CAJ | 학술논문

目的探讨肝再生磷酸酶-3（ PRL-3）对人肺癌细胞侵袭能力的影响及相关机制。方法采用RT-PCR和Western blot检测高低转移能力不同的4种肺癌细胞及HBE细胞PRL-3 mRNA及蛋白的表达。免疫荧光检测A549细胞、 HBE细胞中PRL-3的定位。 PRL-3特异性小干扰RNA（ siRNA）转染高转移肺癌细胞株A549后，采用RT-PCR和Western blot检测肺癌细胞PRL-3和mDia1 mRNA及蛋白表达，MTT、迁移试验和Transwell检测肺癌细胞系增殖及迁移侵袭能力， G-LISA 方法检测RhoA 活性。结果肺癌细胞系（ A549，CI-H661） PRL-3 mRNA及蛋白表达高于HBE细胞。 A549细胞系中PRL-3定位于细胞质，而HBE细胞系中PRL-3则定位于细胞核周围。 PRL-3 siRNA能够显著下调A549细胞PRL-3 mRNA和蛋白表达水平，引起RhoA活性降低及其下游作用物 mDia1的表达降低， A549细胞的迁移侵袭能力降低（ P ＜0.01）。结论 PRL-3的表达与肺癌细胞系的迁移侵袭能力有关，PRL-3可能通过调控RhoA活性及mDia1表达促进肺癌细胞迁移侵袭。
목적탐토간재생린산매-3（ PRL-3）대인폐암세포침습능력적영향급상관궤제。방법채용RT-PCR화Western blot검측고저전이능력불동적4충폐암세포급HBE세포PRL-3 mRNA급단백적표체。면역형광검측A549세포、 HBE세포중PRL-3적정위。 PRL-3특이성소간우RNA（ siRNA）전염고전이폐암세포주A549후，채용RT-PCR화Western blot검측폐암세포PRL-3화mDia1 mRNA급단백표체，MTT、천이시험화Transwell검측폐암세포계증식급천이침습능력， G-LISA 방법검측RhoA 활성。결과폐암세포계（ A549，CI-H661） PRL-3 mRNA급단백표체고우HBE세포。 A549세포계중PRL-3정위우세포질，이HBE세포계중PRL-3칙정위우세포핵주위。 PRL-3 siRNA능구현저하조A549세포PRL-3 mRNA화단백표체수평，인기RhoA활성강저급기하유작용물 mDia1적표체강저， A549세포적천이침습능력강저（ P ＜0.01）。결론 PRL-3적표체여폐암세포계적천이침습능력유관，PRL-3가능통과조공RhoA활성급mDia1표체촉진폐암세포천이침습。
Objective To research the affects of endogenous phosphatase of regenerating liver-3 ( PRL-3 ) on the migration and invasion of epithelial cells .Investigate the substrates or pathways that PRL-3 interacts with. Methods PRL-3 mRNA and protein expression was examined by RT-PCR and Western blot analysis in human bronchial epithelial cells and in 4 human lung cancer cell lines .The specificity small interfering RNA against PRL-3 were transfected into highly metastatic lung cancer cell line A 549,then the change of mRNA and protein levels of PRL-3 were observed.The change of RhoA activity (G-LISA experiment) and the protein level of mDia1,which is the downstream targets of RhoA were also detected .Results All four lung cancer cell lines expressed relatively high levels of PRL-3 mRNA and protein,especially the A549 and NCI-H661 cell lines.The HBE cell line had low levels of PRL-3 mRNA and protein .In A549 cells PRL-3 localized to discrete structures in the cytoplasm , but in Human Bronchial Epithelial Cells PRL-3 localized around the nucleus .A549 cells transfected with PRL-3 siRNA had decreased levels of PRL-3 mRNA,protein and a reduced ability to invade through Matrigel (P<0.01).Decreased of RhoA activity and protein level of its downstream targets mDia 1 were detected at the same time .Conclusion The expression of PRL-3 is related with the migration and invasion abilities of lung cancer cell line A 549,PRL-3 may regulate RhoA activity and mDia1 to promote cell migration and invasion in lung cancer cell .