慢病毒介导CIB1表达下调对乳腺癌细胞生物学行为的影响
Effects of Lentivirus-Mediated CIB1 Silencing on Biological Behavior of Breast Cancer Cells
贾庆华 1王晓宇 2张富婷 2马俊 2牛廷献2
作者信息
- 1. 730050 兰州,联勤保障部队第九四〇医院检验科;730050 兰州,甘肃省干细胞与基因药物重点实验室
- 2. 730050 兰州,甘肃省干细胞与基因药物重点实验室;730050 兰州,联勤保障部队第九四〇医院基础医学实验室
- 折叠
摘要
目的 探讨CIB1对三阴性乳腺癌细胞增殖、迁移、凋亡和侵袭的影响及作用机制.方法 将MDA-MB-231和MDA-MB-468细胞分为CIB1敲低组(感染CRISPR/Cas9慢病毒)和阴性对照组.采用CCK-8和EdU实验检测各组细胞增殖,流式细胞术测定细胞凋亡,划痕实验和Transwell实验检测细胞迁移及侵袭.采用qRT-PCR和Western blot检测细胞内β-连环蛋白(β-Catenin)、活化蛋白C(APC)、糖原合酶激酶3β(GSK-3β)、c-myc的mRNA和蛋白表达水平.结果 与阴性对照组比较,CIB1敲低组增殖、侵袭和迁移细胞数减少(P<0.05),凋亡细胞数增加(P<0.05),β-Catenin、APC、c-myc的mRNA和蛋白表达量降低(P<0.05),GSK-3β的mRNA和蛋白表达量增高(P<0.05).结论 敲低CIB1可抑制乳腺癌细胞增殖、迁移及侵袭,促进细胞凋亡,其机制可能与抑制Wnt/β-Catenin信号通路有关.
Abstract
Objective To investigate the effects of calcium and integrin-binding protein 1(CIB1)on the cell proliferation,invasion,apoptosis,and migration of triple-negative breast cancer cells and its possible mechanism.Methods MDA-MB-231 and MDA-MB-468 cells were divided into CIB1-knockdown(infected with CRISPR/Cas9 lentivirus)and negative-control groups.Cell Counting Kit-8(CCK-8)and 5-ethynyl-2′-deoxyuridine(EdU)assays were performed to detect cell proliferation.Cell apoptosis was determined through flow cytometry.Scratch and Transwell experiments were conducted to measure the migration and invasion abilities of cells.The mRNA and protein expression levels of β-catenin,adenomatous polyposis coli(APC),glycogen synthase kinase 3β(GSK-3β),and c-myc were detected via real-time quantitative polymerase chain reaction and Western blot.Results Compared with the negative-control group,the CIB1-knockdown group showed decreased cell proliferation,invasion,and migration(P<0.05)and increased cell apoptosis(P<0.05).The mRNA and protein expressions of β-catenin,APC,and c-myc decreased(P<0.05),and that of GSK-3β increased(P<0.05).Conclusion CIB1 knockdown can inhibit cell proliferation,invasion,and migration and promote the apoptosis of breast cancer cells.Its mechanism may be related to the inhibition of Wnt/β-catenin signaling pathway.
关键词
CIB1/三阴性乳腺癌/增殖/迁移/侵袭/Wnt/β-catenin信号通路Key words
CIB1/Triple negative breast cancer/Proliferation/Migration/Invasion/Wnt/β-catenin signaling pathway引用本文复制引用
基金项目
联勤保障部队第九四〇医院实验室培育项目(2021yxky083)
出版年
2024
国家科技期刊平台
NSTL
万方数据