摘要
目的:通过生物信息学分析,利用癌症基因组图谱(TCGA)和高通量基因表达数据库(GEO)研究VI胶原蛋白α3链(COL6A3)对三阴性乳腺癌(TNBC)细胞增殖、迁移、侵袭及穿透血脑屏障能力的影响。方法:通过TCGA和GEO数据库进行生物信息学分析,比较COL6A3在正常乳腺组织、乳腺癌(BC)组织及BC脑转移瘤组织中的表达,并进行基因本体论(GO)和京都基因与基因组百科全书(KEGG)富集分析。使用实时荧光定量逆转录聚合酶链式反应(RT-qPCR)和蛋白质印迹法(WB)检测COL6A3在TNBC细胞与正常乳腺细胞中的表达。将TNBC细胞通过慢病毒感染分为转染阴性对照组(NC组)和敲低组(sh-COL6A3组),并通过RT-qPCR和WB检测转染情况。使用细胞计数的盒子-8(CCK-8)、细胞克隆实验、划痕实验、Transwell实验及体外血脑屏障模型检测细胞的增殖、迁移、侵袭和穿透血脑屏障能力。使用WB验证COL6A3与PI3K/AKT通路的相关性。结果:TCGA数据分析,COL6A3在BC样本中表达水平显著高于正常乳腺样本(P<0.05),且富集于PI3K/AKT通路。GSE125989数据分析显示,COL6A3在BC脑转移瘤样本中表达水平明显高于原发性BC样本(P<0.05);与正常乳腺组织细胞相比,COL6A3在TNBC细胞中的信使核糖核酸(mRNA)和蛋白表达上调(P<0.05);与NC组相比,敲低组TNBC细胞增殖、迁移、侵袭及穿透血脑屏障能力均显著下降(P<0.05),且敲低组PI3K/AKT通路标志蛋白表达下调(P<0.05)。结论:COL6A3通过调控PI3K/AKT通路促进TNBC细胞增殖、迁移、侵袭及穿透血脑屏障。
Objective To investigate the effects of VI collagenα3 chain(COL6A3)on the proliferation,migration,invasion and blood-brain barrier penetration ability of triple-negative breast cancer(TNBC)cells through bioinformatics analysis using the cancer genome atlas(TCGA)and the high-throughput gene expression database(GEO).Methods Bioinformatics analysis was conducted using the TCGA and GEO databases to compare the expression of COL6A3 in normal breast tissues,breast cancer(BC)tissues,and BC brain metastases tissues.Gene ontology(GO)and Kyoto encyclopedia of genes and genomes(KEGG)enrichment analyses were also performed.The expression of COL6A3 in TNBC cells and normal breast cells was detected by real-time fluorescence quantitative reverse transcription polymerase chain reaction(RT-qPCR)and Western blot(WB).TNBC cells were divided into the transfection negative control group(NC group)and the knockdown group(sh-COL6A3 group)by lentivirus infection,and the transfection status was detected by RT-qPCR and WB.The proliferation,migration,invasion and penetration ability of cells through the blood-brain barrier were detected using the cell counting box-8(CCK-8),cell cloning assay,scratch assay,Transwell assay and in vitro blood-brain barrier model.The correlation between COL6A3 and the PI3K/AKT pathway was verified using WB.Results According to the TCGA data analysis,the expression level of COL6A3 in BC samples was significantly higher than that in normal breast samples(P<0.05),and it was enriched in the PI3K/AKT pathway.Data analysis of GSE125989 showed that the expression level of COL6A3 in BC brain metastatic tumor samples was significantly higher than that in primary BC samples(P<0.05);Compared with normal breast tissue cells,the expression of messenger ribonucleic acid(mRNA)and protein of COL6A3 in TNBC cells was upregulated(P<0.05);Compared with the NC group,the proliferation,migration,invasion and blood-brain barrier penetration ability of TNBC cells in the sh-COL6A3 group were significantly decreased(P<0.05),and the expression of PI3K/AKT pathway marker proteins was down-regulated in the sh-COL6A3 group(P<0.05).Conclusion COL6A3 promotes the proliferation,migration,invasion and penetration of TNBC cells into the blood-brain barrier by regulating the PI3K/AKT pathway.
作者
梁柱明
莫雅楠
张玉姣
于燕菁
姬逸男
LIANG Zhuming;MO Yanan;ZHANG Yujiao;YU Yanjing;JI Yinan(Department of Breast Surgery,Guangxi Medical University Affiliated Tumor Hospital/Guangxi University Laboratory of Breast Cancer Diagnosis and Treatment Research,Guangxi Nanning 530021)
出处
《深圳中西医结合杂志》
2025年第10期1-4,14,I0002,I0003,F0003,共8页
Shenzhen Journal of Integrated Traditional Chinese and Western Medicine
基金
国家自然科学基金资助项目(No.81860464)
广西自然科学基金资助项目(2020GXNSFBA297072)。
关键词
三阴性乳腺癌
VI胶原蛋白α3链
脑转移
Triple-negative breast cancer
VI collagenα3 chain
Brain metastasis
