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Phenethyl isothiocyanate inhibits metastasis potential of non-small cell lung cancer cells through FTO mediated TLE1 m6A modification

Acta Pharmacologica Sinica volume 45, pages 619–632 (2024)Cite this article

Abstract

N6-methyladenosine (m6A) modification is a prevalent RNA epigenetic modification, which plays a crucial role in tumor progression including metastasis. Isothiocyanates (ITCs) are natural compounds and inhibit the tumorigenesis of various cancers. Our previous studies show that ITCs inhibit the proliferation and metastasis of non-small cell lung cancer (NSCLC) cells, and have synergistic effects with chemotherapy drugs. In this study, we investigated the molecular mechanisms underlying the inhibitory effects of ITCs on cancer cell metastasis. We showed that phenethyl isothiocyanate (PEITC) dose-dependently inhibited the cell viability of both NSCLC cell lines H1299 and H226 with IC50 values of 17.6 and 15.2 μM, respectively. Furthermore, PEITC dose-dependently inhibited the invasion and migration of H1299 and H226 cells. We demonstrated that PEITC treatment dose-dependently increased m6A methylation levels and inhibited the expression of the m6A demethylase fat mass and obesity-associated protein (FTO) in H1299 and H226 cells. Knockdown of FTO significantly increased m6A methylation in H1299 and H226 cells, impaired their abilities of invasion and migration in vitro, and enhanced the inhibition of PEITC on tumor growth in vivo. Overexpression of FTO promoted the migration of NSCLC cells, and also mitigated the inhibitory effect of PEITC on migration of NSCLC cells. Furthermore, we found that FTO regulated the mRNA m6A modification of a transcriptional co-repressor Transducin-Like Enhancer of split-1 (TLE1) and further affected its stability and expression. TCGA database analysis revealed TLE1 was upregulated in NSCLC tissues compared to normal tissues, which might be correlated with the metastasis status. Moreover, we showed that PEITC suppressed the migration of NSCLC cells by inhibiting TLE1 expression and downstream Akt/NF-κB pathway. This study reveals a novel mechanism underlying ITC’s inhibitory effect on metastasis of lung cancer cells, and provided valuable information for developing new therapeutics for lung cancer by targeting m6A methylation.

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Fig. 1: PEITC inhibited the growth, invasion and migration of NSCLC cells.
Fig. 2: PEITC elevated the RNA m6A modification level and inhibited the expression of FTO in NSCLC cells.
Fig. 3: FTO mediated the effect of PEITC on the m6A modifications, invasion and migration of NSCLC cells.
Fig. 4: PEITC regulated m6A modification and expression of genes.
Fig. 5: PEITC regulated m6A modification and expression of TLE1.
Fig. 6: PEITC inhibited the migration of NSCLC cells via TLE1 mediated Akt/NF-κB pathway.
Fig. 7: FTO knockdown enhanced the inhibitory effect of PEITC on NSCLC cell growth in vivo.
Fig. 8: A proposed model of PEITC’s regulation on m6A modification in NSCLC cells.

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Acknowledgements

This work was supported by grants from the National Natural Science Foundation of China (81372519), the Natural Science Foundation of Tianjin (22JCZDJC00450, 21JCQNJC00150), the Project of Health Commission of Tianjin (TJWJ2021MS006), the Tianjin Key Medical Discipline (Specialty) Construction Project (TJYXZDXK-061B), the Project of Tianjin Municipal Education Commission (2020KJ150, 2021KJ212) and the Open Fund of Ministry of Education Key Laboratory of Molecular Microbiology and Technology, Nankai University (NKU-KLMMTME-KFKT-202101).

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  1. These authors contributed equally: Qi-cheng Zhang, Yong-mei Qian, Ying-hui Ren

Authors and Affiliations

  1. Tianjin Key Laboratory of Lung Cancer Metastasis and Tumor Microenvironment, Tianjin Lung Cancer Institute, Tianjin Medical University General Hospital, Tianjin, 300052, China

    Qi-cheng Zhang, Yong-mei Qian, Li-min Cao, Si-jia Zheng, Bing-bing Li, Min Wang & Ke Xu

  2. Department of Anesthesiology, Tianjin First Central Hospital, Tianjin, 300192, China

    Ying-hui Ren

  3. Department of Clinical Laboratory, Shandong Cancer Hospital and Institute, Shandong First Medical University and Shandong Academy of Medical Sciences, Jinan, 250117, China

    Meng-meng Chen

  4. Core Facility Center, Tianjin Medical University General Hospital, Tianjin, 300052, China

    Xiang Wu

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Contributions

KX, QCZ, LMC and YHR provided grant support. QCZ, YMQ, YHR, MMC, LMC, SJZ and BBL performed the experiments. MW and XW performed data analysis. KX and QCZ contributed to the study design and wrote the manuscript. All authors have read and approved the final manuscript.

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Correspondence to Ke Xu.

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Zhang, Qc., Qian, Ym., Ren, Yh. et al. Phenethyl isothiocyanate inhibits metastasis potential of non-small cell lung cancer cells through FTO mediated TLE1 m6A modification. Acta Pharmacol Sin 45, 619–632 (2024). https://doi.org/10.1038/s41401-023-01178-4

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