摘要
目的三阴性乳腺癌作为乳腺癌的特殊类型,尚缺乏有效的靶向治疗手段。文中旨在观察溶血磷脂酸(LPA)对三阴性乳腺癌的影响,探讨LPA-Hippo-Yes相关蛋白(YAP)信号通路在促进三阴性乳腺癌侵润转移中的作用及机制。方法体外合成YAP序列特异性小干扰RNA,应用质粒转染MDA-MB-231乳腺癌细胞株为实验组,非特异性siRNA转染MDA-MB-231细胞为阳性对照组,未转染MDA-MB-231细胞为空白对照组,每组设2个平行孔。检测各组对Hippo-YAP信号的影响,其作用机制及对Hippo-YAP上、下游信号通路调控。结果实验组总蛋白中YAP含量、细胞转染48h后的侵袭、转移能力[(0.035±0.005)、(2.200±1.000)个、(3.500±0.800)个]较阳性对照组[(0.343±0.012)、(27.600±5.100)个、(22.300±5.000)个]、空白对照组[(0.384±0.017)、(26.500±4.800)个、(22.350±6.000)个]明显降低(P<0.05)。实验组60、120、240 min时YAP表达量较阳性对照组、空白对照组显著降低(P<0.05)。实验组10、20、50μmol/L的YAP相对表达量显著低于阳性对照组、空白对照组(P<0.05)。C3转移酶和Y27632分别干预后,实验组p YAP mRNA含量(0.255±0.052、0.326±0.017)与空白对照组((0.048±0.032、0.534±0.017)、阳性对照组(0.052±0.021、0.528±0.024)比较差异有统计学意义(P<0.05)。实验组YAP m NA、AREG m NA的表达量(0.176±0.032、0.263±0.008)较空白对照组(0.043±0.013、0.051±0.014)、阴性对照组(0.049±0.025、0.057±0.043)显著升高(P<0.05)。结论 LPA诱导乳腺癌细胞侵袭转移,具有YAP蛋白依赖性,并具有时间与浓度依赖的特点;LPA-Hippo-YAP信号传导通路可能是促进三阴性乳腺癌细胞长时间转移的机制之一。
Objective Triple negative breast cancer（TNBC）,a special breast cancer subtype,is lack of effective target therapy. The article aimed to investigate the role of lysophosphatidic acid（LPA） and Hippo Yes-associated protein（Hippo-YAP） signaling pathway in TNBC invasion and metastasis and the mechanisms. Methods The specific small interfering RNA（siRNA） of YAP was synthetized in vitro,and was transfected into MDA-MB-231 cells using liposome transfection. The experiment was divided into YAP-siRNA group,positive control group and blank control group. Each group is provided with 2 parallel holes. Evaluation was made on the effects of each group on Hippo-YAP,the mechanisms and regulation on upstream and downstream molecules of Hippo-YAP pathway.Results In experiment group,YAP content,the capacity of invasion and metastasis after transfection（[0. 035 ± 0. 005],[2. 200 ±1.000],[3.500±0.800]） significantly decreased compared with positive control group（[0.343±0.012],[27.600±5.100],[22.300±5.000]） and blank control group（[0.384±0.017],[26. 500 ± 4. 800],[22. 350 ± 6. 000]）（P 0. 05）. YAP expression levels at 60 min,120 min,and 240 min in experiment group significantly decreased compared with positive control group and blank control group（P〈0.05）. YAP relative expression levels of 10,20,50 μmol/Lwere significantly lower than those of positive control group and blank control group（P〈0.05）. After respective interference of C3 transferase and Y27623,significant difference was found in the p YAP mRNA contents of experiment group（[0.255±0.052],[0.326± 0.017]）,blank control group（[0. 048 ± 0. 032],[0. 534 ± 0. 017]） and positive control group（[0.052±0.021],[0.528±0.024]）（P〈0.05）. The expression levels of YAP m NA and AREG m NA significantly increased in experiment group（[0.176± 0. 032],[0. 263 ± 0. 008]） compared with blank control group（[0. 043 ± 0. 013],[0. 263 ±0.008]） and positive control group（[0.049±0.025],[0.057±0.043]）（P〈0.05）. Conclusion LPA induces breast cancer invasion and metastasis,which is YAP-dependent,time-dependent and concentration-dependent. LPA-Hippo-YAP singaling pathway may be one of the mechanisms promoting delayed metastasis of TNBC.
出处
《医学研究生学报》
CAS
北大核心
2017年第8期829-833,共5页
Journal of Medical Postgraduates
基金
西安交通医学院附属三二〇一医院重点基金项目(3201YK201539)
关键词
乳腺癌
溶血磷脂酸
Yes相关蛋白
信号通路
Breast cancer
Lysophosphatidic acid
Yes-associated protein
Signaling pathway
作者简介
通信作者：王健生,E-mail：wangjshxjtu@gmail.com
