目的研究CSRP1(cysteine and glycine rich protein 1)蛋白在大肠癌组织中的表达情况,并探究其临床意义。方法 CSRP1蛋白的差异表达情况应用色谱及质谱联用技术分析筛选,初筛实验的准确性验证选用免疫印迹试验。结果分析质谱鉴定相关结...目的研究CSRP1(cysteine and glycine rich protein 1)蛋白在大肠癌组织中的表达情况,并探究其临床意义。方法 CSRP1蛋白的差异表达情况应用色谱及质谱联用技术分析筛选,初筛实验的准确性验证选用免疫印迹试验。结果分析质谱鉴定相关结果显示,CSRP1蛋白在大肠癌组织中的表达明显低于其在正常组织中的表达,免疫印迹试验结果与质谱分析结果吻合,确保了其准确性。结论 CSRP1蛋白在大肠癌组织中下调表达,其表达情况与肿瘤的发展、调节有密切的关系。展开更多
目的检测前列腺癌组织和细胞中富含半胱氨酸和甘氨酸蛋白1(cysteine and glycine-rich protein 1,CSRP1)的表达水平,并探讨其表达对前列腺癌细胞侵袭转移的影响及其机制。方法采用免疫组织化学法和实时荧光定量PCR分别检测前列腺癌组织...目的检测前列腺癌组织和细胞中富含半胱氨酸和甘氨酸蛋白1(cysteine and glycine-rich protein 1,CSRP1)的表达水平,并探讨其表达对前列腺癌细胞侵袭转移的影响及其机制。方法采用免疫组织化学法和实时荧光定量PCR分别检测前列腺癌组织和细胞系(PC-3和LNCaP)中CSRP1的表达;将CSRP1基因小干扰RNA转染PC-3细胞,下调细胞中CSRP1的表达,采用Transwell小室试验和划痕试验分别检测PC-3细胞的侵袭和迁移能力,Western blot法检测内皮间质转化(epithelial-mesenchymal transition,EMT)标志物E-cadherin和Vimentin的表达水平。结果CSRP1在前列腺癌组织中呈高表达;与正常前列腺上皮细胞RWPE-1相比,CSRP1在2种不同的前列腺癌细胞系中表达水平显著升高(P<0.05)。干扰CSRP1表达后,PC-3细胞的侵袭和迁移能力显著减弱(P<0.05),且E-cadherin的表达水平显著升高(P<0.05),Vimentin的表达受到抑制(P<0.05)。结论CSRP1在前列腺癌组织和细胞系中表达上调。沉默CSRP1表达后可下调Vimentin,逆转EMT的发生,从而抑制前列腺癌细胞的侵袭转移。CSRP1可能是前列腺癌的治疗靶点和潜在的诊断标志物。展开更多
Background As a model for both multistep and multipathway carcinogenesis, colorectal neoplastic progression provides paradigms for researching both oncogenes and tumor suppressor genes （TSGs）. However, the mechanism...Background As a model for both multistep and multipathway carcinogenesis, colorectal neoplastic progression provides paradigms for researching both oncogenes and tumor suppressor genes （TSGs）. However, the mechanism of colorectal cancer （CRC） is not completely understood, and many genes may be involved in the colorectal carcinogenesis. The purpose of this study was to screen for the potential TSGs on chromosome 1q31.1-32.1 in Chinese patients with sporadic colorectal cancer, to explore whether colorectal cancer in the Chinese population has unique genetic alterations and determine whether other putative TSGs exist and contribute to colon carcinogenesis. Methods Six polymorphic microsatellite markers, at a density of approximately one marker in every 1.6 cM, were chosen for refined loss of heterozygosity （LOH） mapping of 1q31.1-32.1. Eighty-three colorectal cancer patients＇ tumor and normal DNA were analyzed via polymerase chain reaction （PCR） for these microsatellite markers. PCR products were eletrophoresed on an ABI 377 DNA sequencer. Genescan 3.1 and Genotype 2.1 software were used for LOH scanning and analysis. On the basis of refined LOH mapping results, we undertook a microarray-based expression screening to identify tumor association genes in 19 of the CRC cases. Results The average LOH frequency of 1q31.1-32.1 was 24.41%, with the highest frequency of 36.73% （18/49） at D1S2622, and the lowest of 16.42% （11/67） at D1S412. A minimal region of frequent deletion was located within a 2 cM genomic segment at D1S413-D1S2622. There was no significant association between LOH of any marker in the studied regions and the clinicopathological data （patient sex, age, tumor size, growth pattern, or Dukes stage）. On the basis of refined mapping results, we chose 25 genes located in the D1S413-D1S2622 （1q31.3-32.1） region and presented a microarray-based high throughput screening approach in 19 sporadic CRC cases to identify candidate CRC related tumor suppressor genes. This study found 4 significantly down-expressed genes, including CSRP1, LMOD1, PPP1R12B and CFHL3. There was no significant association between expression levels of CFHL3, CSRP1, LMOD1, PPP1R12B and the clinicopathological data. By database searching, CSRP1 was hypothesized to be a colorectal cancer related tumor suppressor gene. Conclusions Through detailed deletion mapping, we found that the 1q31.3-32.1 region might harbor one or more colorectal cancer related tumor suppressor gene（s）. And by microarray-based high-throughput screening of candidate genes located in this region and by subsequent database searching, we present the first evidence that CSRP1 might be involved in the progression of CRC.展开更多
