Department of General Surgery, Nanjing Tongren Hospital, No. 2007, Jiyin Avenue, Jiangning District, Nanjing, 211102, China.
Mol Biotechnol. 2024 Jun;66(6):1389-1401. doi: 10.1007/s12033-023-00795-y. Epub 2023 Jun 23.
Human umbilical cord mesenchymal stem cell (hucMSC)-derived exosomes (Exo) have been frequently investigated for disease control. This study was designed to explore the effects of hucMSC-Exo carrying lncRNA family with sequence similarity 99-member B (Exo-lncRNA FAM99B) on hepatocellular carcinoma (HCC) cell behaviour. The expression of lncRNA FAM99B in HCC cells was measured by reverse-transcription quantitative polymerase chain reaction. Protein levels of exosomal markers were quantified using western blotting. Flow cytometry analyses were performed to detect surface markers of hucMSCs and to measure the effects of Exo-lncRNA FAM99B on HCC cell cycle progression and cell apoptosis. Nanoparticle tracking analysis was used to measure the particle size of the exosomes. Additionally, cell viability was evaluated using methyl thiazolyl tetrazolium assays, and Transwell assays were performed to measure cell migration and invasion. Xenograft tumor models were established to explore the role of Exo-lncRNA FAM99B in vivo. Experimental results revealed that lncRNA FAM99B was downregulated in HCC cell lines, and low level of FAM99B is associated with poor survival rates in patients with HCC according to bioinformatics analysis. HucMSCs were identified in a good morphology with positively expressed CD105, CD29, and CD44 as well as negatively expressed CD31, CD14, and HLA-DR. High protein levels of exosomal markers (Alix, CD63 and TSG101) identified the existence of HucMSC-Exo. Importantly, the hucMSCs-Exo could enter HCC cells and exerted a suppressive effect on malignant cell activities. Moreover, overexpression of Exo-lncRNA FAM99B enhanced cell cycle arrest and cell apoptosis while suppressing cell viability, migration, and invasion in HCC. Exo-siRNA-FAM99B exerted the opposite effects on HCC cell process. In vivo experiments verified that Exo-lncRNA FAM99B inhibited tumorigenesis in HCC. In summary, lncRNA FAM99B derived from hucMSC-Exo inhibited malignant cellular phenotypes and tumorigenesis in HCC, which might provide a novel therapeutic strategy for HCC treatment.
人脐带间充质干细胞（hucMSC）衍生的外泌体（Exo）经常被用于疾病控制的研究。本研究旨在探讨携带长链非编码 RNA 家族与序列相似性 99 成员 B（Exo-lncRNA FAM99B）的 hucMSC-Exo 对肝细胞癌（HCC）细胞行为的影响。采用逆转录定量聚合酶链反应测定 HCC 细胞中 lncRNA FAM99B 的表达。采用 Western blot 定量测定外泌体标志物的蛋白水平。采用流式细胞术分析检测 hucMSC 的表面标志物，并检测 Exo-lncRNA FAM99B 对 HCC 细胞周期进程和细胞凋亡的影响。采用纳米颗粒跟踪分析测定外泌体的粒径。此外，采用噻唑蓝比色法评估细胞活力，采用 Transwell 测定评估细胞迁移和侵袭。建立异种移植肿瘤模型以探讨 Exo-lncRNA FAM99B 在体内的作用。实验结果显示，lncRNA FAM99B 在 HCC 细胞系中下调，且生物信息学分析显示 FAM99B 低表达与 HCC 患者生存率降低相关。hucMSC 形态良好，阳性表达 CD105、CD29 和 CD44，阴性表达 CD31、CD14 和 HLA-DR。外泌体标志物（Alix、CD63 和 TSG101）的高蛋白水平鉴定出 hucMSC-Exo 的存在。重要的是，hucMSC-Exo 可以进入 HCC 细胞，并对恶性细胞活性发挥抑制作用。此外，过表达 Exo-lncRNA FAM99B 增强 HCC 细胞的细胞周期阻滞和细胞凋亡，同时抑制细胞活力、迁移和侵袭。Exo-siRNA-FAM99B 对 HCC 细胞过程产生相反的影响。体内实验证实 Exo-lncRNA FAM99B 抑制 HCC 肿瘤发生。总之，hucMSC-Exo 来源的 lncRNA FAM99B 抑制 HCC 中恶性细胞表型和肿瘤发生，为 HCC 治疗提供了一种新的治疗策略。
