were blocked by pretreatment with PD98059. PD98059 pretreatment did not affect TRPA1 expression in the presence of formaldehyde or menthol.展开更多
AIM:To investigate the mechanisms of the biological roles of Dickkopf-3(Dkk-3) in cell invasion,survival and apoptosis in colon cancer cells.METHODS:Three human colon cancer cell lines,i.e.,HT-29,LoVo and SW480,were u...AIM:To investigate the mechanisms of the biological roles of Dickkopf-3(Dkk-3) in cell invasion,survival and apoptosis in colon cancer cells.METHODS:Three human colon cancer cell lines,i.e.,HT-29,LoVo and SW480,were used.Overexpression of Dkk-3 induced by pEGFP-N1-Dkk-3-GFP plasmid in LoVo cells was performed using Lipofectamine 2000 reagent.Reverse transcription polymerase chain reaction and Western blotting were performed to determine the mRNA and protein expression levels of Dkk-3,respectively.Cell proliferation assay,cell cycle analysis,hoechst 33258 assay and Matrigel invasion assay were performed on Dkk-3 overexpressing transfectants.RESULTS:The mRNA and protein expressions of Dkk-3 in HT-29(mRNA:0.06 ± 0.02,protein:0.06 ± 0.01) and LoVo(mRNA:0.07 ± 0.02,protein:0.07 ± 0.02) cells were significantly lower than that in SW480 cells(mRNA:0.92 ± 0.04,protein:0.69 ± 0.13;all P < 0.05),and the greatest levels of invasiveness wasin LoVo cells.Dkk-3 overexpression inhibited the proliferation and invasion of LoVo cells and induced cell cycle arrest at G0/G1 phase and subsequent apoptosis,as indicated by increased chromatin condensation and fragments,upregulated Bax and cytochrome c protein,downregulated survivin and Bcl-2 protein,and the activation of caspase-3 and caspase-9.Furthermore,Dkk-3 overexpression reduced the accumulation of cytosolic fraction of β-catenin.CONCLUSION:Dkk-3 overexpression induced apoptosis in human colon cancer possibly through the mitochondrial pathway.Dkk-3 may be involved in the Wnt/β-catenin signaling pathways in colon cancer.展开更多
AIM: To investigate the effects of resistin-like molecule β (RELMβ) over-expression on the invasion, metastasis and angiogenesis of gastric cancer cells. METHODS: Human RELMβ encoding expression vec tor was constru...AIM: To investigate the effects of resistin-like molecule β (RELMβ) over-expression on the invasion, metastasis and angiogenesis of gastric cancer cells. METHODS: Human RELMβ encoding expression vec tor was constructed and transfected into the RELMβ lowly-expressed gastric cancer cell lines SGC7901 and MKN-45. Gene expression was measured by Western blotting, reverse transcription polymerase chain reaction (PCR) and real-time quantitative PCR. Cell proliferation was measured by 2-(4,5-dimethyltriazol-2-yl)-2,5-diphenyl tetrazolium bromide colorimetry, colony formation and 5-ethynyl-20-deoxyuridine incorporation assays. The in vitro migration, invasion and metastasis of cancer cells were measured by cell adhesion assay, scratch assay and matrigel invasion assay. The angiogenic capabilities of cancer cells were measured by tube formation of endothelial cells. RESULTS: Transfection of RELMβ vector into SGC-7901 and MKN-45 cells resulted in over-expression of RELMβ, which did not infl uence the cellular proliferation. However, over-expression of RELMβ suppressed the in vitro adhesion, invasion and metastasis of cancer cells, accompanied by decreased expression of matrix metalloproteinase-2 (MMP-2) and MMP-9. Moreover, transfection of RELMβ attenuated the expression of vascular endothelial growth factor and in vitro angiogenic capabilities of cancer cells. CONCLUSION: Over-expression of RELMβ abolishes the invasion, metastasis and angiogenesis of gastric cancer cells in vitro, suggesting its potentials as a novel therapeutic target for gastric cancer.展开更多
