KDM5B在宫颈癌中的表达意义及对宫颈癌干细胞的影响  

Expression of KDM5B in cervical cancer and its effect on cervical cancer stem cells

作　　者：杨宝娟[1] 张庆[1] 王志红[1] 王武亮[1] 张珂[1] YANG Bao-Juan;ZHANG Qing;WANG Zhi-Hong;WANG Wu-Liang;ZHANG Ke(Department of Obstetrics and Gynecology,Second Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,China)

机构地区：[1]郑州大学第二附属医院妇产科,郑州450000

出　　处：《中国免疫学杂志》2020年第23期2879-2885,共7页Chinese Journal of Immunology

摘　　要：目的:研究组蛋白去甲基化转移酶(KDM5B)在宫颈癌组织中的表达,及其与患者临床病理特征和预后之间的关系,并探讨KDM5B对宫颈癌干细胞(CCSCs)的影响。方法:免疫组化检测KDM5B在70例宫颈癌组织和30例正常宫颈组织中的表达水平,并分析宫颈癌组织中KDM5B的表达与临床病理特征的关系及对患者预后的影响。流式细胞术分选Caski细胞系的肿瘤干细胞。Western blot检测KDM5B在CCSCs中的表达;经脂质体分别转染KDM5B siRNA(si-KDM5B)和对照(si-NC)48 h后,MTS检测各组CCSCs增殖能力,软琼脂克隆形成实验检测各组CCSCs肿瘤球形成能力,Transwell实验检测各组CCSCs转移能力,Western blot检测各组CCSCs中OCT4、Nanog、SOX2干性基因蛋白的表达;KDM5B干扰质粒及对照NC感染CCSCs,经1.0μg/ml遗传霉素G418筛选稳定敲减KDM5B的CCSCs细胞,注射到BALB/c裸鼠皮下,观察KDM5B对CCSCs体内致瘤性的影响。结果:免疫组化结果显示KDM5B在宫颈癌组织中的表达显著高于正常宫颈组织,其阳性表达与宫颈癌肿瘤大小及淋巴结转移显著相关(P<0.05),宫颈癌组织中KDM5B阳性表达患者生存期较短。KDM5B在CCSCs中的表达显著高于宫颈癌细胞Caski和人宫颈上皮永生化细胞H8(P<0.05);转染KDM5B siRNA后,CCSCs细胞增殖、软琼脂克隆形成及转移能力均下降,CCSCssh-KDM5B裸鼠成瘤体积显著小于CCSCsNC组(P<0.05)。结论:KDM5B在宫颈癌组织和CSCs中均高表达,且阳性表达与肿瘤大小、淋巴结转移及预后不良相关,同时干扰KDM5B的表达抑制CCSCs的干性,KDM5B可以作为治疗宫颈癌的潜在分子靶点。Objective:To detect the relationship between the expression of lysine demethylation 5B(KDM5B)in cervical cancer tissues and the clinical features and prognosis of cervical cancer patients,and to explore the effect of KDM5B on cervical cancer stem cells(CCSCs).Methods:The expression of KDM5B in 70 cases of cervical cancer tissues and 30 cases of normal cervical tissues was detected by immunohistochemistry.The relationship between KDM5B expression and clinicopathological features and the prognosis of cervical cancer tissues were analyzed.Flow cytometry was used to sort cancer stem cells from the Caski cell line.The expression of KDM5B in CCSCs was detected by Western blot.After liposome transfected with KDM5B siRNA(si-KDM5B)and control(si-NC)for 48 h,the proliferation of CCSCs in each group was detected by MTS.Tumor sphere formation ability of CCSCs in each group was detected by soft agar colony formation assay.The metastasis ability of CCSCs in each group was detected by Transwell test.The expression of OCC4,Nanog and SOX2 stem gene proteins in CCSCs was detected by Western blot.The KDM5B interference plasmid and control NC infected CCSCs.CCSCs cells stably knocked down KDM5B were screened by 1.0μg/ml geneticin G418,the cells were injected subcutaneously into nude mice to observed the effect of KDM5B on the tumorigenicity of CCSCs in vivo.Results:The results of immunohistochemistry showed that the expression of KDM5B in cervical cancer tissues was significantly higher than that in normal cervical tissues.The positive expression was significantly correlated with tumor size and lymph node metastasis of cervical cancer(P<0.05),and patients with positive expression of KDM5B had a shorter survival period.The expression of KDM5B in CCSCs was significantly higher than that of cervical cancer cells Caski and human cervical epithelial cells H8(P<0.05).After transfection of KDM5B siRNA,CCSCs cell proliferation,soft agar colony formation and metastasis ability were decreased.The tumor formation volume of CCSCssh-KDM5B nude

关 键 词：宫颈癌 KDM5B 肿瘤干细胞 干性 

分 类 号：R737.33[医药卫生—肿瘤]