摘要
目的探讨miR-125a-5p抑制乳腺癌细胞侵袭与转移的潜在机制。方法采用实时荧光定量PCR检测组织及细胞中miR-125a-5p的表达量;利用瞬时转染技术将过表达质粒转染到MDA231中,Transwell侵袭实验检测miR-125a-5p对各组细胞侵袭的影响;Western blot检测各组细胞Akt的磷酸化情况,以及基质金属蛋白酶2(matrix metalloproteinase 2,MMP-2)、MMP-9的表达量。结果通过临床标本的相关检测发现,miR-125a-5p在乳腺癌组织中的表达量明显低于癌旁相对正常组织,并且与淋巴结转移等有关。体外细胞培养实验发现,过表达miR-125a-5p细胞组的侵袭能力明显降低,Akt的磷酸化明显减弱,并且MMP-2和MMP-9表达量下降;而同时过表达GRB相关蛋白2(GRB-associated binding protein2,GAB2)和miR-125a-5p细胞组的侵袭能力明显增强,Akt的磷酸化明显增强,并且MMP-2和MMP-9表达量明显升高,揭示了GAB2和miR-125a-5p共同通过PI3K/Akt通路影响MMP-2和MMP-9的表达。结论 miR-125a-5p能够通过PI3K/Akt/MMP信号通路,抑制乳腺癌细胞的侵袭与转移。
Aim To investigate the potential mechanism of miR-125 a-5 p inhibiting invasion and metastasis of breast cancer cells. Methods qRT-PCR was used to detect the expression of miR-125 a-5 p in tissues and cells. The transient transfection technique was used to transfect the overexpressed plasmid into MDA231. Transwell invasion assay was used to detect the effect of miR-125 a-5 p on invasion ability of breast cancer cells in each group. Western blot was used to detect the phosphorylation level of Akt,the expression of matrix metalloproteinase 2（ MMP-2） and matrix metalloproteinase 9（ MMP-9） in each group of breast cancer cells. Results Results of clinical specimens detection showed that the expression of miR-125 a-5 p in breast cancer tissues was significantly lower than that in adjacent normal tissues, which was related with lymph node metastasis. In vitro cell culture experiment showed that the invasion ability of the cells transfected with miR-125 a-5 p was significantly reduced,the phosphorylation of Akt significantly decreased,and the expression of MMP-2 and MMP-9 was down-regulated.At the same time,the invasion of the cells transfected with GRB associated-binding protein 2（ GAB2） and miR-125 a-5 p was significantly enhanced,Akt phosphorylation was boosted obviously,and the expression of MMP-2 and MMP-9 increased significantly. It revealed that GAB2 and miR-125 a-5 p jointly passed through the PI3 K/Akt pathway, thus affecting the changes of MMP-2 and MMP-9. Conclusion miR-125 a-5 p inhibits the invasion and metastasis of breast cancer through PI3 K/Akt/MMP signal pathway.
作者
杨玉玲
王照岩
杨志一
李洪利
尹崇高
刘雨清
YANG Yu-ling;WANG Zhao-yan;YANG Zhi-yi;LI Hong-li;YIN Chong-gao;LIU Yu-qing(Dept of Pathology,College of Clinical Medicine;Medical Research Center;College of Nursing,Weifang Medical University,Weifang Shandong 261053,China)
出处
《中国药理学通报》
CAS
CSCD
北大核心
2018年第8期1145-1149,共5页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 81402389
81641111
81702932)
山东省自然科学基金资助项目(No ZR2014HL077
ZR2015HL065)
山东省高等学校科技计划资助项目(No J12LK03
J13LK03)
