中华肿瘤防治杂志2024，Vol.31Issue(14) ：856-864,886.DOI:10.16073/j.cnki.cjcpt.2024.14.02

CIB1通过Wnt/β-catenin信号通路调控三阴性乳腺癌细胞增殖和迁移及侵袭机制研究

Regulation of CIB1 on the proliferation,migration and invasion of triple negative breast cancer cells through Wnt/β-catenin signaling pathway

贾庆华 王晓宇 牛廷献 张富婷 马俊
中华肿瘤防治杂志2024，Vol.31Issue(14) ：856-864,886.DOI:10.16073/j.cnki.cjcpt.2024.14.02
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CIB1通过Wnt/β-catenin信号通路调控三阴性乳腺癌细胞增殖和迁移及侵袭机制研究

Regulation of CIB1 on the proliferation,migration and invasion of triple negative breast cancer cells through Wnt/β-catenin signaling pathway

贾庆华 1王晓宇 2牛廷献 2张富婷 2马俊2
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作者信息

  • 1. 中国人民解放军联勤保障部队第九四○医院检验科,甘肃兰州 730050;甘肃省干细胞与基因药物重点实验室,甘肃兰州730050
  • 2. 中国人民解放军联勤保障部队第九四○医院基础医学实验室,甘肃兰州 730050;甘肃省干细胞与基因药物重点实验室,甘肃兰州730050
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摘要

目的 探讨CIB1对三阴性乳腺癌MDA-MB-231和MDA-MB-468细胞增殖、迁移和侵袭的影响及作用机制.方法 通过慢病毒转染构建过表达CIB1的MDA-MB-231和MDA-MB-468细胞,实验分为NC组和CIB1组.采用细胞计数试剂盒8(CCK8)、5-乙炔基-2'脱氧尿嘧啶核苷(EdU)实验检测细胞增殖活性,流式细胞术检测细胞凋亡情况,划痕实验和Transwell实验检测细胞迁移及侵袭能力.同时,采用实时荧光定量-聚合酶链式反应(qRT-PCR)和蛋白质印迹实验检测细胞内β-catenin、活化蛋白C(APC)、糖原合酶激酶3β(GSK-3β)、c-myc的mRNA和蛋白表达情况.结果 MDA-MB-231 细胞内 NC 组和 CIB1 组 CIB1 蛋白表达水平分别为 0.75±0.17 和 1.53±0.38,t=3.267,P=0.031.MDA-MB-468 细胞内 NC 组和 CIB1 组 CIB1 蛋白表达水平分别为 0.91±0.01 和 1.12±0.07,t=4.953,P=0.008.CCK8实验结果显示,MDA-MB-231细胞中NC组和CIB1组增殖力差异有统计学意义,F组别=120.088,P组别＜0.001;F时间=408.784,P时间＜0.001;F组别×时间=14.177,P组别×时间＜0.001.MDA-MB-468细胞中NC组和CIB1组增殖力差异有统计学意义,F组别=277.649,P组别＜0.001;F时间=1 281.882,P时间＜0.001;F组别×时间=33.378,P组别×时间＜0.001.EdU实验结果显示,MDA-MB-231细胞中NC组和CIB1组增殖率分别为(31.42±0.01)％和(46.20±0.02)％,t=14.610,P＜0.001;MDA-MB-468 细胞中 NC 组和 CIB1 组增殖率分别为(30.57±0.02)％和(42.61±0.03)％,t=6.397,P=0.003.MDA-MB-231 细胞中 NC 组和 CIB1 组细胞凋亡率分别为(4.77±0.01)％和(2.07±0.10)％,t=3.785,P=0.019;MDA-MB-468 细胞中 NC 组和 CIB1 组细胞凋亡率分别为(9.53±0.00)％、(3.90±0.00)％,t=19.390,P＜0.001.MDA-MB-231 细胞中 NC 组和 CIB1 组细胞迁移率分别为(46.20±0.01)％和(64.35±0.10)％,t=3.386,P=0.028;MDA-MB-468 细胞中 NC 组和 CIB1 组细胞迁移率分别为(24.84±0.05)％和(56.83±0.06)％,t=7.278,P=0.002.MDA-MB-231 细胞中 NC 组和 CIB1 组穿膜细胞数分别为(300.67±25.17)和(411.67±24.99)个,t=5.421,P=0.006;MDA-MB-468 细胞中 NC 组和 CIB1 组穿膜细胞数分别为(257.00±31.43)和(388.00±8.72)个,t=6.956,P=0.002.qRT-PCR 和蛋白质印迹实验结果显示,MDA-MB-231 和 MDA-MB-468 细胞中 CIB1 组 β-catenin、APC、c-myc mRNA和蛋白表达均升高,GSK-3β mRNA和蛋白表达均降低,差异均有统计学意义,均P＜0.05.结论 CIB1促进乳腺癌细胞增殖、侵袭和迁移能力,抑制细胞凋亡,可能通过促进Wnt/β-catenin信号通路的激活发挥促癌作用.

