Ibuprofen promotes p75 neurotrophin receptor expression through modifying promoter methylation and N6-methyladenosine-RNA-methylation in human gastric cancer cells

ABSTRACT

It is acknowledged that nonsteroidal anti-inflammatory drugs (NSAIDs) can participate in various signaling pathways, while information about their epigenetic effects are limited. p75NTR (p75 neurotrophin receptor) can inhibit tumor growth by inducing cell cycle arrest and regulating cell cycle arrest and apoptotic cell death. The expression of p75NTR is influenced by epigenetic roles. We explored the effects of ibuprofen on p75NTR expression and investigated whether promoter methylation and N6-methyladenosine (m6A) RNA methylation regulates this process in human gastric cancer cells (SGC7901 and MKN45). Cell lines were treated with ibuprofen 0, 2.5, 5, 10, 20 µM, and then DNA, RNA, and protein were isolated 24 h later. Expression and promoter methylation of p75NTR were detected by RT-qPCR and Western blot. The levels of m6A-p75NTR were measured by RNA immunoprecipitation. We also used RT-qPCR to determine the levels of m6A-related regulators, METTL3, METTL14, ALKBH5, FTO, YTHDC2, and YTHDF1-3. Ibuprofen attenuated p75NTR promoter methylation (p < 0.01) and increased p75NTR level (p < 0.001). Ibuprofen increased m6A-p53 expression (p < 0.01) by promoting the expression of METTL3 (p < 0.01) and METTL14 (p < 0.05); and increased levels of YTHDF1 (p < 0.001), YTHDF3 (p < 0.001), and YTHDC2 (p < 0.01) that finally reinforced p53 translation (p < 0.01). Therefore, our results present that ibuprofen epigenetically increased p75NTR expression by downregulating promoter methylation and upregulating m6A-RNA-methylation in SGC7901 and MKN45 cells. Our study unveils a novel mechanism for p75NTR regulation by NSAIDs and helps the design of treatment targets.

GRAPHICAL ABSTRACT

KEYWORDS:

• Ibuprofen
• promoter methylation
• m6A-RNA-methylation
• p75NTR
• gastric cancer

Disclosure statement

No potential conflict of interest was reported by the author(s).

Author contributions

Conception and design: Haifeng Jin; Acquisition of data: Haifeng Jin, Zheng Wu, Bibo Tan, Zhen Liu, Zhanfei Zu; Analysis and interpretation of data: Zhanfei Zu, Xiaoyun Wu, Yuwang Bi, Xingmao Hu; Writing, review, and/or revision of the manuscript: Haifeng Jin, Zheng Wu, Bibo Tan; Study supervision: Haifeng Jin.

Statement of Ethics

This study was approved by the ethics of the Ethics Committee of The 980th Hospital of the PLA Joint Logistics Support Force (No. 202109_247).

Competing statement

The authors declare no conflict of interest.

Data availability statement

All data and information can be obtained from the corresponding author on reasonable request.

Supplementary material

Supplemental data for this article can be accessed online at https://doi.org/10.1080/21655979.2022.2092674

Additional information

Funding

This work was funded by the Key science and technology research project of Hebei Provincial Health Commission [20210313]; Self-funded project of Shijiazhuang Science and Technology Bureau [201200783]; and Youth Science and Technology Project of Hebei Health Commission [20191219].