吗啡促乳腺癌细胞转移的YAP相关信号机制研究

目的 探讨YAP相关的分子信号通路在吗啡促乳腺癌细胞转移中的作用及机制.方法 应用RNA干扰技术降低人乳腺癌BT474细胞中YAP蛋白的表达,体外Transwell细胞转移实验验证YAP蛋白被抑制后,吗啡对乳腺癌细胞转移的刺激作用.采用Western blot技术检测吗啡处理后,乳腺癌细胞中YAP蛋白及磷酸化YAP蛋白的表达;细胞免疫荧光分析吗啡对YAP蛋白细胞内定位的影响.结果 siRNA能有效抑制乳腺癌BT474细胞中YAP蛋白的表达,吗啡干预后的乳腺癌BT474细胞转移数显著高于对照组（79.4±5.48 vs.32.2±6.42,P=0.000 5）,而当YAP蛋白被抑制后,吗啡未能增...

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Bibliographic Details
Published in西安交通大学学报（医学版） Vol. 35; no. 5; pp. 618 - 621
Main Author 杨远东 张灵敏 袁慧 袁伟
Format Journal Article
LanguageChinese
Published 西安交通大学医学院第一附属医院麻醉科,陕西西安,710061 2014
Subjects
YAP蛋白
乳腺癌
吗啡
转移
吗啡
乳腺癌
yes-associated protein (YAP)
migration
breast cancer
转移
YAP蛋白
morphine
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ISSN1671-8259
DOI10.7652/jdyxb201405010

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Summary:目的 探讨YAP相关的分子信号通路在吗啡促乳腺癌细胞转移中的作用及机制.方法 应用RNA干扰技术降低人乳腺癌BT474细胞中YAP蛋白的表达,体外Transwell细胞转移实验验证YAP蛋白被抑制后,吗啡对乳腺癌细胞转移的刺激作用.采用Western blot技术检测吗啡处理后,乳腺癌细胞中YAP蛋白及磷酸化YAP蛋白的表达;细胞免疫荧光分析吗啡对YAP蛋白细胞内定位的影响.结果 siRNA能有效抑制乳腺癌BT474细胞中YAP蛋白的表达,吗啡干预后的乳腺癌BT474细胞转移数显著高于对照组（79.4±5.48 vs.32.2±6.42,P=0.000 5）,而当YAP蛋白被抑制后,吗啡未能增加BT474细胞的转移（29.2±4.08 vs.21.2±2.58,P=0.135 9）;Western blot检测发现,在BT474细胞的培养基中加入吗啡干预后后,磷酸化的YAP含量随着时间的增加而显著降低,但总YAP蛋白的表达无显著变化,同时,细胞免疫荧光证实吗啡能促进BT474细胞中YAP蛋白在细胞核内聚集.结论 吗啡能通过激活细胞中的YAP蛋白从而促进乳腺癌的转移.
Bibliography:Objective To investigate the role of yes-associated protein （YAP） signaling pathway in morphineinduced migration of breast cancer cells.Methods YAP in BT474 cells was down-regulated using siRNA.Migration assays were conducted using in vitro transwell cell migration assays.Western blots were performed to detect changes of YAP and phosphorylated YAP expressions in breast cancer cells after treatment with morphine.The translocation of YAP in cells was analyzed by immunofluorescence staining.Results The expression of YAP in BT474 cells was significantly inhibited by YAP-siRNA.Morphine could significantly increase the migration of breast cancer cells as compared to that of controls （79.4 ± 5.48 vs.32.2-t-6.42,P=0.0005）.Down-regulated YAP led to inhibition of morphine-induced BT474 cell migration （29.2 ± 4.08 vs.21.2 ± 2.58,P =0.135 9）.In addition,morphine-induced YAP dephosphorylation and nuclear translocation in the time-dependent manner.Conclusion Morphine can promote the metastasis of breast cancer by activatin
ISSN:1671-8259
DOI:10.7652/jdyxb201405010