沉默NAC-1基因对人卵巢癌耐药细胞株SKOV3/DDP裸鼠移植瘤化疗敏感的影响  

The effect of silencing NAC-1 gene on chemotherapy sensitivity of human ovarian cancer drug resistant cell line SKOV3/DDP transplanted tumor in nude mice

作　　者：李冬冬 王莉[1] 钟洁 凌晨祁 韩英 Li Dongdong;Wang Li;Zhong Jie(Department of Gynecology,Shidong Hospital,University of Shanghai for Science and Technology,Shanghai 200438)

机构地区：[1]上海理工大学附属市东医院妇科,上海200438

出　　处：《现代妇产科进展》2024年第2期81-86,共6页Progress in Obstetrics and Gynecology

基　　金：上海市杨浦区卫生和计划生育委员会科研课题(No:YPQ 202104,No:YP18Q03)。

摘　　要：目的:探讨核转录因子NAC-1基因沉默对人卵巢癌耐药细胞株SKOV3/DDP裸鼠移植瘤化疗敏感性的影响。方法:采用LV4-LUC-GFP慢病毒转染建立NAC-1基因沉默SKOV3/DDP细胞株,将细胞接种到免疫缺陷小鼠皮下,建立人卵巢癌耐药细胞株SKOV3/DDP裸鼠移植瘤模型,给予顺铂化疗,比较NAC-1基因沉默对SKOV3/DDP在裸鼠体内增殖的影响。取肿瘤组织进行转录组测序,利用Illumina平台进行转录组mRNA测序,筛选差异表达基因,并进行GO和KEGG功能注释和富集分析,揭示NAC-1基因影响移植瘤化疗敏感的可能分子机制。使用STRING数据库构建DEGs编码蛋白互作网络,筛选MCC算法、Degree算法、Closeness算法3种算法共有基因作为关键基因,使用Metascape网站在线分析这些共同基因的基因功能。结果:与对照组相比,抑制NAC-1基因后裸鼠皮下肿瘤体积变小,重量减轻,差异有统计学意义(P<0.05)。转录组学分析显示,抑制NAC-1基因导致肿瘤组织中460个基因显著下调,568个基因显著上调。差异基因(DEGs)在多项GO富集类别中具有显著差异,如细胞代谢、有丝分裂、坏死、炎症、信号传递等。KEGG功能富集结果显示,NAC-1基因沉默可能影响了细胞因子-细胞因子受体相互作用、白细胞跨内皮迁移、神经活性配体-受体相互作用、细胞黏附分子、坏死、ECM受体相互作用、PI3K-Akt信号通路、NF-κB信号通路等。筛选获得的关键基因主要富集在白细胞介素的信号传导、粒细胞-巨噬细胞集落刺激因子(GM-CSF)通路、炎症反应等通路。结论:抑制NAC-1基因能提高人卵巢癌耐药细胞株SKOV3/DDP化疗敏感性,其可能通过多个信号通路影响肿瘤细胞的代谢、增殖、死亡、炎症反应等。Objective:To investigate the effect of nuclear transcription factor NAC-1 gene silencing on chemotherapy sensitivity of a nude mouse transplanted tumor model constructed from multi-drug resistant human ovarian cell line SKOV3/DDP.Methods:The NAC-1 gene silenced SKOV3/DDP cell line was transfected with LV4-LUC-GFP lentivirus,and then the cells were inoculated subcutaneously into immunodeficient mice to establish a human ovarian cancer resistant SKOV3/DDP nude mouse model of transplanted tumor.The cells were then subcutaneously inoculated into immunodeficient mice to establish a nude mouse transplant tumor model of human ovarian cancer resistant cell line SKOV3/DDP.Tumor-bearing nude mice was treated with cisplatin chemotherapy to compare the effect of silencing the NAC-1 gene on the proliferation of SKOV3/DDP in vivo.Further,the transcriptome of tumor tissue was sequenced,and the transcriptome mRNA was sequenced using the illumina platform to screen differentially expressed genes.GO and KEGG functional annotation and enrichment analysis were performed to reveal the possible molecular mechanism of NAC-1 gene affecting the chemosensitivity of transplanted tumor.Then,a DEGs encoded protein interaction network was constructed using the STRING database and the common genes shared by the MCC algorithm,Degree algorithm and Closeness algorithm were screened as key genes.The functions of these common genes were analyzed online using the Metascape website.Results:Compared with the control group,inhibiting the NAC-1 gene reduced the volume and weight of subcutaneous tumors in nude mice,with significant differences in results.Transcriptome analysis showed that inhibition of NAC-1 gene resulted in 460 genes being significantly down regulated and 568 genes being significantly up regulated in tumors.Differential genes(DEG)had significant differences in many GO enrichment categories,such as cell metabolism,mitosis,necrosis,inflammation,signal transmission,etc.KEGG function enrichment results showed that NAC-1 gene silencing may a

关 键 词：NAC-1 卵巢癌细胞 顺铂耐药 转录组分析 

分 类 号：R737.31[医药卫生—肿瘤]