Zhou Yang, Zhuang Haoran, Liu Yuxiang, Yin Jing, Wei Xiaoying, Qiu Yue, Tian Zhen, Miao Tingyu, Chen Jing, Li Peifen, Xu Xiao, Wu Wenjuan, Li Huanan, Shen Weigan
Department of Cell Biology, School of Medicine of Yangzhou University, Yangzhou, Jiangsu, China.
Department of Oncology, Taizhou Hospital of Traditional Chinese Medicine, Taizhou, Jiangsu, China.
Discov Oncol. 2023 Dec 18;14(1):233. doi: 10.1007/s12672-023-00838-5.
Celastrol has been revealed to exhibit anticancer pharmacological activity, however, the molecular mechanisms of celastrol involved in pancreatic cancer remain to be further elucidated. The present study was to illustrate whether celastrol suppresses pancreatic cancer through modulating RNA mA modification.
Effect of celastrol treatment on the malignant phenotypes of pancreatic cancer cells was evaluated by CCK-8 assay, EdU assay, colony formation assay, flow cytometry analysis and subcutaneous xenograft experiments. RNA sequencing (RNA-seq) analysis was carried out to analyze the genes differentially expressed in celastrol-treated pancreatic cancer cells. RT-qPCR, Western blotting and immunohistochemistry were employed to evaluate the expression of the indicated genes. RNA dot blot and quantification of total RNA mA modification assays, MeRIP-qPCR assay, RIP-qPCR assay, RNA stability and protein stability assays were applied to evaluate the regulatory mechanism of celastrol treatment in pancreatic cancer cells.
We demonstrated that celastrol suppressed cell proliferation and induced cell cycle arrest and apoptosis of pancreatic cancer cells in vitro, and decreased tumor growth in vivo. Specifically, Bcl-2, Claspin, METTL3 and YTHDF3 were identified as the potential targets of celastrol treatment in pancreatic cancer cells. Moreover, our results indicated that celastrol treatment downregulated METTL3 and decreased mA levels of Claspin and Bcl-2 mRNA, leading to the degradation of Claspin and Bcl-2 mRNA in pancreatic cancer cells. Furthermore, we revealed that celastrol treatment downregulated Claspin and Bcl-2, at least in part, in an mA-YTHDF3-mediated manner in pancreatic cancer cells.
Our study highlighted a novel mechanism underlying celastrol-induced cellular proliferation inhibition and apoptosis in pancreatic cancer cells via mA-YTHDF3-mediated downregulation of Claspin and Bcl-2.
已显示雷公藤红素具有抗癌药理活性，然而，雷公藤红素参与胰腺癌的分子机制仍有待进一步阐明。本研究旨在阐明雷公藤红素是否通过调节RNA m⁶A修饰来抑制胰腺癌。
通过CCK-8测定、EdU测定、集落形成测定、流式细胞术分析和皮下异种移植实验评估雷公藤红素处理对胰腺癌细胞恶性表型的影响。进行RNA测序（RNA-seq）分析以分析雷公藤红素处理的胰腺癌细胞中差异表达的基因。采用RT-qPCR、蛋白质免疫印迹和免疫组织化学法评估所示基因的表达。应用RNA斑点杂交和总RNA m⁶A修饰定量测定、MeRIP-qPCR测定、RIP-qPCR测定、RNA稳定性和蛋白质稳定性测定来评估雷公藤红素处理对胰腺癌细胞的调控机制。
我们证明雷公藤红素在体外抑制胰腺癌细胞的增殖并诱导细胞周期停滞和凋亡，在体内减少肿瘤生长。具体而言，Bcl-2、Claspin、METTL3和YTHDF3被确定为雷公藤红素处理胰腺癌细胞的潜在靶点。此外，我们的结果表明，雷公藤红素处理下调METTL3并降低Claspin和Bcl-2 mRNA的m⁶A水平，导致胰腺癌细胞中Claspin和Bcl-2 mRNA的降解。此外，我们发现雷公藤红素处理至少部分地以m⁶A-YTHDF3介导的方式下调胰腺癌细胞中的Claspin和Bcl-2。
我们的研究突出了雷公藤红素通过m⁶A-YTHDF3介导的Claspin和Bcl-2下调诱导胰腺癌细胞增殖抑制和凋亡的新机制。
