Abstract
N6-methyladenosine (m6A) RNA methylation has recently been found involving in regulatory mechanism of the tumor progression. Our aim was to explore the biological function and clinical significance of the m6A methyltransferase METTL3 in intrahepatic cholangiocarcinoma (ICC). In this study, we revealed that METTL3 was upregulated and predicted poor prognosis of patients with ICC. Multivariate regression analysis demonstrated that METTL3 expression was an independent predictor for overall survival in patients with ICC. Moreover, METTL3 knockdown inhibited ICC progression, while METTL3 overexpression showed the opposite effect. METTL3 inhibitor STM2457 also showed anti-tumor effect in ICC. Mechanistically, METTL3 transcription was driven by H3K4me3 activation. Upregulation of METTL3 mediated m6A modification of IFIT2 mRNA and accelerated IFIT2 mRNA decay in a YTHDF2-dependent manner, which promoted the development of ICC and lead to poorer prognosis. In summary, our findings revealed that H3K4me3 activation-driven METTL3 transcription promotes ICC progression by YTHDF2-mediated IFIT2 mRNA degradation, suggesting that METTL3 may serve as a potential target for human ICC therapy.
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Introduction
Intrahepatic cholangiocarcinoma (ICC) is the second most common primary malignant liver cancer, which accounts for ~10% of all such cancers [1, 2]. The prognosis of patients with ICC is poor. Nearly 70% of patients with ICC are already unresectable at the time of diagnosis. Even in patients undergoing curative surgical treatment, the 5-year overall survival (OS) rate is ~30%, and the 5-year recurrence rate is up to 70% [3]. Therefore, effective systemic therapy during the course of the disease is required for the ICC. The combination of gemcitabine and cisplatin is the current first-line therapy for patients with unresectable ICC, but its efficacy remains very limited [4, 31].
RIP-seq or RIP-qPCR
HuCC-T1 and HCCC-9810 Cells were seeded in a 150-mm dish at a density of 1 × 106 cells/ml. The cells were harvested in lysis buffer. The anti-YTHDF2 antibody was used for immunoprecipitated in a dilution of 1:100. The RNA of input and immunoprecipitated samples were isolated with the TRIzol reagent and subjected to sequencing using Illumina HiSeq 2500 platform supplied by Novogene or qPCR analysis. For qPCR analysis, a flank region with no signal was used as a negative control.
Statistical analysis
The statistical analysis of the data was carried out in SPSS 22.0 software and GraphPad Prism 9 software. The data were expressed as mean ± standard deviation and compared by t test, Wilcoxon test, or Chi square test. DFS and OS were measured by Kaplan–Meier method, and the differences between groups were evaluated by log rank test. The independent predictive factors were determined by the cox proportional hazard model. All the statistical analyses, p < 0.05 were considered statistically significant.
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Acknowledgements
The authors greatly acknowledge the financial support from National Natural Science Foundation of China (81772522, 82002501, 82072644, 81972651, and 31771630), the Guangdong Basic and Applied Basic Research Foundation (2021A1515010123, 2019A1515010096 and 2019A1515010686).
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X-YY conceived the study. X-YY, Q-CX and Y-CT designed the study. Q-CX, Y-CT, Y-HS, SC and Y-QZ performed experiments and analyzed the results. Y-HS, C-SH and X-TH provided and assisted in the collection and analysis of clinical samples. X-YY and WZ supervised and guaranteed the study. X-YY, Q-CX, Y-CT and WZ wrote the manuscript. All authors discussed the results and commented on the manuscript, and approved the final version of the manuscript.
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Xu, QC., Tien, YC., Shi, YH. et al. METTL3 promotes intrahepatic cholangiocarcinoma progression by regulating IFIT2 expression in an m6A-YTHDF2-dependent manner. Oncogene 41, 1622–1633 (2022). https://doi.org/10.1038/s41388-022-02185-1
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DOI: https://doi.org/10.1038/s41388-022-02185-1
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