摘要
目的研究细胞角蛋白18(cytokeratin 18,CK18)的异常表达对乳腺癌转移的影响,并探寻其作用靶点以阐明机制。方法CK18在人乳腺癌组织和细胞中的表达分别用免疫组织化学染色法、Western blot法测定;shRNA法有效敲低人乳腺癌MCF-7细胞的CK18表达,用划痕实验进一步确定CK18对乳腺癌细胞转移的影响;CK18和非肌肉肌球蛋白ⅡA(non-muscle myosinⅡA,NMIIA)在乳腺癌细胞中的共定位通过免疫荧光细胞化学双重染色法确定;Western blot法定量检测人乳腺癌细胞中NMIIA和c-Abl-ERK通路蛋白水平。结果CK18在有淋巴结转移的乳腺癌组织的表达少于原位乳腺癌、在高侵袭性MDA-MB-231的表达少于MCF-7细胞;敲低CK18增强MCF-7细胞划痕72 h的转移能力;CK18和NMIIA共定位表达于乳腺癌细胞质;敲低CK18可上调MCF-7细胞的NMIIA表达,并激活其c-Abl-ERK通路。结论CK18下调促进乳腺癌转移,其作用可能与增加NMIIA表达、激活c-Abl-ERK通路有关。
Aim To investigate the role of aberrant cytokeratin 18(CK18)expression in breast cancer metastasis,and to elucidate the mechanism by identifying its target.Methods The expression of CK18 in human breast cancer tissues and cells was determined using immunohistochemical staining and Western blot,respectively.CK18 expression in human breast cancer MCF-7 cells was effectively down-regulated by shRNA,and its effect on breast cancer metastasis was further determined by scratch wound healing assay.The co-location of CK18 and non-muscleⅡA(NMIIA)in MCF-7 cells was examined using double immunofluorescence staining.The effect of CK18 down-regulation on the levels of NMIIA and c-Abl-ERK signaling was quantified by Western blot.Results Lower CK18 levels was found in metastatic than that in primary breast cancer tissues and in highly invasive MDA-MB-231 than that in MCF-7 cells.CK18 down-regulation promoted the wound repair ability of MCF-7 cells 72h after scratch.CK18 and NMIIA were shown to co-locate in cytoplasm of MCF-7 cells.Moreover,down-regulation of CK18 increased NMIIA expression and activated the c-Abl-ERK signaling pathway in MCF-7 cells.Conclusions Down-regulation of CK18 could promote metastasis of breast cancer,which is related to increased NMIIA expression and the activation of c-Abl-ERK signaling pathway.
作者
师锐赞
牛亚楠
高宇
李克敏
李瑞涵
刘临红
陈敏
张轩萍
SHI Rui-zan;NIU Ya-nan;GAO Yu;LI Ke-min;LI Rui-han;LIU Lin-hong;CHEN min;ZHANG Xuan-ping(Dept of Pharmacology, Shanxi Medical University, Yuci Shanxi 030600, China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2022年第6期861-865,共5页
Chinese Pharmacological Bulletin
基金
山西省自然科学基金资助项目(No 201901D111200)
国家自然科学基金资助项目(No 81502641)
大学生创新创业训练计划项目(No 202110114026)。
