摘要
目的研究花生四烯酸(AA)对鼻咽癌细胞增殖的影响及其潜在的分子机制。方法采用CCK-8法检测AA对鼻咽癌细胞系C666-1增殖的影响,以及雷帕霉素(rapamycin,Rap)、PP242与AA共同作用对C666-1细胞增殖的影响。同时采用免疫印迹法检测AA对鼻咽癌C666-1细胞哺乳动物雷帕霉素靶蛋白(mTOR)信号通路活性的影响,包括下游相关蛋白S6、Akt的表达水平及磷酸化水平,以及mTOR通路抑制剂Rap、PP242与AA共同作用对mTOR信号通路的影响。结果 AA可强烈促进C666-1细胞的增殖,mTOR通路抑制剂Rap、PP242均可抑制AA促进C666-1细胞增殖。AA可激活C666-1细胞mTORC1/2下游相关蛋白S6、Akt,使其磷酸化水平升高。mTORC1通路抑制剂Rap可阻断AA对mTORC1的激活,从而降低S6(S235/236)的磷酸化水平,mTORC1/2通路抑制剂PP242可阻断AA对mTORC1/2的激活,从而降低S6(S235/236)和Akt(S473)的磷酸化水平。结论花生四烯酸通过激活mTORC1/2信号通路促进鼻咽癌C666-1细胞系的增殖。
Objective To investigate the mechanism and effect of arachidonic acid （AA） on the proliferation of C666-1 cells. Methods C666-1 cells were treated with AA, Rap, and PP242 in 96-well plates. The proliferation of C666-1 was determined using the CCK-8 cell proliferation assay kit. Western-blot analysis was used to detect the change of the mTOR signaling. Results AA was found to strongly stimulate C666-1 cell proliferation. AA-stimulated proliferation could be inhibited by mTOR pathway inhibitors Rap and PP242.AA was also found to activate the mTORC1/2 downstream signaling proteins $6 and Akt. Furthermore, the activation mTORC1 could be blocked by the mTORC1 pathway inhibitor Rap. The level of phosphorylation of $6（$235/236） was also reduced, mTORC1/2 pathway inhibitor PP＇242 was found to block AA-induced activation of the mTORC1/2 signaling pathway and reduce the phosphorylation level of both $6 （$235/236） and Akt（S473）. Conclusion Arachidonic acid stimulates C666-1 cell proliferation through the activation of mTORC1/2 signaling pathway.
出处
《热带医学杂志》
CAS
2013年第5期525-527,561,共4页
Journal of Tropical Medicine
基金
国家自然科学基金(91029727)
广东省自然科学基金(S2012010008209)
关键词
鼻咽癌
花生四烯酸
MTOR
nasopharyngeal carcinoma
arachidonic acid
mTOR