Objective: To make a clinical analysis on the basis of 36 cases of atypical genital herpes (GH) patients. Methods: Thirty-six cases of atypical GH were diagnosedclinically, and their case histories, symptoms and signs...Objective: To make a clinical analysis on the basis of 36 cases of atypical genital herpes (GH) patients. Methods: Thirty-six cases of atypical GH were diagnosedclinically, and their case histories, symptoms and signs wererecorded in detail and followed up. Polymerase chain reaction(PCR) was adopted for testing HSV2-DNA with cotton-tippedswabs. Enzyme-linked immuno sorbent assay (ELISA) forserum anti-HSV2-IgM was done to establish a definitivediagnosis. Other diagnoses were excluded at the same time bytesting for related pathogens including fungi, Chlamydia,Mycoplasma, Treponema pallidum, gonococci, Trichomonas,etc. Results: The main clinical manifestations of atypical GHwere: (1) small genital ulcers; (2) inflammation of urethralmeatus; (3) nonspecific genital erythema; (4) papuloid noduleson the glands; (5) nonspecific vaginitis. Twenty-three cases(64%) tested by PCR were HSV2-DNA sera-positive, and 36cases (100%) anti-HSV2-1gM sera-positive by ELISA. Conclusion: atypical HSV is difficult to be dinosed. But the combination of PCR and ELIAS will be helpful to thediagnosis of atypical HSV.展开更多
文摘目的检测临床手术切除43例胃癌及相应正常组织COX-2、hMLH1和hMSH2微卫星不稳定状态MSI及三种基因启动子甲基化情况,并进一步探讨它们与胃癌发生的关系。方法采用聚合酶链反应(PCR)技术检测5个位点的MSI状态;甲基化特异性PCR(Methylation specific PCR,MSP)方法检测胃癌及正常组织COX-2、hMLH1及hMSH2三种基因启动子CpG岛甲基化状态。结果43例胃癌中MSI总检出率为48.84%(21/43),五个位点的MSI检出率无显著差别。COX-2和hMLH1基因启动子CpG岛甲基化在43例胃癌中分别有8例和13例,正常组织中未检测到。hMSH2基因启动子CpG岛在胃癌及正常组织中均未检测到甲基化。在MSI-H组hMLH1基因启动子CpG岛甲基化率显著高于MSS组(P<0.01);而在MSI-H和MSI-L组间以及MSI-L和MSS无差别。MSI-H组中COX-2基因启动子CpG岛甲基化8例,且有7例胃癌同时出现hMLH1和COX-2基因启动子CpG岛甲基化。结论在MSI胃癌(尤其是MSI-H型胃癌)的发生、发展过程中可能同时出现了hMLH1和COX-2基因启动子CpG岛的甲基化(即表型遗传修饰)。MSI胃癌hMLH1基因启动子CpG岛甲基化率高于MSS胃癌,提示检测hMLH1基因启动子CpG岛甲基化对于判断肿瘤类型有一定意义。
文摘AIM： TO determine the possible modulating effect of the COX-2 polymorphisms, -765G→C and -1195A→G, on the risk of colorectal cancer （CRC） in a Dutch population. METHODS： This case-control study includes 326 patients with CRC and 369 age- and gender-matched controls. Genotypes of the COX-2 polymorphisms -7dEG→C and -1195A→G were determined by polymerase chain reaction-based restriction fragment length polymorphism. COX-2 genotypes and haplotypes were analyzed and odds ratios with 95% confidence intervals were estimated by logistic regression. RESULTS： The -765GG genotype was associated with an increased risk of developing CRC （OR, 1.45; 95% CI, 1.03-2.04）. No significant difference was observed in the genotype distribution of the -1195A→G polymorphism between patients and controls. The GG/AC haplotype was present significantly less often in patients than in controls （OR 0.44; 95% CI, 0.22-0.85）. When the AC, AG and GG haplotypes were investigated separately, the AC haplotype showed a tendency to be less frequent in patients than in controls （OR（AG/AC） 0.78; 95% CI, 0.57-1.06）. CONCLUSION： The -765GG genotype is associated with an increased risk of developing CRC and the G6/ AC haplotype seems to protect against CRC. These findings suggest a modulating role for the COX-2 polymorphisms -765G→C and -1195A→G in the development of CRC in a Dutch population.
基金supported by the National Natural Science Foundation of China,No.81501935(to HL)the Natural Science Foundation of Shandong Province of China,No.ZR2014HQ065(to HL)
文摘Transient receptor potential ankyrin 1 （TRPA1） is a key player in pain and neurogenic inflammation, and is localized in nociceptive primary sensory dorsal root ganglion （DRG） neurons. TRPA1 plays a major role in the transmission of nociceptive sensory signals. The generation of neurogenic inflammation appears to involve TRPAl-evoked release of calcitonin gene-related peptide （CGRP）. However, it remains unknown whether TRPA1 or CGRP expression is affected by TRPA 1 activation. Thus, in this study, we examined TRPA 1 and CGRP expression in DRG neurons in vitro after treatment with the TRPA1 activator tbrmaldehyde or the TRPA1 blocker menthol. In addition, we examined the role of extracellular signal-regulated protein kinase 1/2 （ERK1/2） in this process. DRG neurons in culture were exposed to formaldehyde, menthol, the ERK1/2 inhibitor PD98059 ＋ formaldehyde, or PD98059 ＋ menthol. After treatment, real-time polymerase chain reaction, western blot assay and double immunofluorescence labeling were performed to evaluate TRPA1 and CGRP expression in DRG neurons. Formaldehyde elevated mRNA and protein levels of TRPA 1 and CGRP, as well as the proportion of TRPA1- and CGRP-positive neurons. In contrast, menthol reduced TRPA1 and CGRP expression. Furthermore, the effects of lbrmaldehyde, but not menthol, on CGRP expression were blocked by pretreatment with PD98059. PD98059 pretreatment did not affect TRPA1 expression in the presence of formaldehyde or menthol.
