CAJ | 학술논문

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pcDNA3－TRAF4－DM－TRAF质粒）、对照组（转染pcDNA3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pcDNA3－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。
목적：탐토TRAF4재인유선암세포고표체시대세포증식능력적영향。방법본실험공분량조：실험조（재유선암세포MDA－MB－231중전염pcDNA3－TRAF4－DM－TRAF질립）、대조조（전염pcDNA3공질립），용세포면역형광급면역인적법검측TRAF4적표체급정위，면역인적법검측p70S6K급S6단백린산화，용류식세포술검측각기세포비례，MTT방법검측증식능력。결과 TRAF4재유선암세포MDA－MB－231세포질급세포핵중균유표체，병차기재세포핵중적표체현저저우세포질중적표체（ P＜0．05）。재해세포중전염pcDNA3－TRAF4－DM－TRAF질립후，기세포핵내TRAF4표체현저승고（P＜0．05）， p70S6K급S6단백린산화수평현저증가（P＜0．05，P＜0．01），S기세포비례현저증가（P＜0．01），병차세포증식능력현저상조（P＜0．01）。결론상조TRAF4재유선암세포MDA－MB－231세포핵중적표체，가능통과촉진p70S6K급S6단백린산화이촉진해세포적증식。
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 p70S6K and S6 with westernblot.Flow cytometry was used to detecte cell cycle.MTT 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 S6 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 MTT ( P<0.01) .Conclusions_The expression of TRAF4 in nuclei may play an important role in the cell prolif-eratuion by promoting p70S6K and S6 activation.