HIF-1a/PDK1通路活化促进化疗后残存乳腺癌组织癌干细胞生成  

Activation of HIF-1a/PDK1 Pathway Promotes the Generation of Cancer Stem Cells in Residual Breast Cancer Tissue after Chemotherapy

作　　者：朱芳 王佑权 徐喆 王先明[3] 屈洪波 Zhu Fang;Wang Youquan;Xu Zhe;Wang Xianming;Qu Hongbo(Department of Health Management,the People’s Hospital of Longhua,Shenzhen,Shenzhen 518000,Guangdong,China;Department of Thyroid and Breast Surgery,the People’s Hospital of Longhua,Shenzhen,Shenzhen 518000,Guangdong,China;Department of Thyroid and Breast Surgery,South China Hospital Affiliated to Shenzhen University,Shenzhen 518000,Guangdong,China)

机构地区：[1]深圳市龙华区人民医院健康管理科,广东深圳518000 [2]深圳市龙华区人民医院甲乳外科,广东深圳518000 [3]深圳大学附属华南医院甲乳外科,广东深圳518000

出　　处：《肿瘤预防与治疗》2023年第10期819-825,共7页Journal of Cancer Control And Treatment

基　　金：湖南省科技厅临床医疗技术创新引导项目(编号:2020SK50308)。

摘　　要：目的:探讨新辅助化疗后残存乳腺癌组织低氧诱导因子(hypoxia inducible factor 1a,HIF-1a)/丙酮酸脱氢酶激酶同工酶1(pyruvate dehydrogenase kinase isoenzyme 1,PDK1)信号通路活化是否促进乳腺癌干细胞(breast cancer stem cells,BCSCs)的生成。方法:实验分为化疗前乳腺癌组织组和化疗后残存乳腺癌组织组。免疫组织化学染色检测两组细胞中HIF-1a、PDK1及BCSCs标志物乙醛脱氢酶1A1(acetaldehyde dehydrogenase 1A1,ALDH1A1)表达。通过无血清悬浮培养获得BCSCs微球体和观察微球体形态学;有限稀释法实验和Transwell体外侵袭实验分别检测微球体细胞单克隆形成能力和侵袭潜能;Western blot检测微球体细胞HIF-1a、PDK1及ALDH1A1蛋白表达水平。结果:免疫组化结果显示化疗后残存乳腺癌组织较化疗前乳腺癌组织高表达HIF-1a、PDK1及ALDH1A1蛋白,差异有统计学意义(P<0.05),ALDH1A1与HIF-1a和PDK1的表达相关。无血清悬浮培养获得BCSCs微球体为后续实验奠定基础,有限稀释法检测显示化疗后残存乳腺癌组织中BCSCs含量是化疗前2.5倍,同时微球体细胞侵袭能力明显增强。Western blot结果显示BCSCs微球体细胞高表达HIF-1a、PDK1及ALDH1A1蛋白,与化疗前乳腺癌组织相比,表达差异有统计学意义(P<0.05)。结论:新辅助化疗促进残存乳腺癌组织中BCSCs微球体细胞富集,其可能通过激活HIF-1a/PDK1信号通路并促进ALDH1A1表达,从而获得癌干细胞潜能。Objective:To investigate whether the activation of hypoxia inducible factor 1a(HIF-1a)/pyruvate dehydrogenase kinase isoenzyme 1(PDK1)signaling pathway in residual breast cancer tissue after neoadjuvant chemotherapy promotes breast cancer stem cells(BCSCs)generation.Methods:There were two groups,the pre-chemotherapy breast cancer tissue group and the post-chemotherapy residual breast cancer tissue group,in the experiment.The expressions of HIF-1a,PDK1 and BCSCs marker,acetaldehyde dehydrogenase 1A1(ALDH1A1),were detected by immunohistochemical staining.BCSCs microspheres were obtained by serum-free suspension culture and their morphology was observed.Limited dilution assay and Transwell assay were used to detect the clone-forming ability and invasion potential of microspheres.The expression levels of HIF-1a,PDK1 and ALDH1A1 proteins in microspheres were detected by Western blot.Results:Immunohistochemical results showed that the expressions of HIF-1a,PDK1 and ALDH1A1 proteins in breast cancer tissue after chemotherapy were significantly higher than those before chemotherapy(P 0.05),and the expression of ALDH1A1 was correlated to the expressions of HIF-1a and PDK1.The BCSCs microspheres obtained by serum-free suspension culture laid the foundation for subsequent experiments.The limited dilution assay showed that the BCSCs content in breast cancer tissue after chemotherapy was 2.5 times as much as that before chemotherapy,and the invasion ability of microspheres was significantly enhanced after chemotherapy.Western blot results showed that HIF-1a,PDK1 and ALDH1A1 proteins were highly expressed in BCSCs microspheres,and the differences were statistically significant in the expression levels before and after chemotherapy(P 0.05).Conclusion:Chemotherapy promotes the enrichment of BCSCs microspheres in residual breast cancer tissue,which may obtain the potential of cancer stem cells by activating HIF-1a/PDK1 signaling pathway and promoting the expression of ALDH1A1.

关 键 词：乳腺癌干细胞 低氧微环境 HIF-1A PDK1 ALDH1A1 

分 类 号：Q74[生物学—分子生物学] R737.9[医药卫生—肿瘤]