基金Supported by The Fundamental Research Funds for the Central Universities of China,No.20103020101000197
文摘AIM:To investigate the mechanisms of the biological roles of Dickkopf-3(Dkk-3) in cell invasion,survival and apoptosis in colon cancer cells.METHODS:Three human colon cancer cell lines,i.e.,HT-29,LoVo and SW480,were used.Overexpression of Dkk-3 induced by pEGFP-N1-Dkk-3-GFP plasmid in LoVo cells was performed using Lipofectamine 2000 reagent.Reverse transcription polymerase chain reaction and Western blotting were performed to determine the mRNA and protein expression levels of Dkk-3,respectively.Cell proliferation assay,cell cycle analysis,hoechst 33258 assay and Matrigel invasion assay were performed on Dkk-3 overexpressing transfectants.RESULTS:The mRNA and protein expressions of Dkk-3 in HT-29(mRNA:0.06 ± 0.02,protein:0.06 ± 0.01) and LoVo(mRNA:0.07 ± 0.02,protein:0.07 ± 0.02) cells were significantly lower than that in SW480 cells(mRNA:0.92 ± 0.04,protein:0.69 ± 0.13;all P < 0.05),and the greatest levels of invasiveness wasin LoVo cells.Dkk-3 overexpression inhibited the proliferation and invasion of LoVo cells and induced cell cycle arrest at G0/G1 phase and subsequent apoptosis,as indicated by increased chromatin condensation and fragments,upregulated Bax and cytochrome c protein,downregulated survivin and Bcl-2 protein,and the activation of caspase-3 and caspase-9.Furthermore,Dkk-3 overexpression reduced the accumulation of cytosolic fraction of β-catenin.CONCLUSION:Dkk-3 overexpression induced apoptosis in human colon cancer possibly through the mitochondrial pathway.Dkk-3 may be involved in the Wnt/β-catenin signaling pathways in colon cancer.
基金Supported by The National Natural Science Foundation of China, No. 30200284, No. 30600278, No. 30772359, No. 81071997 and No. 81072073Program for New Century Excellent Talents from Universities, No. NCET-06-0641+1 种基金Scientific Research Fund for the Returned Overseas Chinese Scholars, No. 2008-889Fundamental Research Funds for the Central Universities, No. 2010JC025
文摘AIM: To investigate the effects of resistin-like molecule β (RELMβ) over-expression on the invasion, metastasis and angiogenesis of gastric cancer cells. METHODS: Human RELMβ encoding expression vec tor was constructed and transfected into the RELMβ lowly-expressed gastric cancer cell lines SGC7901 and MKN-45. Gene expression was measured by Western blotting, reverse transcription polymerase chain reaction (PCR) and real-time quantitative PCR. Cell proliferation was measured by 2-(4,5-dimethyltriazol-2-yl)-2,5-diphenyl tetrazolium bromide colorimetry, colony formation and 5-ethynyl-20-deoxyuridine incorporation assays. The in vitro migration, invasion and metastasis of cancer cells were measured by cell adhesion assay, scratch assay and matrigel invasion assay. The angiogenic capabilities of cancer cells were measured by tube formation of endothelial cells. RESULTS: Transfection of RELMβ vector into SGC-7901 and MKN-45 cells resulted in over-expression of RELMβ, which did not infl uence the cellular proliferation. However, over-expression of RELMβ suppressed the in vitro adhesion, invasion and metastasis of cancer cells, accompanied by decreased expression of matrix metalloproteinase-2 (MMP-2) and MMP-9. Moreover, transfection of RELMβ attenuated the expression of vascular endothelial growth factor and in vitro angiogenic capabilities of cancer cells. CONCLUSION: Over-expression of RELMβ abolishes the invasion, metastasis and angiogenesis of gastric cancer cells in vitro, suggesting its potentials as a novel therapeutic target for gastric cancer.
文摘Objective: To make a clinical analysis on the basis of 36 cases of atypical genital herpes (GH) patients. Methods: Thirty-six cases of atypical GH were diagnosedclinically, and their case histories, symptoms and signs wererecorded in detail and followed up. Polymerase chain reaction(PCR) was adopted for testing HSV2-DNA with cotton-tippedswabs. Enzyme-linked immuno sorbent assay (ELISA) forserum anti-HSV2-IgM was done to establish a definitivediagnosis. Other diagnoses were excluded at the same time bytesting for related pathogens including fungi, Chlamydia,Mycoplasma, Treponema pallidum, gonococci, Trichomonas,etc. Results: The main clinical manifestations of atypical GHwere: (1) small genital ulcers; (2) inflammation of urethralmeatus; (3) nonspecific genital erythema; (4) papuloid noduleson the glands; (5) nonspecific vaginitis. Twenty-three cases(64%) tested by PCR were HSV2-DNA sera-positive, and 36cases (100%) anti-HSV2-1gM sera-positive by ELISA. Conclusion: atypical HSV is difficult to be dinosed. But the combination of PCR and ELIAS will be helpful to thediagnosis of atypical HSV.
