Abstract
Purpose
The expression of eukaryotic translation initiation factor-2 subunit 3 (EIF2S3) in patients with non-small cell lung and colorectal cancer is lower than that in healthy individuals. However, the functions of EIF2S3 remain unclear, and its study in leukemia has not been reported. The article aims to explore the role of EIF2S3 in AML (acute myeloid leukemia) and its underlying mechanism.
Methods
Reverse transcription-quantitative PCR was performed to evaluate the expression levels of EIF2S3, and its association with patient prognosis was determined. Inducible HEL-EIF2S3 and HL-60-EIF2S3 cell lines were established by retrovirus infection. Cellular proliferation and the cell cycle were analyzed using Cell Counting Kit-8 and flow cytometric analyses. Tumorigenic ability was evaluated using xenograft nude mouse model. Gene expression profiles were analyzed in HL-60-EIF2S3 cells by next-generation sequencing, and WB analysis was performed to detect the expression of related proteins.
Results
The expression of EIF2S3 in patients with AML was lower than that experiencing CR (P = 0.02). Furthermore, EIF2S3 overexpression inhibited cellular proliferation, halted G0/1 to S phase cell cycle progression, and inhibited tumorigenicity (P = 0.015). 479 differentially expressed genes were identified between HL60-EIF2S3 DOX (−) and HL60-EIF2S3 DOX ( +) cells via NGS and several of them involved in MAPK/ERK signaling pathway. The phosphorylation levels of ERK decreased when EIF2S3 was overexpressed (P < 0.050).
Conclusion
EIF2S3 overexpression may result in a decrease in cellular proliferation, cell cycle arrest, and tumorigenic inhibition via the MAPK/ERK signaling pathway in AML cells.
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Availability of data and materials
The datasets used and/or analyzed during the current study are available from the corresponding author on reasonable request.
Code availability
Not applicable.
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Funding
The present study was supported by the National Natural Science Foundation of China (Grant no. 81672661, 81870113) and the Guangzhou Science and Technology Project (grant no. 201804010199).
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LX, YZ, and ZH conceived the study idea, supervised the project and designed the experiments; JL and SC performed the experiments and wrote the manuscript; LL, HT, BL, YC, and XZ collected the samples, performed the experiments, and analyzed the data.
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The present study was approved by the ethics committee of the First Affiliated Hospital of Guangzhou Medical University. All patients provided written informed consent for the use of their clinical specimens. Animals were purchased from the Animal experimental center, Shanghai, China, and the protocol was approved by the Institutional Animal Care and Use Committee of Guangzhou Medical University. All mouse procedures were performed in strict accordance with the recommendations of the Guide for the Care and Use of Laboratory Animals of Guangzhou Medical University. Animal experiments were performed at Guangzhou Medical University and all surgeries were performed under sodium pentobarbital anesthesia; all efforts were made to minimize animal suffering.
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432_2021_3712_MOESM1_ESM.tif
Supplementary file1 Fig. S1 EIF2S3 mRNA expression level in AML patients of TCGA based on classification of clinical characteristics. A The difference of EIF2S3 mRNA expression based on FBA types. EIF2S3 expression in M3 was lower than M0 (P=0.019), M1 (P=0.001), M2 (P=0.004) and M4 (P=0.022) patients. EIF2S3 expression in M5 patients was lower than M0 (P=0.012), M1 (P<0.001), M2 (P<0.001) and M4 (P=0.003) patients. There were no significant differences between other types. B The difference of EIF2S3 mRNA expression based on gender (P=0.015). C The difference of EIF2S3 mRNA expression based on age. EIF2S3 expression in 21-40 years-old patient was higher than 81-100 years-old patients (P=0.048). There were no significant differences in other age groups. D The difference of EIF2S3 mRNA expression based on races (P>0.05) (TIF 634 KB)
432_2021_3712_MOESM2_ESM.tif
Supplementary file2 Fig. S2 EIF2S3 mRNA expression level in AML patients of TCGA based on classification of genetic abnormality. A The difference of EIF2S3 mRNA expression based on FLT3 mutation or not (P>0.05). B The difference of EIF2S3 mRNA expression based on PML/RAR fusion or not (P>0.05). C The difference of EIF2S3 mRNA expression based on RAS activation or not (P>0.05) (TIF 14657 KB)
432_2021_3712_MOESM3_ESM.tif
Supplementary file3 Fig. S3 EIF2S3 mRNA expression levels in patients with primary AML (PB) and healthy controls. Comparing with health controls, EIF2S3 expression was downregulated in primary AML patients (PB) (P =0.009). PB, peripheral blood (TIF 81 KB)
432_2021_3712_MOESM4_ESM.tif
Supplementary file4 Fig. S4 Identify the DEGs in HL60-EIF2S3 DOX (-) and HL60-EIF2S3 DOX (+) by NGS. A Using P<0.05 and |log2FC|≥2 as cut-0ff criteria, the volcano plot of DEGs showed there were totally 479 DEGs, including 271 upregulated genes (red dots) and 208 downregulated genes (blue dots). B In the hot plot of top 30 DEGs, red represents upregulation and blue represent downregulation. The closer the color was to the ends of the color column, the greater the difference (TIF 9749 KB)
432_2021_3712_MOESM5_ESM.tif
Supplementary file5 Fig. S5 GO enrichment analysis in HL60-EIF2S3 DOX (-) and HL60-EIF2S3 DOX (+). The top 30 of GO enrichment described relevant biological processes of DEGs. The color of the dots represents the significance of q value, and the size of the dots represents the number of DEGs involved in the GO domain (TIF 24273 KB)
432_2021_3712_MOESM6_ESM.tif
Supplementary file6 Fig. S6 KEGG enrichment analysis in HL60-EIF2S3 DOX (-) and HL60-EIF2S3 DOX (+). The top 30 of pathway enrichment showed that DEGs primarily enriched in MAPK pathway. The color of the dots represents the significance of q value, and the size of the dots represents the number of DEGs involved in the different pathways (TIF 11516 KB)
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Supplementary file7 Fig. S7 RT-qPCR showed the relative mRNA expression level of 13 DEGs in MAPK signaling pathway. *P < 0.05, **P < 0.01, and ***P < 0.001 (TIF 473 KB)
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Lu, J., Chen, S., Tan, H. et al. Eukaryotic initiation factor-2, gamma subunit, suppresses proliferation and regulates the cell cycle via the MAPK/ERK signaling pathway in acute myeloid leukemia. J Cancer Res Clin Oncol 147, 3157–3168 (2021). https://doi.org/10.1007/s00432-021-03712-5
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DOI: https://doi.org/10.1007/s00432-021-03712-5