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过表达CSRP1基因可促进人肺腺癌H1299细胞的增殖、迁移和侵袭 被引量:3
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作者 张笑 余涛 +3 位作者 林河春 耿沁 潘洪玉 姚明 《肿瘤》 CAS CSCD 北大核心 2018年第4期291-299,共9页
目的:探讨半胱氨酸和甘氨酸丰富蛋白1(cysteine and glycine rich protein 1,CSRP 1)基因过表达对人肺腺癌H1299细胞增殖、周期、迁移和侵袭的影响,及其可能的作用机制。方法:构建携带有CSRP 1基因的重组慢病毒质粒pCDH-CSRP1;将重组慢... 目的:探讨半胱氨酸和甘氨酸丰富蛋白1(cysteine and glycine rich protein 1,CSRP 1)基因过表达对人肺腺癌H1299细胞增殖、周期、迁移和侵袭的影响,及其可能的作用机制。方法:构建携带有CSRP 1基因的重组慢病毒质粒pCDH-CSRP1;将重组慢病毒Ad-CDH-CSRP1和pCDH-GFP(空载体对照组)分别感染人肺腺癌H1299细胞。实时荧光定量PCR及蛋白质印迹法检测CSRP1 mRNA及蛋白在H1299细胞中的表达水平。CCK-8法和克隆形成实验检测CSRP 1基因过表达对H1299细胞增殖能力的影响;FCM法检测CSRP 1基因过表达对H1299细胞周期的影响。Transwell小室迁移实验和划痕愈合实验检测CSRP 1基因过表达对H1299细胞纵向和横向迁移能力的影响,Transwell小室侵袭实验检测对细胞侵袭能力的影响。蛋白质印迹法检测CSRP 1基因过表达对H1299细胞中黏着斑激酶(focal adhesion kinase,FAK)和磷酸化FAK(phospho-FAK,p-FAK)蛋白表达水平的影响。结果:重组慢病毒质粒pCDH-CSRP1构建成功。转入重组慢病毒质粒pCDH-CSRP1的H1299细胞中CSRP1蛋白的表达水平较pCDH-GFP组明显提高(P<0.01)。CSRP 1基因过表达可促进H1299细胞增殖(P<0.05),G1期细胞所占百分比明显下降(P<0.01),S期细胞所占百分比明显上升(P<0.01);CSRP 1基因过表达可明显增强H1299细胞的迁移和侵袭能力(P值均<0.01)。CSRP 1基因过表达的H1299细胞中p-FAK蛋白的表达水平明显升高(P<0.01),而总FAK蛋白的表达水平无明显变化(P>0.05)。结论:CSRP 1基因过表达可增强人肺腺癌H1299细胞的增殖、迁移和侵袭能力,其机制可能与FAK信号转导通路的激活有关。 展开更多
关键词 肺肿瘤 细胞增殖 细胞运动 细胞周期 csrp1 基因
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Screening of tumor suppressor genes on 1q31.1-32.1 in Chinese patients with sporadic colorectal cancer 被引量:8
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作者 ZHOU Chong-zhi QIU Guo-qiang +8 位作者 WANG Xiao-liang FAN Jun-wei TANG Hua-mei SUN Yu-hao WANG Quan HUANG Fei YAN Dong-wang LI Da-wei PENG Zhi-hai 《Chinese Medical Journal》 SCIE CAS CSCD 2008年第24期2479-2486,共8页
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
关键词 tumor suppressor gene sporadic colorectal cancer loss of heterozygosity csrp1
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