cPLA2α在乳腺癌细胞运动侵袭中作用的研究
he Study for the Roles of cPLA2α in Breast Cancer Cells Chemotaxis and Invasion
目的：探讨降低cPLA2α的表达对乳腺癌细胞运动迁移和侵袭能力的影响，并阐述其作用机制。方法：采用免疫组化的方法检测cPLA2α在乳腺癌组织及不同乳腺癌细胞系中的表达情况。应用MTT、趋化和Transwell的方法检测cPLA2α降表达对MDA-MB-231细胞的增殖、迁移和侵袭的影响。应用Western blotting技术检测cPLA2α降表达后，MDA-MB-231细胞在EGF刺激不同时间后Akt（Ser473和Thr308）、integrinβ1和FAK的磷酸化水平变化。通过花生四烯酸的应用探究其对cPLA2α降表达MDA-MB-231细胞的趋化、迁移及FAK磷酸化水平的影响。应用免疫缺陷鼠（SCID鼠）的动物模型观察cPLA2α在乳腺癌细胞转移中的作用。结果：cPLA2α在乳腺癌组织中的表达率显著高于相应的癌旁组织；cPLA2α降表达后的MDA-MB-231细胞在增殖、迁移和侵袭能力上显著降低，其Akt（Ser473和Thr308）、integrinβ?和FAK的磷酸化水平表达与对照组相比明显降低；花生四烯酸可以恢复cPLA2α降表达的MDA-MB-231细胞的划痕和趋化运动能力以及FAK磷酸化表达水平；cPLA2α表达降低的乳腺癌MDA-MB-231细胞注入SCID鼠体内，肿瘤细胞的自发转移和被动转移能力降低。结论：cPLA2α参与EGF诱导的乳腺癌细胞的运动和侵袭；cPLA2α是通过调控Akt、integrinβ1和FAK的磷酸化水平调节乳腺癌细胞的趋化运动；花生四烯酸及其代谢产物与cPLA2α降表达乳腺癌细胞的运动能力变化有密切关系；cPLA2α参与体内乳腺癌细胞的转移。
Objectives: To investigate the role of cPLA2α in breast cancer cell invasion and illustrate the mechanisms of cPLA2α signaling pathway in breast cancer cell invasion. Method: Breast cancer cells were transfected with small RNA interference plasmids to disrupt cPLA2α expression. cPLA2α protein levels and mRNA levels were detected by Western blotting and RT-PCR. Scratch assay and chemotaxis assay were examined to detect the migration and chemotaxis ability of breast cancer cells. The in vitro invasion ability of breast cancer cells was examined using matrigel invasion assay. Western blotting was used to check the level of phosphorylated Akt, integrinβ1?and FAK, and check the rescue expression level of phosphorylated FAK by adding arachidonic acid to sicPLA2α/MDA231 cell, and meanwhile detect the migration and chemotaxis ability of the cells. The sicPLA2α/MDA231 cell was injected or hypodermic inoculated into SCID rat in order to examine the in vivo invasion ability of breast cancer cells. Result: The cPLA2α-reduced MDA231 cells showed decreased chemotaxis ability compared with the control cells. The invasion assay showed prominent differences between the cPLA2α-reduced MDA231 cells and the control cells. In the cPLA2α-reduced MDA231 cells, the actin polymerization in response to the EGF stimulation was significantly reduced. Western blotting results showed that the level of polymerization Akt, polymerization integrinβ??and polymerization FAK was inhibited for the cPLA2α reduction. The invasion and metastasis assay of human breast cancer cells in the SCID rat showed that the number of tumors in the surface of lung has been clearly decreased in the cPLA2α-impaired MDA231 cells group. Conclusion: cPLA2α participated in EGF-induced chemotaxis and invasion of human breast cancer cells, cPLA2α modulate the EGF-induced expression of phosphorylated Akt, integrinβ??and FAK, which regulated chemotaxis and invasion; Arachidonic acid and its metabolite were associated with chemotaxis and migration of breast cancer cells; cPLA2α plays an important role in invasion and metastasis of human breast cancer cells in vivo.
陈丽维、付慧、郭华、陈璐、张宁
肿瘤学基础医学分子生物学
乳腺癌胞浆型磷脂酶A2α（cPLA2α）侵袭趋化转移
breast cancercytosolic phospholipase A2αinvasionchemotaxismetastasis
陈丽维,付慧,郭华,陈璐,张宁.cPLA2α在乳腺癌细胞运动侵袭中作用的研究[EB/OL].(2015-11-20)[2025-08-11].http://www.paper.edu.cn/releasepaper/content/201511-330.点此复制
评论