Ander Martin, Daniela Gerovska, Marcos J Arauzo-Bravo, Maitane Duarte García-Escudero, Helena García García, Iratxe Bañares, Naroa Fontal, Geraldine Siegfried, Serge Evrad, Simon Pernot, Abdel-Majid Khatib, Iker Badiola
{"title":"抑制PCSK9可减弱结直肠癌干细胞在肝转移过程中诱导的肝内皮细胞活化","authors":"Ander Martin, Daniela Gerovska, Marcos J Arauzo-Bravo, Maitane Duarte García-Escudero, Helena García García, Iratxe Bañares, Naroa Fontal, Geraldine Siegfried, Serge Evrad, Simon Pernot, Abdel-Majid Khatib, Iker Badiola","doi":"10.3390/cancers17121977","DOIUrl":null,"url":null,"abstract":"<p><p><b>Background:</b> Colorectal cancer (CRC) is among the most prevalent and lethal cancers globally, with liver metastasis representing the leading cause of CRC-related mortality. Proprotein convertase subtilisin/kexin type 9 (PCSK9) has recently gained attention due to its overexpression in colorectal tumor tissues and its potential role in driving metastatic progression. This aims to investigate the involvement of PCSK9 in the liver metastatic niche, focusing on its effects on liver sinusoidal endothelial cells (LSECs), key components of the liver microenvironment. <b>Methods:</b> LSECs were stimulated with conditioned media derived from differentiated colorectal cancer cells and cancer stem cells (CSCs), the latter generated by reprogramming SW620 and CT26 cell lines. RNA sequencing was used to profile gene expression in LSECs. PCSK9 mRNA and protein levels were quantified by qPCR and Western blotting, respectively. PCSK9 expression in CRC liver metastases was evaluated by immunofluorescent staining. <b>Results</b>: PCSK9 was detected in both human and murine LSECs and significantly upregulated following exposure to CSC-conditioned media. Immunofluorescent staining confirmed PCSK9 expression in LSECs within CRC liver metastases. Total RNA sequencing revealed that a pre-treatment of LSECs with the PCSK9 inhibitor PF-06446864 prior to CSC stimulation seems to reduce the expression of microRNAs linked to cell migration and proliferation. Functional assays demonstrated that CSC-conditioned media enhanced LSEC proliferation and migration, effects reversed by PCSK9 inhibition. <b>Conclusions:</b> PCSK9 promotes the activation of LSECs in response to colorectal CSCs, contributing to a pro-metastatic phenotype. These findings highlight PCSK9 as a potential therapeutic target in colorectal liver metastasis.</p>","PeriodicalId":9681,"journal":{"name":"Cancers","volume":"17 12","pages":""},"PeriodicalIF":4.4000,"publicationDate":"2025-06-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12190733/pdf/","citationCount":"0","resultStr":"{\"title\":\"Inhibition of PCSK9 Attenuates Liver Endothelial Cell Activation Induced by Colorectal Cancer Stem Cells During Liver Metastasis.\",\"authors\":\"Ander Martin, Daniela Gerovska, Marcos J Arauzo-Bravo, Maitane Duarte García-Escudero, Helena García García, Iratxe Bañares, Naroa Fontal, Geraldine Siegfried, Serge Evrad, Simon Pernot, Abdel-Majid Khatib, Iker Badiola\",\"doi\":\"10.3390/cancers17121977\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p><b>Background:</b> Colorectal cancer (CRC) is among the most prevalent and lethal cancers globally, with liver metastasis representing the leading cause of CRC-related mortality. Proprotein convertase subtilisin/kexin type 9 (PCSK9) has recently gained attention due to its overexpression in colorectal tumor tissues and its potential role in driving metastatic progression. This aims to investigate the involvement of PCSK9 in the liver metastatic niche, focusing on its effects on liver sinusoidal endothelial cells (LSECs), key components of the liver microenvironment. <b>Methods:</b> LSECs were stimulated with conditioned media derived from differentiated colorectal cancer cells and cancer stem cells (CSCs), the latter generated by reprogramming SW620 and CT26 cell lines. RNA sequencing was used to profile gene expression in LSECs. PCSK9 mRNA and protein levels were quantified by qPCR and Western blotting, respectively. PCSK9 expression in CRC liver metastases was evaluated by immunofluorescent staining. <b>Results</b>: PCSK9 was detected in both human and murine LSECs and significantly upregulated following exposure to CSC-conditioned media. Immunofluorescent staining confirmed PCSK9 expression in LSECs within CRC liver metastases. Total RNA sequencing revealed that a pre-treatment of LSECs with the PCSK9 inhibitor PF-06446864 prior to CSC stimulation seems to reduce the expression of microRNAs linked to cell migration and proliferation. Functional assays demonstrated that CSC-conditioned media enhanced LSEC proliferation and migration, effects reversed by PCSK9 inhibition. <b>Conclusions:</b> PCSK9 promotes the activation of LSECs in response to colorectal CSCs, contributing to a pro-metastatic phenotype. These findings highlight PCSK9 as a potential therapeutic target in colorectal liver metastasis.