Song Huicong, Sun Xuefeng, Wang Xiaohua, Xie Tianhai, Zheng Zhihui, Ji Ying, Cui Yanyan
Department of Oncology, Affiliated Hospital of Chifeng University, Chifeng, Inner Mongolia, China.
Cell Biochem Biophys. 2025 Jun;83(2):2047-2058. doi: 10.1007/s12013-024-01615-z. Epub 2024 Nov 27.
The medial overall survival is low in patients with gastric cancer (GC) at advanced stage, in which drug resistance plays an important role. β-elemene has been established as the suppressed role on GC cell proliferation, however, the concrete mechanism of it remains unclear in cisplatin (DDP)-resistance GC. Cell counting kit-8 (CCK8) assay was used to measure the half maximal inhibitory concentration (IC) values of DDP in DDP-resistance GC cell lines. Cell apoptotic rates and invasive ability were tested by flow cytometry and transwell assay. Western blot and reverse-transcription quantitative polymerase chain reaction (RT-qPCR) were utilized to detect the protein and mRNA levels of methyltransferase like-3 (METTL3) and ADP ribosylation factor 6 (ARF6). SRAMP websites and methylated RNA immunoprecipitation (MeRIP) assay were applied to predicted m6A sites and verified m6A levels of ARF6 respectively. RNA immunoprecipitation (RIP) was used to explore the interaction between these two molecules. Xenograft tumor models were constructed to demonstrate the effects of β-elemene in vivo. β-elemene improved drug sensitivity and curbed malignant cell activities of DDP-resistance GC cells in vitro. ARF6 was upregulated in DDP-resistance GC cells and tissues, and its overexpression could abrogate the effects on DDP-resistant GC cells mediated by β-elemene treatment. Intracellular and exosomal METLL3 expression were elevated in and from DDP-resistance GC cell lines. Exosomal METTL3 released from DDP-resistance GC cells could counteract the effects of β-elemene on DDP-resistance GC cells partly via regulating ARF6 expression in the m6A-dependent manner. β-elemene could suppress DDP-resistance tumor growth in vivo. In conclusion, β-elemene could repress tumor growth and drug resistance via exosomal METTL3-m6A-ARF6 axis.
晚期胃癌（GC）患者的总体中位生存期较低，其中耐药性起重要作用。β-榄香烯已被证实对GC细胞增殖具有抑制作用，然而，其在顺铂（DDP）耐药GC中的具体机制仍不清楚。采用细胞计数试剂盒-8（CCK8）法检测DDP在DDP耐药GC细胞系中的半数最大抑制浓度（IC）值。通过流式细胞术和Transwell实验检测细胞凋亡率和侵袭能力。采用蛋白质免疫印迹法（Western blot）和逆转录定量聚合酶链反应（RT-qPCR）检测甲基转移酶样3（METTL3）和ADP核糖基化因子6（ARF6）的蛋白质和mRNA水平。分别应用SRAMP网站和甲基化RNA免疫沉淀（MeRIP）实验预测ARF6的m6A位点并验证其m6A水平。采用RNA免疫沉淀（RIP）实验探究这两种分子之间的相互作用。构建异种移植瘤模型以证明β-榄香烯在体内的作用。β-榄香烯在体外提高了DDP耐药GC细胞的药物敏感性并抑制了恶性细胞活性。ARF6在DDP耐药GC细胞和组织中上调，其过表达可消除β-榄香烯处理对DDP耐药GC细胞的影响。DDP耐药GC细胞系内和外泌体中的METTL3表达均升高。DDP耐药GC细胞释放的外泌体METTL3可部分通过以m6A依赖的方式调节ARF6表达来抵消β-榄香烯对DDP耐药GC细胞的影响。β-榄香烯在体内可抑制DDP耐药肿瘤的生长。总之，β-榄香烯可通过外泌体METTL3-m6A-ARF6轴抑制肿瘤生长和耐药性。
