细胞外基质硬度对人前列腺癌细胞可塑性调控作用的研究  

Regulatory effect of extracellular matrix stiffness on the plasticity of human prostate cancer cells

作　　者：郑雨 马磊[2] 李睿智[2] 牟洁 李菁 王栋[2] 祝海[3] ZHENG Yu;MA Lei;LI Ruizhi;MU Jie;LI Jing;WANG Dong;ZHU Hai(Faculty of Medicine,Qingdao University,Qingdao 266071,China)

机构地区：[1]青岛大学医学部,山东青岛266071 [2]青岛大学转化医学研究院 [3]青岛市市立医院泌尿外科 [4]青岛大学药学院

出　　处：《精准医学杂志》2024年第3期222-225,231,共5页Journal of Precision Medicine

基　　金：山东省自然科学基金资助项目(ZR2023MH327);青岛市自然科学基金资助项目(23-2-1-193-zyyd-jch)。

摘　　要：目的探讨细胞外基质硬度对人前列腺癌细胞LNCaP可塑性调控的作用。方法将人LNCaP细胞分别于杨氏模量为3 GPa(A组)、20 kPa(B组)、6 kPa(C组)和1 kPa(D组)四种不同硬度细胞外基质培养皿中培养1周。采用明场显微镜及荧光显微镜观察各组细胞在不同硬度基底上的形态;采用实时荧光定量聚合酶链反应(RT-qPCR)技术检测各组细胞的管腔细胞标志物基因AR、CK 8、CK 18、PSA,基底细胞标志物基因CK 5、P 63,以及增殖基因KI 67、PCNA的表达水平。结果明场显微镜及荧光显微镜观察结果显示,A组人LNCaP细胞呈现典型铺展上皮细胞形态,而B~D组均呈现团聚小细胞片状态,且D组细胞形成了三维类器官结构。RT-qPCR结果显示,四组人LNCaP细胞中KI 67、P 63基因表达水平无显著差异(P>0.05);B组PCNA基因表达水平显著高于A、C组(F=34.96,t=8.39、6.37,P<0.05);B组CK 5基因表达水平显著高于其他三组(F=29.35,t=4.46~6.73,P<0.05);D组AR、CK 8、CK 18、PSA基因表达水平显著高于其他三组(F=13.66~56.43,t=3.03~11.51,P<0.05)。结论硬度较低(杨氏模量1 kPa)的细胞外基质环境能较好维持人LNCaP细胞的管腔细胞特征,而硬度较高(杨氏模量20 kPa)的细胞外基质能使人LNCaP细胞呈现出中间态细胞表型,或许是导致前列腺癌发生的因素之一。Objective To investigate the regulatory effect of extracellular matrix(ECM)stiffness on the plasticity of human prostate cancer cells(LNCaP).Methods Human LNCaP cells were cultured in ECM culture dishes with four types of stiffness(i.e.,Young’s modulus of 3 GPa[group A],20 kPa[group B],6 kPa[group C],and 1 kPa[group D])for one week.The morphology of cells on substrates of different stiffness values was observed for each group using bright-field microscopy and fluorescence microscopy.Reverse transcription-quantitative polymerase chain reaction(RT-qPCR)was used to measure the expression levels of luminal cell marker genes(AR,CK 8,CK 18,and PSA),basal cell marker genes(CK 5 and P 63),and proliferative genes(KI 67 and PCNA)in each group.Results Bright-field microscopy and fluorescence microscopy showed that human LNCaP cells in group A exhibited typically spread epithelial cell morphology,whereas the cells in groups B to D showed the state of agglomerated small cell sheets,and the cells in group D formed a three-dimensional organ-like structure.RT-qPCR results showed that there were no significant differences between the four groups in the expression levels of KI 67 and P 63 genes in human LNCaP cells(P>0.05).The expression level of PCNA gene in group B was significantly higher than that in groups A and C(F=34.96,t=8.39,6.37,P<0.05).The expression level of CK 5 gene in group B was significantly higher than that in the other three groups(F=29.35,t=4.46-6.73,P<0.05).The expression levels of AR,CK 8,CK 18,and PSA genes in group D were significantly higher than those in the other three groups(F=13.66-56.43,t=3.03-11.51,P<0.05).Conclusion ECM environment with a lower stiffness value(Young’s modulus of 1 kPa)can better maintain the luminal cell characteristics of human LNCaP cells,whereas ECM with a higher stiffness value(Young’s modulus of 20 kPa)causes human LNCaP cells to exhibit an intermediate state cellular phenotype,which may be one of the factors leading to the development of prostate cancer.

关 键 词：前列腺肿瘤 细胞系 肿瘤 细胞外基质 细胞可塑性 

分 类 号：R737.25[医药卫生—肿瘤] R329.24[医药卫生—临床医学]