摘要
背景与目的:硒-甲基硒代半胱氨酸(Se-methylselenocysteine,MSC)是一种天然有机硒,已被证实能预防和治疗多种肿瘤,但其抗肿瘤作用的机制尚有待进一步阐明。本研究旨在通过MSC处理乳腺癌MDA-MB-231细胞,研究其对细胞生物学行为和基质金属蛋白酶-2(matrix metalloproteinase-2,MMP-2)表达的影响。方法:用不同浓度的MSC处理MDA-MB-231细胞,镜下观察细胞形态的改变,细胞计数试剂盒(cell counting Kit-8,CCK-8)、流式细胞仪及软琼脂生长实验分别检测其对肿瘤细胞增殖、周期分布和凋亡及恶性表型的影响,逆转录-多聚酶链反应(reverse transcription-polymerase chain reaction,RT-PCR)和Western blot检测MMP-2 mRNA和蛋白水平的表达,酶联免疫吸附实验(enzyme-linked immunosorbent assay,ELISA)检测细胞培养液中MMP-2蛋白浓度。结果:MSC处理的MDA-MB-231细胞增殖受到抑制,出现S期阻滞和细胞凋亡,软琼脂上细胞集落形成数明显减少。MMP-2mRNA和蛋白的表达却表现出不一致性,50μmol/LMSC处理细胞48h和72h后,MMP-2mRNA表达水平分别上调32.2%和47.1%,而MMP-2蛋白表达水平却分别下调42.4%和50.8%,培养液中的MMP-2浓度也分别下调了56.7%和75.2%;100μmol/LMSC处理细胞48和72h后,MMP-2mRNA表达水平分别上调52.6%和61.3%,而MMP-2蛋白表达水平分别下调72.9%和81.4%,培养液中的MMP-2浓度也分别下调了68.5%和80.9%。结论:MSC抑制乳腺癌MDA-MB-231细胞增殖,诱导S期阻滞及细胞凋亡,降低细胞恶性程度。MSC上调该细胞MMP-2 mRNA的表达,但下调其蛋白的表达和分泌。
BACKGROUND ＆ OBJECTIVE： Se-methylselenocysteine （MSC）, a natural organoselenium compound, has functions on chemoprevention and treatment of many tumors, but the mechanism remains unclear. This study was to investigate the effects of MSC on the biological behavior of and matrix metalloproteinase-2 （MMP-2） expression in human breast cancer MDA-MB-231 cells. METHODS： After treatment of MSC, the proliferation and apoptosis of MDA-MB-231 cells were detected with light microscope and Cell Counting Kit-8 （CCK-8）, cell cycle and apoptosis were determined by flow cytometry, malignant phenotype was determined by soft agarose growth assay, and the expression of MMP-2 was determined by reverse transcription-polymerase chain reaction （RT-PCR） and Western blot. The concentration of MMP-2 protein in culture supernatant was measured with enzyme-linked immunosorbent assay （ELISA）. RESULTS： MSC induced S phase arrest and apoptosis, inhibited the proliferation and colony formation of MDA-MB-231 cells. When treated with 50 μmol/L MSC for 48 h and 72 h, the mRNA levels of MMP-2 were up-regulated by 32.2% and 47.1%, whereas its protein levels were down-regulated by 42.4% and 50.8%, its concentrations in culture supernatant were down-regulated by 56.7% and 75.2%. When treated with 100 μmol/L MSC for 48 h and 72 h, the mRNA levels of MMP-2 were up-regulated by 52.6% and 61.3%, whereas its protein levels were down-regulated by 72.9% and 81.4%, and its concentrations in culture supernatant were down-regulated by 68.5% and 80.9%. CONCLUSIONS： MSC could inhibit proliferation, and induce apoptosis and S phase arrest in MDA-MB-231 cells. MSC could also up-regulate the mRNA level of MMP-2 while down-regulate its protein level and secretion in the cells.
出处
《癌症》
SCIE
CAS
CSCD
北大核心
2008年第2期119-125,共7页
Chinese Journal of Cancer