</p>\",\"PeriodicalId\":9681,\"journal\":{\"name\":\"Cancers\",\"volume\":\"17 12\",\"pages\":\"\"},\"PeriodicalIF\":4.4000,\"publicationDate\":\"2025-06-13\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://www.ncbi.nlm.nih.gov/pmc/articles/PMC12190733/pdf/\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Cancers\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.3390/cancers17121977\",\"RegionNum\":2,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q1\",\"JCRName\":\"ONCOLOGY\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Cancers","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.3390/cancers17121977","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q1","JCRName":"ONCOLOGY","Score":null,"Total":0}
引用次数: 0
摘要
背景:结直肠癌(CRC)是全球最常见和最致命的癌症之一,肝转移是CRC相关死亡的主要原因。Proprotein convertase subtilisin/kexin type 9 (PCSK9)最近因其在结直肠肿瘤组织中的过度表达及其在驱动转移进展中的潜在作用而受到关注。本研究旨在研究PCSK9在肝转移生态位中的作用,重点关注其对肝窦内皮细胞(LSECs)的影响,肝窦内皮细胞是肝微环境的关键组成部分。方法:用分化的结直肠癌细胞和癌症干细胞(CSCs)的条件培养基刺激LSECs,后者是通过重编程SW620和CT26细胞系产生的。采用RNA测序分析LSECs的基因表达。采用qPCR和Western blotting分别检测PCSK9 mRNA和蛋白水平。免疫荧光染色法检测PCSK9在结直肠癌肝转移灶中的表达。结果:PCSK9在人和小鼠LSECs中均检测到,并在暴露于csc条件培养基后显著上调。免疫荧光染色证实PCSK9在结直肠癌肝转移的LSECs中表达。总RNA测序显示,在CSC刺激之前用PCSK9抑制剂PF-06446864预处理LSECs似乎可以降低与细胞迁移和增殖相关的microrna的表达。功能分析表明csc条件培养基增强了LSEC的增殖和迁移,这种作用被PCSK9抑制逆转。结论:PCSK9促进LSECs对结直肠CSCs的激活,促成了一种促转移表型。这些发现突出了PCSK9作为结直肠癌肝转移的潜在治疗靶点。
Inhibition of PCSK9 Attenuates Liver Endothelial Cell Activation Induced by Colorectal Cancer Stem Cells During Liver Metastasis.
Background: Colorectal cancer (CRC) is among the most prevalent and lethal cancers globally, with liver metastasis representing the leading cause of CRC-related mortality. Proprotein convertase subtilisin/kexin type 9 (PCSK9) has recently gained attention due to its overexpression in colorectal tumor tissues and its potential role in driving metastatic progression. This aims to investigate the involvement of PCSK9 in the liver metastatic niche, focusing on its effects on liver sinusoidal endothelial cells (LSECs), key components of the liver microenvironment. Methods: LSECs were stimulated with conditioned media derived from differentiated colorectal cancer cells and cancer stem cells (CSCs), the latter generated by reprogramming SW620 and CT26 cell lines. RNA sequencing was used to profile gene expression in LSECs. PCSK9 mRNA and protein levels were quantified by qPCR and Western blotting, respectively. PCSK9 expression in CRC liver metastases was evaluated by immunofluorescent staining. Results: PCSK9 was detected in both human and murine LSECs and significantly upregulated following exposure to CSC-conditioned media. Immunofluorescent staining confirmed PCSK9 expression in LSECs within CRC liver metastases. Total RNA sequencing revealed that a pre-treatment of LSECs with the PCSK9 inhibitor PF-06446864 prior to CSC stimulation seems to reduce the expression of microRNAs linked to cell migration and proliferation. Functional assays demonstrated that CSC-conditioned media enhanced LSEC proliferation and migration, effects reversed by PCSK9 inhibition. Conclusions: PCSK9 promotes the activation of LSECs in response to colorectal CSCs, contributing to a pro-metastatic phenotype. These findings highlight PCSK9 as a potential therapeutic target in colorectal liver metastasis.
期刊介绍:
Cancers (ISSN 2072-6694) is an international, peer-reviewed open access journal on oncology. It publishes reviews, regular research papers and short communications. Our aim is to encourage scientists to publish their experimental and theoretical results in as much detail as possible. There is no restriction on the length of the papers. The full experimental details must be provided so that the results can be reproduced.