METTL3 suppresses anlotinib sensitivity by regulating m6A modification of FGFR3 in oral squamous cell carcinoma

Jie Chen; Shuai Li; Zhexun Huang; Congyuan Cao; Anxun Wang; Qianting He

doi:10.1186/s12935-022-02715-7

METTL3 suppresses anlotinib sensitivity by regulating m⁶A modification of FGFR3 in oral squamous cell carcinoma

Cancer Cell Int. 2022 Sep 27;22(1):295. doi: 10.1186/s12935-022-02715-7.

Authors

Jie Chen^#^{1

2}, Shuai Li^#^{1

3}, Zhexun Huang^#⁴, Congyuan Cao¹, Anxun Wang⁵, Qianting He⁶

Affiliations

¹ Department of Oral and Maxillofacial Surgery, The First Affiliated Hospital, Sun Yat-Sen University, Guangzhou, 510080, China.
² Hospital of Stomatology, Guangdong Provincial Key Laboratory of Stomatology, Guanghua School of Stomatology, Sun Yat-Sen University, Guangzhou, 510055, China.
³ Department of Oral and Maxillofacial Surgery, College of Stomatology, Guangxi Medical University, Nanning, China.
⁴ Center of Oral Implantology, Stomatological Hospital, Southern Medical University, Guangzhou, 510280, China.
⁵ Department of Oral and Maxillofacial Surgery, The First Affiliated Hospital, Sun Yat-Sen University, Guangzhou, 510080, China. wanganx@mail.sysu.edu.cn.
⁶ Department of Oral and Maxillofacial Surgery, The First Affiliated Hospital, Sun Yat-Sen University, Guangzhou, 510080, China. heqt3@mail.sysu.edu.cn.

^# Contributed equally.

Abstract

Background: N6-methyladenosine (m⁶A) is an abundant nucleotide modification in mRNA, but there were few studies on its role in cancer drug sensitivity and resistance. Anlotinib has been proved to have effective antitumor effects in oral squamous cell carcinoma (OSCC) in our previous study. Here, we sought to investigate the treatment target of anlotinib and the function and mechanisms of m⁶A modification in regulating anlotinib effect in OSCC.

Methods: Anlotinib treatment in a dose-dependent manner, western blotting, qRT-PCR and cell lost-of-function assays were used to study the treatment target of anlotinib in OSCC. RNA m⁶A dot blot assays, the m⁶A MeRIP-seq and MeRIP-qPCR, RNA and protein stability assays were used to explore the m⁶A modification of the treatment target of anlotinib. Cell lost-of-function assays after METTL3 depletion were conducted to investigate the effect of m⁶A modification level on the therapeutic effect of anlotinib in OSCC. Patient-derived tumor xenograft (PDX) models and immunohistochemistry staining were performed to study the relationship of METTL3 and antitumor sensitivity of anlotinib in vivo.

Results: Anlotinib targeted FGFR3 in the treatment of OSCC and inhibited tumor cell proliferation and promoted apoptosis by inactivating the FGFR3/AKT/mTOR signaling pathway. METTL3 was identified to target and modify FGFR3 m⁶A methylation and then decrease the stability of mRNA. METTL3 expression level was related to the anlotinib sensitivity in OSCC cells in vitro and METTL3 knockdown promoted anlotinib sensitivity of OSCC cells by inhibiting the FGFR3 expression. PDX models samples furthermore showed that METTL3 and FGFR3 levels were tightly correlated with the anlotinib efficacy in OSCC.

Conclusions: In summary, our work revealed that FGFR3 was served as the treatment target of anlotinib and METTL3-mediated FGFR3 m⁶A modification played a critical function in the anlotinib sensitivity in OSCC.

Keywords: Anlotinib; FGFR3; METTL3; OSCC; TKI; m6A methylation.

Abstract

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