Abstract

Objective To investigate the effect and mechanism of CIB1 on the proliferation,migration and invasion of MDA-MB-231 and MDA-MB-468 cells in triple negative breast cancer.Method MDA-MB-231 and MDA-MB-468 cells overexpressing CIB1 were constructed by lentiviral transfection,and the experiment was divided into NC group and CIB1 group.Cell pro-liferation activity was detected using cell count kit 8(CCK8)and 5-ethynyl-2'deoxyuridine nucleoside(EdU)assay,flow cytometry was used to detect cell apoptosis,and scratch and trans well assays was used to detect cell migration and invasion ability.Real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)and western blotting experiments were used to detect mRNA and protein expression of intracellular β-catenin,activated protein C(APC),and glycogen synthase kinase 3 β(GSK-3 β)and c-myc.Results The expression levels of CIB1 protein in NC group and CIB1 group of MDA-MB-231 cells were 0.75±0.17 and 1.53±0.38,t=3.267,P=0.031.The expression levels of CIB1 protein in NC group and CIB1 group of MDA-MB-468 cells were 0.91±0.01 and 1.12±0.07,t=4.953,P=0.008.The CCK8 experiment re-sults showed that there was a statistically significant difference in proliferation ability between NC group and CIB1 group in MDA-MB-231 cells,Fgroup=120.088,Pgroup＜0.001;Ftime=408.784,Ptime＜0.001;Fgrcup×time=14.177,Pgroup×time＜0.001.There was a statistically significant difference in proliferation ability between NC group and CIB1 group in MDA-MB-468 cells,Fgroup=277.649,Pgroup＜0.001;Ftime=1 281.882,Ptime＜0.001;Fgroup×time=33.378,Pgroup×time＜0.001.The EdU experiment results showed that,the proliferation rates of NC group and CIB1 group in MDA-MB-231 cells were(31.42±0.01)％and(46.20±0.02)％,t=14.610,P＜0.001;The proliferation rates of NC group and CIB1 group in MDA-MB-468 cells were(30.57±0.02)％and(42.61±0.03)％,t=6.397,P=0.003.The apoptosis rates of NC group and CIB1 group in MDA-MB-231 cells were(4.77±0.01)％and(2.07±0.10)％,t=3.785,P=0.019;The apoptosis rates of NC group and CIB1 group in MDA-MB-468 cells were(9.53±0.00)％and(3.90±0.00)％,t=19.390,P＜0.001.The cell migration rates of NC group and CIB1 group in MDA-MB-231 cells were(46.20±0.01)％and(64.35±0.10)％,t=3.386,P=0.028;The migration rates of NC group and CIB1 group in MDA-MB-468 cells were(24.84±0.05)％and(56.83±0.06)％,t=7.278,P=0.002.The number of transmembrane penetrating cells in NC group and CIB1 group of MDA-MB-231 cells were 300.67±25.17 and 411.67±24.99,t=5.421,P=0.006;The number of trans-membrane penetrating cells in NC group and CIB1 group of MDA-MB-468 cells were 257.00±31.43 and 388.00±8.72,t=6.956,P=0.002.qRT-PCR and western blot experiments showed that the expressions of β-catenin,APC,c-myc mR-NA and protein in CIB1 group were increased,mRNA and protein expressions of GSK-3β were decreased in MDA-MB-231 and MDA-MB-468 cells,all P＜0.05.Conclusions CIB1 may promote the proliferation,invasion and migration of breast cancer cells,and inhibit the apoptosis of breast cancer cells through Wnt/β-catenin signaling pathway to promote breast cancer progression.

关键词

钙整合素结合蛋白1/三阴性乳腺癌/增殖/迁移/侵袭/Wnt/β-catenin信号通路

Key words

calcium and integrin-binding protein 1/triple negative breast cancer/proliferation/migration/invasion/Wnt/β-catenin signaling pathway

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基金项目

联勤保障部队第九四○医院实验室培育项目(2021yxky083)

出版年

2024
中华肿瘤防治杂志
中华预防医学会 山东省肿瘤防治研究院

中华肿瘤防治杂志

CSTPCD北大核心
影响因子：1.292
ISSN：1673-5269
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