Abstract
Background
Nasopharyngeal carcinoma (NPC) has high incidence in Southern China and is derived from the mucosal epithelium of the nasopharynx. Accumulating evidence has revealed that peptidyl arginine deiminase 4 (PAD4) exerts carcinogenic effect on certain cancers. We designed this study to probe the specific role that PAD4 plays in NPC and its molecular mechanism.
Methods
PAD4 expression in NPC cells was detected by RT-qPCR analysis. MTT, colony formation, flow cytometry, TUNEL staining, and LC3-II punctuation experiments were done to probe into the biological functions of PAD4 on NPC cellular behaviors in vitro. Subsequently, the upstream regulatory mechanism of PAD4 was investigated by luciferase reporter, RNA pull-down, and RIP assays. The impact of PAD4 on NPC tumor growth in mice was assessed by in vivo xenograft tumor assay.
Results
PAD4 was upregulated in NPC cells. PAD4 knockdown suppressed proliferative ability and promoted apoptosis and autophagy in NPC cells. Additionally, PAD4 expression was negatively regulated by microRNA 3164 (miR-3164). LINC00324 positively upregulated PAD4 expression by interacting with miR-3164 and recruiting HuR protein. The LINC00324/miR-3164/PAD4 axis modulated the PI3K/AKT pathway in NPC cells. Moreover, PAD4 upregulation countervailed the influences of LINC00324 deficiency on NPC cell proliferation, apoptosis, and autophagy and on NPC tumor growth in mice.
Conclusion
LINC00324 promoted NPC malignancy by upregulation of PAD4 to activate the PI3K/AKT pathway.
Graphical abstract
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Data availability
The datasets used or analyzed during the current study are available from the corresponding author on reasonable request.
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Acknowledgements
Thank you for all lab members involved in this study.
Funding
The work was supported by Guangxi Key Research Project (grant number: AB17195005) and Nanning Excellent Young Scientist Program and Guangxi Beibu Gulf Economic Zone Major Talent Program (grant number: RC20190104).
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Hao Chen, Lining Wei, Heming Lu, and Yahua Zhong conceived and designed research; Hao Chen, Lining Wei, Min Luo, Xiaochen Wang, Chaohua Zhu, Huixian Huang, and Xu Liu performed the research; Hao Chen, Lining Wei, Heming Lu, and Yahua Zhong analyzed the data; Hao Chen and Lining Wei wrote the paper; Heming Lu, and Yahua Zhong edited the manuscript. All authors approved final version of manuscript.
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Written informed consents were obtained from all participants. The study complied with the Declaration of Helsinki and was approved by the Ethics Committee of Ethics Committee of People’s Hospital of Guangxi Zhuang Autonomous Region.
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Supplementary Figure 1.
(A) Expression of LINC00324 in NPC tumors and adjacent non-tumor tissues was testified with RT-qPCR. (B) MTT assay evaluated viability of 5-8F and 6-10B cells under indicated transfection. (C) Effects of silenced HuR on expression of LINC00324 or miR-3164 was detected by RT-qPCR. (D) Effects of miR-3164 mimics on expression of LINC00324 or HuR was detected by RT-qPCR. *p < 0.05, **p < 0.01. (PNG 87971 kb)
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Chen, H., Wei, L., Luo, M. et al. LINC00324 suppresses apoptosis and autophagy in nasopharyngeal carcinoma through upregulation of PAD4 and activation of the PI3K/AKT signaling pathway. Cell Biol Toxicol 38, 995–1011 (2022). https://doi.org/10.1007/s10565-021-09632-x
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DOI: https://doi.org/10.1007/s10565-021-09632-x