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通过慢病毒介导的RNA干扰靶向CRMP-4可抑制SW480细胞增殖和结直肠癌生长。

Targeting CRMP-4 by lentivirus-mediated RNA interference inhibits SW480 cell proliferation and colorectal cancer growth.

作者信息

Chen Si-Le, Cai Shi-Rong, Zhang Xin-Hua, Li Wen-Feng, Zhai Er-Tao, Peng Jian-Jun, Wu Hui, Chen Chuang-Qi, Ma Jin-Ping, Wang Zhao, He Yu-Long

机构信息

Department of Gastrointestinal and Pancreatic Surgery, The First Affiliated Hospital of Sun Yat-sen University, Guangzhou, Guangdong 510080, P.R. China.

出版信息

Exp Ther Med. 2016 Oct;12(4):2003-2008. doi: 10.3892/etm.2016.3588. Epub 2016 Aug 10.

Abstract

The aim of the present study was to investigate the expression level of collapsin response mediator protein 4 (CRMP-4) in human colorectal cancer (CRC) tissue and to evauluate its impact on SW480 cell proliferation, in addition to tumor growth in a mouse xenograft model. Clinical CRC tissue samples were collected to detect the CRMP-4 protein expression levels using western blot and immunohistochemistry analyses. A specific small interfering RNA sequence targeting the CRMP-4 gene () was constructed and transfected into an SW480 cell line using a lentivirus vector to obtain a stable cell line with low expression of CRMP-4. The effectiveness of the interference was evaluated using western blot and reverse transcription-quantitative polymerase chain reaction, and the cell proliferation was determined using MTT and BrdU colorimetric methods. Tumor growth was assessed by subcutaneously inoculating the constructed cells into BALB/c nude mice. The protein expression levels of CRMP-4 were markedly increased in colon tumor tissue of the human samples. The proliferation of SW480 cells and the tumor growth rate in nude mice of the si-CPMR-4 group were evidently depressed compared with the si-scramble group. Thus, the present results suggest that CRMP-4 may be involved in the pathogenesis of CRC.

摘要

本研究的目的是调查塌陷反应介导蛋白4(CRMP-4)在人结直肠癌(CRC)组织中的表达水平,并评估其对SW480细胞增殖的影响以及对小鼠异种移植模型中肿瘤生长的影响。收集临床CRC组织样本,采用蛋白质印迹法和免疫组织化学分析法检测CRMP-4蛋白表达水平。构建靶向CRMP-4基因的特异性小干扰RNA序列,并使用慢病毒载体将其转染到SW480细胞系中,以获得CRMP-4低表达的稳定细胞系。采用蛋白质印迹法和逆转录-定量聚合酶链反应评估干扰效果,采用MTT和BrdU比色法测定细胞增殖情况。通过将构建的细胞皮下接种到BALB/c裸鼠中来评估肿瘤生长情况。人样本的结肠肿瘤组织中CRMP-4的蛋白表达水平显著升高。与干扰序列对照组相比,si-CPMR-4组SW480细胞的增殖和裸鼠的肿瘤生长速率明显受到抑制。因此,目前的结果表明CRMP-4可能参与了CRC的发病机制。

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