肿瘤相关巨噬细胞极化状态对前列腺癌干细胞自我更新能力和血管生成拟态的影响  

Effects of polarization state of tumor-associated macrophages on self-renewal ability and vasculogenic mimicry of prostate cancer stem cells

作　　者：田野[1] 阿不都米吉提·阿不都克力木 王鹏[1] 沙漠[1] 崔崎[1] TIAN Ye;ABUDU MIJITI·Abudu Kelimu;WANG Peng;SHA Mo;CUI Qi(Department of Urinary Surgery,Fourth Affiliated Hospital,Xinjiang Medical University,Xinjiang Uygur Autonomous Region Hospital of Traditional Chinese Medicine,Urumqi 830000,China)

机构地区：[1]新疆医科大学第四附属医院新疆维吾尔自治区中医院泌尿外科,新疆乌鲁木齐830000

出　　处：《吉林大学学报（医学版）》2022年第2期374-382,共9页Journal of Jilin University：Medicine Edition

基　　金：新疆维吾尔自治区科技厅自然科学基金项目(2019D01C114)。

摘　　要：目的:探讨肿瘤相关巨噬细胞(TAM)极化状态对前列腺癌(PCa)干细胞自我更新能力与血管生成拟态的影响,阐明TAM在PCa进展中的作用。方法:从人PCa细胞系DU145中分选出PCa干细胞,将人单核白血病细胞株THP-1诱导为M2型TAM,作为诱导组,并以未诱导的THP-1细胞作为对照组,实时荧光定量PCR(RT-qPCR)法检测细胞中诱导型一氧化氮合酶(iNOS)和重组人精氨酸酶1(Arg-1)mRNA表达水平。实验分为PCa-肿瘤干细胞(CSC)组(常规培养基培养PCa干细胞)、THP-1+PCa-CSC组(未诱导的THP-1细胞的培养液上清培养PCa干细胞)和TAM+PCa-CSC组(诱导处理的THP-1细胞的TAM条件培养基培养PCa干细胞),模拟微环境建立TAM与PCa干细胞共培养体系,培养48 h后,通过细胞成球实验检测各组PCa干细胞成球情况,细胞克隆形成实验检测各组PCa干细胞集落数目,细胞划痕实验检测各组PCa干细胞划痕愈合率,Transwell小室实验检测各组PCa干细胞侵袭数目,血管生成拟态形成实验检测各组PCa干细胞管状结构数目。结果:人PCa细胞系DU145经分选后获得CD44+CD133+标记的干细胞。与对照组比较,诱导组THP-1细胞形态呈不规则多边形,iNOS mRNA表达水平降低(t=21.021,P<0.05),Arg-1 mRNA表达水平升高(t=26.153,P<0.05),CD86蛋白表达水平降低(t=34.556,P<0.05),CD206蛋白表达水平升高(t=31.095,P<0.05);与PCa-CSC组比较,THP-1+PCa-CSC组和TAM+PCa-CSC组细胞球体积变大,细胞集落数目增加(P<0.05),划痕愈合率升高(P<0.05),细胞侵袭数目增加(P<0.05),细胞管状结构数目增加(P<0.05);与THP-1+PCa-CSC组比较,TAM+PCa-CSC组细胞球体积进一步变大,细胞集落数目和细胞侵袭数目均增加(P<0.05),划痕愈合率升高(P<0.05),细胞管状结构数目增加(P<0.05)。结论:通过调控TAM极化状态可诱导PCa干细胞转移,进一步促进其自我更新与血管生成拟态形成。Objective:To explore the effects of tumor-associated macrophages(TAM)polarization state on the self-renewal ability and vasculogenic mimicry of prostate cancer(Pca)stem cells,and to clarify the effect of TAM in the development of PCa.Methods:The PCa stem cells were selected from human PCa cell line DU145,and the human mononuclear leukemia cell strain THP-1 was induced into the M2 TAM,which was used as induction group,and the uninduced THP-1 cells were used as control group;real-time fluorescence quantitative PCR(RT-qPCR)method was used to detect the expression levels of inducible nitric oxide synthase(iNOS)mRNA and recombinant human arginase-1(Arg-1)mRNA in the cells.The experiment was divided into PCa-cancer stem cells(CSC)group(PCa stem cells were cultured in the conventional culture medium),THP-1+PCa-CSC group(PCa stem cells were cultured in the culture supernatant of uninduced THP-1 cells)and TAM+PCa-CSC group(PCa stem cells were cultured in TAM-conditioned medium of induced THP-1 cells);a co-culture system of TAM and PCa stem cells was established by simulating the microenvironment.After 48 h of culture,the spheroidization of PCa stem cells in various groups was detected by cell spheroidization experiment,the number of cell colonies of PCa stem cells in various groups was detected by cell clone formation experiment,cell scratch test was used detect the scratch healing rates of cells in various groups,and Transwell chamber test was used to detect the number of invasion PCa stem cells in various groups;vasculogenic mimicry formation experiment was used to detect the number of tubular structures of PCa stem cells in various groups.Results:The human PCa DU145 cells were sorted to obtain the CD44+CD133+labeled prostate cancer stem cells.Compared with control group,the THP-1 cells in induction group showed irregular polygonal shapes,the expression level of iNOS mRNA was decreased(t=21.021,P<0.05),the expression level of Arg-1 mRNA was increased(t=26.153,P<0.05),the expression level of CD86 protein was decreased(t=

关 键 词：肿瘤相关巨噬细胞 前列腺癌干细胞 自我更新 血管生成拟态 

分 类 号：R737.25[医药卫生—肿瘤]