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S6K1调控的蛋白表达图谱及其对乳腺癌细胞恶性表型的影响OACSTPCD

Expression of ribosomal protein S6K1 and its regulation on malignant phenotypes in breast cancer cells

中文摘要英文摘要

目的:探究核糖体蛋白S6激酶β-1(S6K1)所调控的蛋白表达图谱及其对乳腺癌细胞恶性表型的影响.方法:通过cBioPortal和GEPIA数据库分析S6K1基因在乳腺癌组织中的扩增及表达,并评估其表达对乳腺癌患者临床特征的影响;利用小干扰RNA在乳腺癌MCF7细胞中敲降S6K1,通过质谱技术检测S6K1所调控的蛋白表达图谱;采用细胞成球实验探究S6K1表达水平改变对乳腺癌细胞干性的影响;通过细胞迁移实验探究S6K1对乳腺癌细胞迁移能力的影响.结果:S6K1基因在乳腺癌组织中存在高频扩增现象,在浸润性乳腺癌中的扩增率高于非浸润性乳腺癌(P<0.05),且S6K1的高频扩增与患者的不良预后相关(P<0.01);S6K1在乳腺癌组织中的表达水平较癌旁组织高(P<0.01),而S6K1在乳腺癌不同临床分期中表达水平无明显差异(P>0.05).质谱检测表明敲降S6K1后,乳腺癌MCF7细胞中有251个蛋白表达上调,224个蛋白表达下调;GO通路富集分析表明,差异表达蛋白参与了细胞质翻译、蛋白质稳定、干细胞群体维持、细胞迁移等多个生物学过程相关通路;与对照组相比,成球实验和迁移实验表明敲降S6K1可显著抑制乳腺癌细胞干性表型和迁移能力(P<0.01).结论:S6K1在乳腺癌细胞中调控多个生物学过程相关蛋白的表达,敲降S6K1明显抑制了乳腺癌细胞的干性和迁移能力,表明S6K1可能成为乳腺癌治疗的新靶点.

OBJECTIVE:To investigate the expression and regulation of the ribosomal protein S6 kinase β-1(S6K1)on the malignant phenotype of breast cancer cells.METHODS:Amplification and expression of S6K1 in breast cancer tissues were analyzed by cBioPortal and GEPIA databases,and their relationships with clinical characteristics of the breast cancers were also analyzed.Small interfering RNA was employed to knock down S6K1 in the breast cancer MCF7 cells in vitro to investigate its impact on protein expression profile using mass spectrometry,stemness of breast cancer cells using the sphere formation assay,and migration ability using the migration assay.RESULTS:S6K1 gene exhibited frequent amplification in breast cancer tissues,with a higher amplification rate in invasive compared with non-invasive breast cancers(P<0.05),and with significant relationship to poor prognosis of patients(P<0.01).In addition,the expression was higher in breast cancer tissues compared with adjacent tissues(P<0.01),while the expression levels did not differ significantly among different stages of breast cancer(P>0.05).Mass spectrometry revealed 251 upregulated and 224 downregulated proteins following the S6K1 knockdown in the MCF7 cells.GO pathway enrichment analysis indicated that S6K1 regulated the expression of multiple proteins involved in biological processes such as cytoplasmic translation,protein stabilization,stem cell population maintenance,and cell migration.Additional assays demonstrated that S6K1 knockdown significantly inhibited the stemness phenotype and migration ability of breast cancer cells(P<0.05).CONCLUSION:S6K1 regulated the expression of proteins involved in multiple biological processes in breast cancer cells in vivo,and the knockdown of S6K1 significantly inhibited the stemness and migration ability of breast cancer cells in vitro,indicating a crucial regulatory role for S6K1 and highlighting its potential as a novel therapeutic target for breast cancer therapy.

舒杏梅;石小倩;刘燕;李丹

国家癌症中心/国家肿瘤临床医学研究中心/中国医学科学院北京协和医学院肿瘤医院,分子肿瘤学国家重点实验室,北京 100021

临床医学

核糖体蛋白S6激酶β-1乳腺癌细胞干性迁移蛋白组学

S6K1breast cancerstemness of cellsmigrationproteomics

《癌变·畸变·突变》 2024 (003)

中心体lncRNA CAT1在食管癌细胞周期调控中的机制研究

179-186,223 / 9

国家自然科学基金(81972318)

10.3969/j.issn.1004-616x.2024.03.003

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