Sun G G, Wang Y D, Lu Y F, Hu W N
Department of Chemoradiotherapy, Tangshan People's Hospital, NO.65, Shengli road, Lunan district, Tangshan, 063000, Hebei Province, China.
Tumour Biol. 2014 Apr;35(4):3347-54. doi: 10.1007/s13277-013-1441-4. Epub 2014 Jan 9.
This study aimed to analyze the expression, clinical significance of epithelial membrane protein 1 (EMP1) in breast carcinoma and the biological effect in its cell line by EMP1 overexpression. Immunohistochemistry and western blot were used to analyze EMP1 protein expression in 67 cases of breast cancer and 35 cases of normal tissues to study the relationship between EMP1 expression and clinical factors. Quantitative real-time RT-PCR and western blot were used to detect the mRNA level and protein of EMP1. MTT assay, migration and invasion assays were also conducted as to the influence of the upregulated expression of EMP1 that might be found on MCF-7 cell biological effect. The relative amount of EMP1 protein in breast cancer tissue was found to be significantly lower than in normal tissues (P < 0.05). The level of EMP1 protein expression was correlated with T stages, lymph node metastasis, clinic stage, and histological grade (P < 0.05). Meanwhile, loss of EMP1 expression correlated significantly with poor overall survival time by Kaplan-Meier analysis (P < 0.05). The result shown that MCF-7 cell transfected EMP1 had a lower survival fraction, higher cell apoptosis, significant decrease in migration and invasion, higher caspase-9, and lower VEGFC protein expression compared with MCF-7 cell untransfected EMP1 (P < 0.05). EMP1 expression decreased in breast cancer and correlated significantly with lymph node metastasis, clinic stage, histological grade, and poor overall survival, T stages, suggesting that EMP1 may play important roles as a negative regulator to breast cancer MCF-7 cell by regulating the expression of caspase 9 and VEGFC protein.
本研究旨在分析上皮膜蛋白1(EMP1)在乳腺癌中的表达、临床意义以及通过EMP1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67例乳腺癌组织和35例正常组织中EMP1蛋白的表达,以研究EMP1表达与临床因素之间的关系。运用定量实时逆转录-聚合酶链反应(qRT-PCR)和蛋白质印迹法检测EMP1的mRNA水平和蛋白。同时进行MTT法、迁移和侵袭实验,以探讨EMP1表达上调对MCF-7细胞生物学效应的影响。结果发现,乳腺癌组织中EMP1蛋白的相对含量显著低于正常组织(P<0.05)。EMP1蛋白表达水平与T分期、淋巴结转移、临床分期及组织学分级相关(P<0.05)。同时,通过Kaplan-Meier分析发现,EMP1表达缺失与总体生存时间缩短显著相关(P<0.05)。结果显示,与未转染EMP1的MCF-7细胞相比,转染EMP1的MCF-7细胞存活率更低、细胞凋亡率更高、迁移和侵袭能力显著下降、半胱天冬酶-9(caspase-9)水平更高,血管内皮生长因子C(VEGFC)蛋白表达更低(P<0.05)。乳腺癌中EMP1表达降低,且与淋巴结转移、临床分期、组织学分级、总体生存时间缩短及T分期显著相关,提示EMP1可能通过调节caspase 9和VEGFC蛋白的表达,作为乳腺癌MCF-7细胞的负调节因子发挥重要作用。