Sun G G, Sheng S H, Jing S W, Hu W N
Department of Chemoradiotherapy, Tangshan People's Hospital, No. 65, Shengli Road, Lunan District, Tangshan, 063000, Hebei Province, China.
Tumour Biol. 2014 Mar;35(3):2641-8. doi: 10.1007/s13277-013-1347-1. Epub 2013 Nov 16.
This study aimed to analyze the expression and clinical significance of filamin A (FLNA) in gastric carcinoma and the biological effect in its cell line by FLNA overexpression. Immunohistochemistry and western blot were used to analyze FLNA protein expression in 47 cases of gastric cancer and 47 cases of normal tissues to study the relationship between FLNA expression and clinical factors. FLNA lentiviral vector and empty vector were respectively transfected into gastric cancer SGC-7901 cell line. Reverse transcription-polymerase chain reaction (RT-PCR) and western blot were used to detect the mRNA level and protein of FLNA. 3-[4,5-Dimethylthiazol-2-yl]-2,5-diphenyltetrazolium bromide (MTT) assay and migration and invasion assays were also conducted to determine the influence of the upregulated expression of FLNA that might be found on SGC-7901 cell biological effect. Immunohistochemistry: The level of FLNA protein expression was found to be significantly lower in gastric cancer tissue than normal tissues (P < 0.05). Western blot: The relative amount of FLNA protein in gastric cancer tissue was found to be significantly lower than in normal tissues (P < 0.05). The level of FLNA protein expression was not correlated with gender, age, and tumor invasion (P > 0.05), but it was correlated with lymph node metastasis, clinic stage, and histological grade (P < 0.05). Loss of FLNA expression correlated significantly with poor overall survival time by Kaplan-Meier analysis (P < 0.05). The result of biological function showed that SGC-7901 cell transfected FLNA had a lower survival fraction, significant decrease in migration and invasion, and lower matrix metallopeptidase 9 (MMP-9) protein expression compared with SGC-7901 cell untransfected FLNA (P < 0.05). FLNA expression decreased in gastric cancer and correlated significantly with lymph node metastasis, clinic stage, histological grade, and poor overall survival, suggesting that FLNA may play important roles as a negative regulator to gastric cancer SGC-7901 cell by promoting degradation of MMP-9.
本研究旨在分析细丝蛋白A（FLNA）在胃癌中的表达及临床意义，以及通过FLNA过表达对其细胞系的生物学效应。采用免疫组织化学和蛋白质印迹法分析47例胃癌组织和47例正常组织中FLNA蛋白的表达情况，以研究FLNA表达与临床因素之间的关系。将FLNA慢病毒载体和空载体分别转染至胃癌SGC-7901细胞系。采用逆转录聚合酶链反应（RT-PCR）和蛋白质印迹法检测FLNA的mRNA水平和蛋白质。同时进行3-[4,5-二甲基噻唑-2-基]-2,5-二苯基四氮唑溴盐（MTT）法以及迁移和侵袭实验，以确定可能发现的FLNA表达上调对SGC-7901细胞生物学效应的影响。免疫组织化学：发现胃癌组织中FLNA蛋白表达水平明显低于正常组织（P < 0.05）。蛋白质印迹法：发现胃癌组织中FLNA蛋白的相对含量明显低于正常组织（P < 0.05）。FLNA蛋白表达水平与性别、年龄和肿瘤侵袭无关（P > 0.05），但与淋巴结转移、临床分期和组织学分级相关（P < 0.05）。通过Kaplan-Meier分析，FLNA表达缺失与总体生存时间较差显著相关（P < 0.05）。生物学功能结果显示，与未转染FLNA的SGC-7901细胞相比，转染FLNA的SGC-7901细胞的存活分数较低，迁移和侵袭能力显著降低，基质金属蛋白酶9（MMP-9）蛋白表达较低（P < 0.05）。FLNA在胃癌中表达降低，且与淋巴结转移、临床分期、组织学分级和总体生存较差显著相关，提示FLNA可能通过促进MMP-9降解作为胃癌SGC-7901细胞的负调节因子发挥重要作用。
