长链非编码RNA浆细胞瘤变异性易位基因1参与盆腔器官脱垂的机制  

Mechanism of long non-coding RNA plasmacytoma variant translocation 1 involved in pelvic organ prolapse

作　　者：胡新明 乔艳华 王肖帆 李林玉 赵冰[1] Hu Xinming;Qiao Yanhua;Wang Xiaofan;Li Linyu;Zhao Bing(Department of Gynecology and Obstetrics,The Third Affiliated Hospital of Zhengzhou University,Zhengzhou 450000,Henan Province,China;Academic Affairs,Zheng Zhou Feng Yang Foreign Language School,Zhengzhou 450000,Henan Province,China)

机构地区：[1]郑州大学第三附属医院妇产科,河南省郑州市450000 [2]郑州枫杨外国语学校教务处,河南省郑州市450000

出　　处：《中国组织工程研究》2023年第5期669-675,共7页Chinese Journal of Tissue Engineering Research

基　　金：河南省高校科技创新人才支持计划(18HASTIT045),项目负责人:赵冰。

摘　　要：背景:作者所在课题组前期通过高通量测序结果发现长链非编码RNA浆细胞瘤变异性易位基因1在盆腔器官脱垂患者子宫主韧带中表达明显降低,转化生长因子β1信号通路也是相关通路之一。目的:观察长链非编码RNA浆细胞瘤变异性易位基因1对盆底韧带成纤维细胞的影响,探讨其在盆腔器官脱垂疾病发病机制中的可能作用。方法:提取大鼠盆底韧带组织标本,分离成纤维细胞,进行原代培养。慢病毒构建长链非编码RNA浆细胞瘤变异性易位基因1干扰和过表达载体加入成纤维细胞中。通过免疫荧光、MTT、细胞划痕实验、流式细胞术来检测成纤维细胞的形态、增殖、迁移和凋亡情况;RT-PCR和Western-blot检测转化生长因子β1、smad2/3、c-Myc的mRNA及蛋白表达情况。结果与结论:①长链非编码RNA浆细胞瘤变异性易位基因1过表达时可显著促进成纤维细胞的增殖,提高细胞的迁移能力,抑制成纤维细胞凋亡;长链非编码RNA浆细胞瘤变异性易位基因1干扰时结果相反;②此外,长链非编码RNA浆细胞瘤变异性易位基因1过表达时细胞中的c-Myc、转化生长因子β1、smad2/3的mRNA和蛋白相对表达量均明显增加,其干扰时则显著抑制c-Myc和转化生长因子β1的表达;③提示长链非编码RNA浆细胞瘤变异性易位基因1能够影响成纤维细胞的形态和生物学功能,可能与转化生长因子β1/smad2/3/c-Myc通路有关;该研究可为盆腔器官脱垂患者的临床诊断和治疗提供新思路。BACKGROUND:Previous studies have shown that the expression of long non-coding RNA plasmacytoma variant translocation 1(LncRNA PVT1)was significantly reduced in the uterine cardinal ligament of patients with pelvic organ prolapse through high-throughput sequencing.Transforming growth factorβ1 signaling pathway is also one of the related pathways.OBJECTIVE:To investigate the effect of LncRNA PVT1 on pelvic floor ligament fibroblasts and its possible role in the pathogenesis of pelvic organ prolapse.METHODS:Tissue samples of the pelvic floor ligament were extracted from rats and fibroblasts were isolated for primary culture.LncRNA PVT1 interference and overexpression vectors were constructed by lentivirus and added into fibroblasts.Morphology,proliferation,migration and apoptosis of fibroblasts were detected by immunofluorescence,MTT,cell scratch assay and flow cytometry test respectively.The mRNA and protein expressions of transforming growth factor-β1,Smad2/3 and c-Myc were detected by RT-PCR and western blot respectively.RESULTS AND CONCLUSION:Overexpression of LncRNA PVT1 could significantly promote the proliferation of fibroblasts,improve the migration ability of cells,and inhibit the apoptosis of fibroblasts.The results of LncRNA PVT1 interference were opposite.In addition,the mRNA and protein relative expressions of c-Myc,transforming growth factor-β1 and Smad2/3 were significantly increased when LncRNA PVT1 gene was overexpressed,while the expressions of c-Myc and transforming growth factor-β1 were significantly inhibited when LncRNA PVT1 gene was interfered.To conclude,LncRNA PVT1 can affect the morphology and biological function of fibroblasts,which may be related to the transforming growth factor-β1/Smad2/3/c-Myc pathway.This study provides new ideas for the clinical diagnosis and treatment of pelvic organ prolapse.

关 键 词：长链非编码RNA 浆细胞瘤变异性易位基因1 盆腔器官脱垂 成纤维细胞 过表达 干扰 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R711.2