转染pcDNA3-TRAF4-DM-TRAF促进人乳腺癌细胞MDA-MB-231细胞增殖  

Transfecting pcDNA3-TRAF4-DM-TRAF improves proliferation of human breast cancer cell MDA-MB-231

作　　者：张晓丽[1] 米小轶[2] 

机构地区：[1]沈阳市妇婴医院生殖中心,辽宁沈阳110014 [2]中国医科大学附属第一医院病理科,辽宁沈阳110001

出　　处：《基础医学与临床》2015年第10期1382-1386,共5页Basic and Clinical Medicine

摘　　要：目的探讨TRAF4在人乳腺癌细胞高表达时对细胞增殖能力的影响。方法本实验共分两组:实验组(在乳腺癌细胞MDA-MB-231中转染pc DNA3-TRAF4-DM-TRAF质粒)、对照组(转染pc DNA3空质粒),用细胞免疫荧光及免疫印迹法检测TRAF4的表达及定位,免疫印迹法检测p70S6K及S6蛋白磷酸化,用流式细胞术检测各期细胞比例,MTT方法检测增殖能力。结果 TRAF4在乳腺癌细胞MDA-MB-231细胞质及细胞核中均有表达,并且其在细胞核中的表达显著低于细胞质中的表达(P<0.05)。在该细胞中转染pc DNA3-TRAF4-DM-TRAF质粒后,其细胞核内TRAF4表达显著升高(P<0.05),p70S6K及S6蛋白磷酸化水平显著增加(P<0.05,P<0.01),S期细胞比例显著增加(P<0.01),并且细胞增殖能力显著上调(P<0.01)。结论上调TRAF4在乳腺癌细胞MDA-MB-231细胞核中的表达,可能通过促进p70S6K及S6蛋白磷酸化而促进该细胞的增殖。Objective To investigate whether overexpression of TRAF4 in human breast cancer may have impact on its cell proliferation. Methods This study has two groups, MDA-MB-231 transfected with pcDNA3-TRAF4-DM- TRAF or pcDNA3. We detected the expression and localization of TRAF4 with immunofluorescence and western blot. We detected the expression of the phosphorylation level of pTOS6K and $6 with westernblot. Flow cytometry was used to detecte cell cycle. MTr Assay was used to detected cell reproductive capacity. Results TRAF4 local- ized in both cytoplasm and nuclei in MDA-MB-231. Nuclear expression of TRAF4 in nuclei was lower than that in cytoplasm （P 〈 0.05）. After pcDNA3-TRAF4-DM-TRAF was transfected into the cells, the expression of TRAF4 in nuclei was increased （ P 〈 0. 05 ）. The phosphorylation level of p70S6K and $6 significantly increaseced （ P 〈 0. 05 ,P 〈 0.01 ）. More S phase cells were recorded（P 〈 0. 01 ） by FCM. The cell proliferation was promoted by MTr （P 〈 0. 01 ）. Conclusions The expression of TRAF4 in nuclei may play an important role in the cell prolif- eratuion by promoting p70S6K and $6 activation.

关 键 词：乳腺癌 TRAF4 细胞增殖 

分 类 号：R737.9[医药卫生—肿瘤]