基于细胞外囊泡DNA甲基化标志物探讨子宫颈癌侵袭转移的风险  

Exploring the risk of tumor invasion and metastasis in cervical cancer based on extracellular vesicle DNA methylation biomarkers

作　　者：何薇薇[1] 赵妍 李能[1] 谢琦 吴菲[1] He Weiwei;Zhao Yan;Li Neng;Xie Qi;Wu Fei(Department of Gynecology,Hunan Provincial Maternal and Child Health Care Hospital/Hunan Institute of Reproductive Medicine,Changsha 410000,China)

机构地区：[1]湖南省妇幼保健院/湖南省生殖医学研究院妇科,长沙410000

出　　处：《临床与实验病理学杂志》2025年第4期474-482,共9页Chinese Journal of Clinical and Experimental Pathology

基　　金：湖南省卫生健康委员会科研项目(202202115219)。

摘　　要：目的探讨子宫颈癌癌细胞外囊泡(extracellular vesicles,EVs)中甲基化调节剂核内不均一核糖核蛋白C(heterogeneous nuclear ribonucleoprotein C,HNRNPC)对肿瘤侵袭转移的预测价值。方法收集人正常子宫颈上皮细胞系(HcerEpic)、3种人子宫颈癌细胞系(HeLa、SiHa和CaSki)和人脐静脉血管内皮细胞(human umbilical vein endothelial cells,HUVECs)用于体外实验。采用Western blot法分析细胞中HNRNPC蛋白表达。应用sh-NC和sh-HNRNPC转染CaSki细胞,并从细胞培养上清液中分离EVs,分别考察EVs对SiHa细胞增殖、迁移和侵袭的影响,和对HUVECs血管生成的影响。对于组织微阵列检测,从本院获得正常子宫颈组织(n=8)、低级别(n=32)和高级别(n=37)鳞状上皮内病变和子宫颈癌(n=153)标本。根据HNRNPC免疫评分将子宫颈癌患者分为HNRNPC高表达组(n=99)和低表达组(n=54),比较两组的临床病理特征和预后。结果与HcerEpic细胞相比,SiHa、HeLa、CaSki细胞中HNRNPC表达增加。在用CaSki-EVs处理的SiHa细胞中,与sh-NC-EVs组相比,sh-HNRNPC-EVs组SiHa细胞的增殖能力、细胞集落数以及迁移和侵袭细胞数均显著降低(P<0.05)。在用CaSki-EVs处理的HUVECs细胞中,与sh-NC-EVs组相比,sh-HNRNPC-EVs组HUVECs细胞的PCNA、VEGFA蛋白表达、血管生成长度均显著降低(P<0.05)。HNRNPC表达随着子宫颈上皮细胞的转化而逐渐增加(F=106.9,P<0.001),且与正常组织相比,高级别鳞状上皮内病变和子宫颈癌中HNRNPC表达显著增加(P<0.001)。HNRNPC与子宫颈癌细胞分化程度、肿瘤大小、宫旁浸润、阴道浸润、晚期FIGO分期和盆腔淋巴结转移相关(P<0.05)。Kaplan-Meier分析显示,与HNRNPC低表达组相比,HNRNPC高表达组子宫颈癌患者术后总生存期较短(P<0.05)。结论含有HNRNPC的EVs影响子宫颈癌的生长和转移,并促进HUVECs细胞的血管生成,从而加速子宫颈癌的进展。此外,子宫颈癌组织中HNRNPC高表达与患者较差的预后相关,表明含�Purpose To investigate the predictive value of the methylation regulator heterogeneous nuclear ribonucleoprotein C(HNRNPC)in the extracellular vesicles(EVs)derived from cervical cancer cells for tumor invasion and metastasis.Methods In vitro experiments were conducted using a human normal cervical epithelial cell line(HcerEpic),three cervical cancer cell lines(HeLa,SiHa and CaSki)and human umbilical vein endothelial cells(HUVECs).HNRNPC protein expression was assessed by Western blot.CaSki cells were transfected with either sh-NC or sh-HNRNPC,and EVs were subsequently isolated from culture supernatant.The effects of EVs on the proliferation,migration and invasion of SiHa cells,as well as on angiogenesis in HUVECs,were investigated respectively.For tissue microarray analysis,normal cervical tissues(n=8),low-grade squamous intraepithelial lesions(LSIL,n=32)and high-grade squamous intraepithelial lesions(HSIL,n=37),and cervical carcinoma specimens(n=153)were obtained from our institution.Based on HNRNPC immunoscores,cervical cancer patients were divided into two groups:high(n=99)and low(n=54)HNRNPC expression group,and their clinicopathological features and prognosis were compared.Results Compared with HcerEpic cells,the expression of HNRNPC increased in SiHa,HeLa and CaSki cells.In SiHa cells treated with EVs from CaSki cell,the group receiving EVs from sh-HNRNPC-transfected CaSki cells showed significantly reduced proliferation,colony formation,migration,and invasion compared with the sh-NC-EVs group(P<0.05).Similarly,in HUVECs treated with CaSki-EVs,compared with sh-NC-EVs group,the sh-HNRNPC-EVs group demonsteated significantly lower protein expression of PCNA and VEGFA and reduced angiogenesis length(P<0.05).HNRNPC expression gradually increased during the transformation of cervical epithelial cells(F=106.9,P<0.001),and was significantlyelevated in HSIL and cervical cancer tissues compared with normal tissues(P<0.001).Moreover,in cervical cancer,high HNRNPC expression was significantly related to poorer cellula

关 键 词：子宫颈肿瘤 细胞外囊泡 核内不均一核糖核蛋白C 侵袭转移 

分 类 号：R737.33[医药卫生—肿瘤]