不同腹膜后淋巴结转移能力的卵巢癌细胞小RNA(miRNA)表达差异  

Differential expression of microRNAs(miRNA)in retroperitoneal lymph node metastasis of ovarian carcinoma cells

作　　者：杨梅琳[1] 康玉[1,2,3] 洪珊珊[1] 罗建民[4] 徐丛剑[1,2,3,5] 

机构地区：[1]复旦大学附属妇产科医院妇科,上海200011 [2]复旦大学上海医学院妇产科学系,上海200011 [3]上海市女性生殖内分泌相关疾病重点实验室,上海200011 [4]复旦大学附属肿瘤医院核医学科,上海200032 [5]复旦大学生物医学研究院,上海200032

出　　处：《复旦学报（医学版）》2014年第4期476-482,共7页Fudan University Journal of Medical Sciences

基　　金：上海市自然科学基金(13ZR1404300);上海市科委实验动物专项基金资助项目(13140901500)~~

摘　　要：目的筛选具有腹膜后高淋巴结转移能力的浆液性卵巢癌细胞株,并分析其导致淋巴高转移的相关小RNA(microRNA,miRNAs)。方法接种人卵巢癌细胞株SKOV-3于BALB/c裸鼠腹腔,从腹膜后淋巴结中筛选出卵巢癌细胞,再次接种于裸鼠腹腔,重复分选4次后获得了稳定的高腹膜后淋巴结转移细胞株SKOV-3/LN403。对比SKOV-3和SKOV-3/LN403细胞的形态、增殖和侵袭能力,应用miRCURYTMLNA Array技术对两者的miRNA进行差异分析,通过real-time PCR验证差异表达的miRNAs。结果稳定筛选的腹膜后高淋巴结转移卵巢癌细胞株SKOV-3/LN403,淋巴转移率高达90%(n=10),并能成功模拟卵巢癌淋巴转移患者的临床表现。SKOV-3/LN403具有比SKOV-3更强的增殖(P<0.05)和侵袭能力(P<0.01)。miRNA的表达谱提示14种miRNA表达差异明显,包括3种miRNA上调(hsa-miR-886-5p,hsa-miR-640,hsa-miR-801),11种miRNA下调(hsa-let-7a,hsa-miR-30a,hsa-miR-491-3p,hsa-let-7i,hsa-miR-125b-1,hsa-miR-27a,hsa-miR-24,hsalet-7b,hsa-miR-16,hsa-miR-20a,hsa-miR-26b)。Real time PCR结果和芯片结果一致。结论 SKOV-3/LN403可用于构建卵巢癌腹膜后淋巴结转移模型,其高腹膜后淋巴结转移力与miRNA分子的调控相关,为卵巢癌腹膜后淋巴结转移机制提供潜在研究靶点。Objective To establish a human serous ovarian carcinoma cell line with high potential for retroperitoneal lymph node metastasis,and to study its differential expression of microRNAs （miRNA） between ovarian carcinoma cells with high and low potentials for lymphatic metastasis.Methods Ovarian carcinoma cells,SKOV-3,were inoculated into the abdominal cavities of BALB/c nude mice.Tumor cells in the retroperitoneal lymph node were isolated and cultured in vitro.Then we inoculated these isolated cancer cells into the abdominal cavities of BALB/c nude mice again.By repeating this procedure 4 times,we obtained the human ovarian cancer （OC） cell line SKOV-3/LN403 with high potential for spontaneous lymph node metastasis.The biologic characteristics of SKOV-3 and SKOV-3/ LN403 were compared with regard to cell morphology,cell proliferation assays,and cell invasion in vitro,miRNAs with differential expression were demonstrated between SKOV-3 and SKOV-3/LN403 by miRCURY LNATM microRNA array.Finally,real-time PCR was used to validate differentially expressed miRNAs.Results The lymph node metastasis rate in SKOV-3/LN403-inoculated mice was 90% （n =10）,and it also could modify the clinical manifestation of ovarian cancer.SKOV-3/ LN403 showed higher reproductive activity （P〈0.05） and invasiveness （P〈0.01） than SKOV-3.The results of the miRNA array indicated 3 miRNAs were up-regulated （hsa-miR-886-5p,hsa-miR-640,hsa-miR-801） and 11 miRNAs were down-regulated significantly （hsa-let-7a,hsa-miR-30a,hsa-miR491-3p,hsa-let-7i,hsa-miR-125b-1,hsa-miR-27a,hsa-miR-24,hsa-let-7b,hsa-miR-16,hsa-miR-20a,hsa-miR-26b）.The real-time PCR results were consistent with the miRNA array.Conclusions The SKOV-3/LN403 could be successfully used to establish an ovarian cancer model with spontaneous retroperitoneal lymph node metastasis.Differential expression of miRNAs might be attributed to its high potential for lymph node metastasis.This cell line provided a pre-research platform for the molecular mechanisms of OC lymph

关 键 词：卵巢癌 浆液性卵巢癌 腹膜后淋巴结 淋巴转移 小microRNA (miRNA) 

分 类 号：R737.31[医药卫生—肿瘤]