肿瘤源性LINC01006经外泌体途径促进前列腺癌增殖侵袭  

Tumor-derived LINC01006 promotes proliferation and invasion of prostate cancer via exosome-mediated pathway

作　　者：王世远 姜辰一 赵福军 WANG Shi-yuan;JIANG Chen-yi;ZHAO Fu-jun(Department of Urology,Shanghai General Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200080,China)

机构地区：[1]上海交通大学医学院附属第一人民医院泌尿外科,上海200080

出　　处：《川北医学院学报》2025年第4期409-415,共7页Journal of North Sichuan Medical College

基　　金：国家自然科学基金(82371634)。

摘　　要：目的:明确LINC01006在去势抵抗性前列腺癌中的差异性表达,证明LINC01006介导前列腺癌细胞增殖侵袭性增强。探讨LINC01006包装至外泌体并在细胞间传递,使PCa细胞增殖侵袭性增强。方法:分析正常前列腺组织和前列腺癌组织外泌体差异表达的转录组,并进行筛选。验证LINC01006在正常前列腺组织和癌组织中的表达差异。利用构建shRNA表达载体质粒,并结合慢病毒包被技术,通过转染LNCaP及22RV1细胞。验证LINC01006被被膜结构覆盖。高超速离心法提取PCa细胞外泌体并以Western blot验证。RT-qPCR技术验证LINC01006在外泌体中的表达差异。CCK-8增殖活性实验及平板克隆实验来进行验证LINC01006敲除的情况下,PCa细胞增殖、迁移能力的变化情况。CCK-8增殖活性实验及平板克隆实验同种、异种PCa细胞来源的外泌体共培养时,LINC01006敲除的情况下PCa细胞增殖、迁移能力恢复情况。结果:相比较正常前列腺上皮细胞(RWPE-1),LNCaP和22RV1细胞中的LINC01006的表达上调。敲减的LNCaP与对照组相比,LINC01006的表达下调了84.7%(P<0.01),敲减的22RV1与对照组相比,LINC01006的表达下调了80.3%(P<0.01)。CM+RNase+T组相比另外两组,LINC01006的检测水平显著降低(P<0.01)。高超速离心法成功提取外泌体,特异性蛋白CD63,CD81,HSP70高表达(P<0.05)。源于LNCaP细胞和源于22RV1细胞的外泌体LINC01006的表达水平均显著升高(P<0.01)。与对照组(siCtrl)相比,敲除组(si1006)细胞在第3、4天的吸光度显著低于对照组(P<0.01)。与对照组(siCtrl)相比,敲除组的细胞克隆形成数显著减少(P<0.05)。与对照组(Ctrl)相比,外泌体组(LNCaP exosome)细胞在第4天的吸光度显著高于对照组(P<0.05)。与对照组(Ctrl)相比,外泌体组的细胞克隆形成数显著增多(P<0.05)。与对照组(Ctrl)相比,外泌体组(22RV1 exosome)细胞在第3、4天的吸光度显著高于对照组(P<0.05)。与对照组(Ctrl)相比,外泌体组的Objective:To clarify the differential expression of LINC01006 in Castration-Resistant Prostate Cancer.To prove that LINC01006 mediates the enhanced proliferation and invasiveness of prostate cancer cells.To clarify that LINC01006 is packaged into exosomes and delivered between cells,resulting in enhanced PCa cell proliferation and invasiveness.Methods:Screen the differentially expressed transcriptomes of normal prostate tissue and prostate cancer tissue by analyzing the biological information database.The expression difference of LINC01006 in normal prostate tissue and cancer tissue was verified by RT-qPCR technology.Using the construction of shRNA expression vector plasmid,combined with lentivirus coating technology,LNCaP and 22RV1 cells were transfected.Using the technology of extracting extracellular medium,by adding RNase and changing the permeability of the probe,it was verified that LINC01006 was covered by the membrane structure.PCa cell exosomes were extracted by high-speed ultracentrifugation and verified by Western Blot.The expression difference of LINC01006 in exosomes was verified by RT-qPCR technology.CCK-8 assay and colony formation assay were used to verify the changes of PCa cell proliferation and migration ability in the case of LINC01006 knockout.CCK-8 proliferation activity experiment and plate cloning experiment When exosomes derived from homologous and xenogeneic PCa cells were co-cultured,the recovery of PCa cell proliferation and migration ability in the case of LINC01006 knockout.Results:Compared with normal prostate epithelial cells(RWPE-1),the expression of LINC01006 in LNCaP and 22RV1 cells was up-regulated(P<0.01).The expression of LINC01006 in knockdown LNCaP was down-regulated by 84.7%compared with the control group(P<0.01),and the expression of LINC01006 in knock-down 22RV1 was down-regated by 80.3%compared with the control group(P<0.01).There was no significant difference between CM group and CM+RNase group(P>0.05).Compared with the other two groups,the detection of LINC01006 in th

关 键 词：前列腺癌 长链非编码RNA 外泌体 细胞增殖 

分 类 号：R737.25[医药卫生—肿瘤]