提取物通过靶向 PDCD4 和 EIF3H 减少胃癌干细胞的干性。
Extract Reduces Stemness of Gastric Cancer Stem Cells by Targeting PDCD4 and EIF3H.
机构信息
Medical University of Anhui, Anhui, China.
出版信息
Integr Cancer Ther. 2021 Jan-Dec;20:15347354211058168. doi: 10.1177/15347354211058168.
BACKGROUND
ethyl acetate extract (COE) has shown a strong anti-gastric cancer effect, but the understanding of its mechanism is still lacking. The results of previous studies indicated that COE may be able to inhibit the stemness of gastric cancer stem cells (GCSCs) by regulating PDCD4 and EIF3H expression.
AIMS
To explore if COE could inhibit the stemness of GCSCs by regulating PDCD4 and EIF3H expression in vitro and in vivo.
PROCEDURE
The GCSCs model was established by stem cell-conditioned culture. Spheroid formation and flow cytometry assays were used to detect the effect of COE on the spheroid formation ability of GCSCs and the percentage of CD44/CD24 and ALDH cell subpopulations. Western blot analysis was applied to measure the expression of GCSCs biomarkers (Nanog, Oct-4, and SOX-2), PDCD4, and EIF3H in GCSCs treated with COE; and RT-PCR was performed to investigate the effect of COE on PDCD4 mRNA expression in GCSCs. An in vivo tumorigenicity experiment was also conducted to evaluate the effect of COE on tumor-initiating ability of GCSCs in vivo; and the expression of PDCD4 and EIF3H in xenograft tissues was examined by immunohistochemistry (IHC) staining.
RESULTS
After culture in stem cell-conditioned medium, SGC7901 cells manifested significantly enhanced spheroid formation ability, upregulated Nanog, Oct-4, and SOX-2 expression and increased percentages of CD44/CD24 and ALDH cell subpopulations, indicating successful establishment of the GCSCs model. COE treatment significantly inhibited the spheroid formation ability of GCSCs and reduced the percentage of CD44/CD24 and ALDH cell subpopulations. The western blot analysis showed a significant decrease of Nanog, Oct-4, SOX-2, and EIF3H expression and an increase of PDCD4 expression in GCSCs after COE treatment in a concentration-dependent manner. COE treatment also significantly upregulated the mRNA expression of PDCD4 in GCSCs. In addition, COE displayed a strong inhibitory effect on the tumor-initiating ability of GCSCs in vivo and upregulated PDCD4 and downregulated EIF3H expression in xenograft tissues.
CONCLUSION
COE may be able to inhibit GC growth by suppressing the stemness of GCSCs via regulating PDCD4 and EIF3H expression.
背景
乙酸乙酯提取物（COE）已显示出很强的抗胃癌作用，但对其作用机制的了解仍不够充分。先前的研究结果表明，COE 可能通过调节 PDCD4 和 EIF3H 的表达来抑制胃癌干细胞（GCSCs）的干性。
目的
在体外和体内探索 COE 是否可以通过调节 PDCD4 和 EIF3H 的表达来抑制 GCSCs 的干性。
方法
通过干细胞条件培养建立 GCSCs 模型。采用球体形成和流式细胞术检测 COE 对 GCSCs 球体形成能力和 CD44/CD24 和 ALDH 细胞亚群比例的影响。Western blot 分析用于检测 COE 处理后 GCSCs 中 GCSCs 标志物（Nanog、Oct-4 和 SOX-2）、PDCD4 和 EIF3H 的表达；并通过 RT-PCR 检测 COE 对 GCSCs 中 PDCD4 mRNA 表达的影响。还进行了体内肿瘤形成实验，以评估 COE 对体内 GCSCs 起始肿瘤能力的影响；并通过免疫组织化学（IHC）染色检测异种组织中 PDCD4 和 EIF3H 的表达。
结果
在干细胞条件培养基中培养后，SGC7901 细胞表现出明显增强的球体形成能力，上调 Nanog、Oct-4 和 SOX-2 的表达，并增加 CD44/CD24 和 ALDH 细胞亚群的比例，表明成功建立了 GCSCs 模型。COE 处理显著抑制 GCSCs 的球体形成能力，并降低 CD44/CD24 和 ALDH 细胞亚群的比例。Western blot 分析显示，COE 处理以浓度依赖性方式显著降低 GCSCs 中 Nanog、Oct-4、SOX-2 和 EIF3H 的表达，并增加 PDCD4 的表达。此外，COE 处理还显著上调 GCSCs 中 PDCD4 的 mRNA 表达。此外，COE 在体内对 GCSCs 的起始肿瘤能力具有很强的抑制作用，并上调异种组织中的 PDCD4 和下调 EIF3H 的表达。
结论
COE 可能通过调节 PDCD4 和 EIF3H 的表达来抑制 GC 生长，从而抑制 GCSCs 的干性。
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