摘要
目的:探讨石上柏外泌体样纳米囊泡对肝癌细胞的影响。方法:采取改良超高速离心法与蔗糖密度梯度离心法提取并纯化石上柏外泌体样纳米囊泡,应用透射电镜与纳米颗粒跟踪分析技术(NTA)进行鉴定。采用绿色荧光探针(DIO)标记石上柏外泌体样纳米囊泡并观察其被细胞摄取的情况。采用CCK-8法检测不同浓度的石上柏外泌体样纳米囊泡对肝癌细胞活力的影响,并计算对HepG2细胞的半数抑制浓度(IC_(50))值。观察石上柏外泌体样纳米囊泡对不同肝癌细胞形态变化的影响。采用克隆集落实验、划痕迁移实验、Transwell侵袭实验、流式细胞术探究石上柏外泌体样纳米囊泡对肝癌细胞增殖、迁移、侵袭、凋亡的影响。结果:经证实提取物为外泌体样纳米囊泡,石上柏外泌体样纳米囊泡能明显抑制肝癌细胞活力,抑制肝癌细胞的增殖、迁移和侵袭,促进细胞凋亡(P<0.05)。结论:石上柏外泌体样纳米囊泡对肝癌细胞有一定抑制性,且可能与顺铂协同发挥抑制和促凋亡作用。
Objective:To investigate the effects of exosome-like nanovesicles from Selaginella doederleinii Hieron (SDHENs) on hepatocellular carcinoma cells.Methods:The refined ultra-high speed centrifugation method and sucrose density gradient centrifugation method were used to extract and purify SDHENs.It was identified by transmission electron microscopy (TEM) and nanoparticle tracking analysis (NTA).Green fluorescent probe (DIO) was used to label SDHENs,observing the uptake of SDHENs by cells.CCK-8 method was used to detect the effect of different concentrations of SDHENs on hepatocellular carcinoma cells,and the half inhibitory concentration (IC_(50)) of HepG2 cells was calculated.The effect of SDHENs on the morphological changes of different hepatocellular carcinoma cells was observed.The effects of SDHENs on the proliferation,migration,invasion and apoptosis of hepatocellular carcinoma cells were observed by colony-formation assay,wound healing,Transwell invasion assay and flow cytometry.Results:It was proved that the extracts were exosome-like nanovesicles,and the SDHENs could significantly inhibit the viability of hepatocellular carcinoma cells,inhibit the proliferation,migration and invasion of hepatocellular carcinoma cells,and promote cell apoptosis(P<0.05).Conclusion:The SDHENs have certain inhibitory effect on hepatocellular carcinoma cells,and may cooperate with cisplatin to play an inhibitory and pro-apoptotic role.
作者
胡炬
梁子成
谭小宁
周芳
曾普华
HU Ju;LIANG Zicheng;TAN Xiaoning;ZHOU Fang;ZENG Puhua(Hunan University of Chinese Medicine,Hunan Changsha 410208,China;Hunan Provincial Hospital of Integrated Traditional Chinese and Western Medicine,Hunan Changsha 410006,China;Cancer Institute of Hunan Academy of Traditional Chinese Medicine,Hunan Changsha 410006,China)
出处
《现代肿瘤医学》
CAS
2024年第16期2937-2944,共8页
Journal of Modern Oncology
基金
国家中医药管理局首批全国青年岐黄学者人才项目(编号:国中医药人教发【2020】7号)
湖南省重点研发项目(编号:2021SK2006,2023SK2057)
湖南省自然基金项目(编号:2021JJ30417,2023JJ40403)
湖南中医药大学研究生创新课题(编号:2023CX113)
湖南省中医药研究院科研课题(编号:202011)。
关键词
肝癌
石上柏
外泌体
中医药
hepatocellular carcinoma
Selaginella doederleinii Hieron
exosome
traditional Chinese medicine
