敲低Hsa_circ_0020080通过调节脂代谢对人乳腺癌细胞增殖、迁移及侵袭的促进作用  

Knockdown of Hsa_Circ_0020080 Promotes Proliferation,Migration and Invasion of Human Breast Cancer Cells by Regulating Lipid Metabolism

作　　者：王凌霞 李晓 徐诗靓 王波[1] 杨欢[1] WANG Lingxia;LI Xiao;XU Shiliang;WANG Bo;YANG Huan(The Second Affiliated Hospital of Soochow University,Suzhou,Jiangsu 215000)

机构地区：[1]苏州大学附属第二医院,江苏苏州215000

出　　处：《临床输血与检验》2025年第1期34-48,共15页Journal of Clinical Transfusion and Laboratory Medicine

基　　金：国家自然科学基金青年项目(No.81702078);姑苏卫生人才计划人才科研项目(No.GSWS2023041);苏州市卫生青年骨干人才“全国导师制”项目(No.Qngg2023008);苏州市科技计划项目(No.SKY2021045、No.SKJY2021097);苏州大学附属第二医院预研项目(No.SDFEYBS2218)资助。

摘　　要：目的本研究旨在探讨敲低Hsa_circ_0020080通过调节脂代谢对人乳腺癌细胞增殖、迁移和侵袭行为的影响。方法定量实时荧光PCR(qRT-PCR)验证Hsa_circ_0020080敲低效率及MDM2、VEGF、FASN、HMGCR、CPT1A、SREBP1基因表达水平。Western blot检测MDM2、VEGF蛋白表达水平。体外采用克隆形成和CCK8实验检测细胞增殖功能;Transwell迁移和基质胶侵袭实验检测细胞迁移和侵袭功能。体内构建裸鼠乳房脂肪垫下移植瘤。免疫组化检测MDM2、VEGF、FASN、HMGCR、SREBP1蛋白表达。油红O染色和甘油三酯试剂盒检测甘油三酯表达量。利用脂肪酸合成酶FASN有效抑制剂(C75)进行增殖、迁移和侵袭相关的功能回补实验。结果体外敲低Hsa_circ_0020080能够增加乳腺癌细胞克隆形成、增殖、迁移及侵袭能力,促进MDM2、VEGF、FASN、HMGCR、CPT1A、SREBP1基因及MDM2、VEGF蛋白表达,增加乳腺癌细胞甘油三酯含量;体内敲低Hsa_circ_0020080促进裸鼠移植瘤的生长,增加MDM2、VEGF、FASN、HMGCR、SREBP1蛋白表达及裸鼠移植瘤组织甘油三酯水平。C75能部分逆转敲低Hsa_circ_0020080对乳腺癌细胞增殖、迁移及侵袭功能的促进作用。结论敲低Hsa_circ_0020080可以通过脂代谢促进乳腺癌细胞的增殖、侵袭及迁移功能,提示Hsa_circ_0020080在乳腺癌发展中发挥着重要的生物学作用。Objective The purpose of this study was to explore the effect of knockdown of Hsa_circ_0020080 on the proliferation,migration and invasion of human breast cancer cells by regulating lipid metabolism.Methods Quantitative real-time fluorescence PCR(qRT PCR)was used to validate the efficiency of Hsa_circ_0020080 knockdown and the expression levels of MDM2,VEGF,FASN,HMGCR,CPT1A,SREBP1.Western blot was used to detect the expression levels of MDM2 and VEGF proteins.Cell proliferation function was examined in vitro using colony formation and CCK8 assays;Transwell migration and matrigel invasion experiments were conducted to detect cell migration and invasion functions.Transplanted tumor model of nude mice was established in vivo.Immunohistochemistry was used to detect the protein expression of MDM2,VEGF,FASN,HMGCR,and SREBP1ere detected by.Oil red O staining and triglyceride assay kit were used to detect the expression levels of triglyceride.Functional rescue experiments were conducted by the effective inhibitor of fatty acid synthase FASN(C75).Results The Knockdown of Hsa_circ_0020080 in vitro could increase the colony formation,proliferation,migration and invasion ability of breast cancer cells,promote the expression of MDM2,VEGF,FASN,HMGCR,CPT1A,SREBP1 genes and MDM2,VEGF proteins,and increase the triglyceride content of breast cancer cells;The Knockdown of Hsa_circ_0020080 in vivo promotes the growth of xenograft tumors in nude mice,increases the expression of MDM2,VEGF,FASN,HMGCR,SREBP1 proteins,and triglyceride levels in xenograft tumors of nude mice.C75 can partially rescue proliferation,migration and invasion ability of breast cancer cells with the knockdown of Hsa_circ_0020080.Conclusion Knockdown of Hsa_circ_0020080 promotes the proliferation,invasion and migration of breast cancer cells through lipid metabolism,which plays an important biological role in the development of breast cancer.

关 键 词：Hsa_(c)irc_(0)020080 乳腺癌 增殖 迁移 侵袭 脂代谢 

分 类 号：R737.9[医药卫生—肿瘤]