文摘目的研究CSRP1(cysteine and glycine rich protein 1)蛋白在大肠癌组织中的表达情况,并探究其临床意义。方法 CSRP1蛋白的差异表达情况应用色谱及质谱联用技术分析筛选,初筛实验的准确性验证选用免疫印迹试验。结果分析质谱鉴定相关结果显示,CSRP1蛋白在大肠癌组织中的表达明显低于其在正常组织中的表达,免疫印迹试验结果与质谱分析结果吻合,确保了其准确性。结论 CSRP1蛋白在大肠癌组织中下调表达,其表达情况与肿瘤的发展、调节有密切的关系。
文摘目的检测前列腺癌组织和细胞中富含半胱氨酸和甘氨酸蛋白1(cysteine and glycine-rich protein 1,CSRP1)的表达水平,并探讨其表达对前列腺癌细胞侵袭转移的影响及其机制。方法采用免疫组织化学法和实时荧光定量PCR分别检测前列腺癌组织和细胞系(PC-3和LNCaP)中CSRP1的表达;将CSRP1基因小干扰RNA转染PC-3细胞,下调细胞中CSRP1的表达,采用Transwell小室试验和划痕试验分别检测PC-3细胞的侵袭和迁移能力,Western blot法检测内皮间质转化(epithelial-mesenchymal transition,EMT)标志物E-cadherin和Vimentin的表达水平。结果CSRP1在前列腺癌组织中呈高表达;与正常前列腺上皮细胞RWPE-1相比,CSRP1在2种不同的前列腺癌细胞系中表达水平显著升高(P<0.05)。干扰CSRP1表达后,PC-3细胞的侵袭和迁移能力显著减弱(P<0.05),且E-cadherin的表达水平显著升高(P<0.05),Vimentin的表达受到抑制(P<0.05)。结论CSRP1在前列腺癌组织和细胞系中表达上调。沉默CSRP1表达后可下调Vimentin,逆转EMT的发生,从而抑制前列腺癌细胞的侵袭转移。CSRP1可能是前列腺癌的治疗靶点和潜在的诊断标志物。
基金This study was supported by grants from the National Natural Science Foundation of China （No. 30700813, No. 30470977, No, 30080016）, Fundamental Key Science Foundation of Science and Technology Commission of Shanghai Municipality （No. 05JC14029）, Public Scientific Sesearch Platform of Hospital of Grade A at the Tertiary Level of Shanghai （No. SHDC12007704） and Youth Science Foundation of Shanghai Health Bureau （No. 034Y03）.Acknowledgments： We thank CapitalBio Corporation for performing microarray scanning and Jacqueline Ramirez, Keith Mitchilson for editorial comments.
文摘Background As a model for both multistep and multipathway carcinogenesis, colorectal neoplastic progression provides paradigms for researching both oncogenes and tumor suppressor genes （TSGs）. However, the mechanism of colorectal cancer （CRC） is not completely understood, and many genes may be involved in the colorectal carcinogenesis. The purpose of this study was to screen for the potential TSGs on chromosome 1q31.1-32.1 in Chinese patients with sporadic colorectal cancer, to explore whether colorectal cancer in the Chinese population has unique genetic alterations and determine whether other putative TSGs exist and contribute to colon carcinogenesis. Methods Six polymorphic microsatellite markers, at a density of approximately one marker in every 1.6 cM, were chosen for refined loss of heterozygosity （LOH） mapping of 1q31.1-32.1. Eighty-three colorectal cancer patients＇ tumor and normal DNA were analyzed via polymerase chain reaction （PCR） for these microsatellite markers. PCR products were eletrophoresed on an ABI 377 DNA sequencer. Genescan 3.1 and Genotype 2.1 software were used for LOH scanning and analysis. On the basis of refined LOH mapping results, we undertook a microarray-based expression screening to identify tumor association genes in 19 of the CRC cases. Results The average LOH frequency of 1q31.1-32.1 was 24.41%, with the highest frequency of 36.73% （18/49） at D1S2622, and the lowest of 16.42% （11/67） at D1S412. A minimal region of frequent deletion was located within a 2 cM genomic segment at D1S413-D1S2622. There was no significant association between LOH of any marker in the studied regions and the clinicopathological data （patient sex, age, tumor size, growth pattern, or Dukes stage）. On the basis of refined mapping results, we chose 25 genes located in the D1S413-D1S2622 （1q31.3-32.1） region and presented a microarray-based high throughput screening approach in 19 sporadic CRC cases to identify candidate CRC related tumor suppressor genes. This study found 4 significantly down-expressed genes, including CSRP1, LMOD1, PPP1R12B and CFHL3. There was no significant association between expression levels of CFHL3, CSRP1, LMOD1, PPP1R12B and the clinicopathological data. By database searching, CSRP1 was hypothesized to be a colorectal cancer related tumor suppressor gene. Conclusions Through detailed deletion mapping, we found that the 1q31.3-32.1 region might harbor one or more colorectal cancer related tumor suppressor gene（s）. And by microarray-based high-throughput screening of candidate genes located in this region and by subsequent database searching, we present the first evidence that CSRP1 might be involved in the progression of CRC.
