摘要
[目的]探究REC8(Meiotic recombination protein 8,REC8)调控PAK1对三阴性乳腺癌(Triple-negative breast cancer,TNBC)转移活性的影响。[方法]收集36例TNBC患者的癌组织及癌旁组织,免疫组化分析REC8在TNBC组织及癌旁组织中阳性表达率。将TNBC MDA-MB-231细胞分为3组(2.5×10^(5)个/m L):阴性对照组(转染100nmol/孔、50 nmol/L阴性对照siRNA)、pc DNA REC8组(转染2μg/孔、1μg/μL pc DNA REC8过表达质粒)、siPAK1组(转染100 nmol/孔、50 nmol/L siPAK1)。通过Ed U染色实验分析MDA-MB-231细胞增殖活性,Transwell实验分析MDA-MB-231细胞的侵袭、迁移能力,蛋白免疫印迹实验分析MDA-MB-231细胞中REC8、PAK1蛋白的表达。[结果]与癌旁组织比较,REC8在TNBC组织中表达降低(P<0.05)。与阴性对照组相比,pc DNA REC8、siPAK1组的MDA-MB-231细胞增殖能力下降(P<0.05);pc DNA REC8、siPAK1组的MDA-MB-231细胞的侵袭、迁移能力下降(P<0.05);pc DNA REC8、siPAK1组的MDA-MB-231细胞PAK1蛋白表达降低(P<0.05)。[结论]REC8在TNBC组织中表达降低。上调TNBC MDA-MB-231细胞的REC8表达能够通过抑制PAK1进而减弱MDA-MB-231细胞的增殖、侵袭、迁移活性。
[Objective]To investigate the effect of REC8 on the metastatic activity of triple-negative breast cancer(TNBC)by regulating PAK1.[Method]The cancer tissues and adjacent tissues of 36 patients with TNBC were collected.The positive expression rate of REC8 in TNBC tissues and adjacent tissues was analyzed by immunohistochemistry.The TNBC MDA-MB-231 cells were divided into 3 groups(2.5×10^(5) cells/mL):the negative control group(transfected with volume of 100 nmol per well of non-functional negative control siRNA at 50 nmol/L),the pcDNA REC8 group(transfected with volume of 2μg per well of pcDNA REC8 overexpression plasmid at concentration of 1μg/μL),and the siPAK1 group(transfected with volume of 100 nmol per well siPAK1 at concentration of 50 nmol/L).The proliferation activity of MDA-MB-231 cells was analyzed by EdU staining.The invasion and migration ability of MDA-MB-231 cells was analyzed by Transwell assay.The expressions of REC8 and PAK1 proteins in MDA-MB-231 cells were analyzed by Western blot.[Result]REC8 expression was significant⁃ly lower in TNBC tissues than in adjacent tissues(P<0.05).Compared with the negative control group,the proliferation ability of MDA-MB-231 cells in the pcDNA REC8 and siPAK1 groups was decreased(P<0.05).The invasion and migration abil⁃ities of MDA-MB-231 cells in the pcDNA REC8 and siPAK1 groups were significantly decreased(P<0.05).The expres⁃sion of PAK1 protein in MDA-MB-231 cells was significantly decreased in the pcDNA REC8 and siPAK1 groups(P<0.05).[Conclusion]The expression of REC8 is decreased in TNBC tissues.Upregulation of REC8 expression in TNBC MDA-MB-231 cells can inhibit PAK1,thereby reducing the proliferation,invasion,and migration activities of MDA-MB-231 cells.
作者
贺芳
徐盼玲
李平
HE Fang;XU Panling;LI Ping(Graduate School of Anhui University of Traditional Chinese Medicine,Hefei 230031;Department of Integrated Traditional Chinese and Western Medicine Oncology,The First Affiliated Hospital of Anhui Medical University,Hefei 230032,China)
出处
《生物技术》
2025年第6期761-765,804,共6页
Biotechnology
基金
安徽省高等院校科学研究项目(2022AH051162)。
