Due to LSCC’s uncertain molecular mechanism, over 60% of patients were already at late stages when the disease was discovered ( Steuer et al., 2017). Due to the proneness to recurrence and metastasis, the five-year overall survival rate for LSCC has been approximately 50% in recent years ( Cavaliere et al., 2021). ![]() Research shows that nearly 180,000 new cases of throat cancer and nearly 100,000 throat cancer deaths were reported worldwide in one year ( Bray et al., 2018). 95 to 98% of laryngeal cancers are squamous cell carcinomas (LSCC) ( Zuo et al., 2016). Laryngeal cancer is not only the most common malignancy of the head and neck, but also the second most common respiratory system tumor after lung cancer. Our study further sheds light on the molecular mechanism of LTA4H as a clinical therapy target for LSCC. These results suggest that LTA4H may combine with genes associated with LSCC as an RNA-binding protein to perform a cancer regulatory function. QRT-PCR validation confirmed that LTA4H specifically binds to mRNAs of carcinogenesis-associated genes, including LTBP3, ROR2, EGFR, HSP90B1, and lncRNAs represented by NEAT1. More notably, LTA4H-binding genes were significantly enriched in the mitotic cell cycle, DNA repair, RNA splicing-related pathways, and RNA metabolism pathways, which means that LTA4H has tumor-related alternative splicing regulatory functions. In the LTA4H binding peak, the frequency of the AAGG motif reported to interact with TRA2 β4 was high in both replicates. We found that LTA4H extensively binds with mRNAs/pre-mRNAs and lncRNAs. To investigate the potential role of LTA4H in LSCC, we employed the improved RNA immunoprecipitation and sequencing (iRIP-Seq) experiment to get the expression profile of LTA4H binding RNA in HeLa model cells, a cancer model cell that is frequently used in molecular mechanism research. Recent research shows that the increased expression of LTA4H in laryngeal squamous cell carcinoma (LSCC) promotes tumor proliferation, migration, and metastasis. The RNA-binding metabolic enzyme LTA4H is a novel target for cancer chemoprevention and chemotherapy. (5) Splice reads: statistics of sequencing sequences (also known as Junction reads) that are segmented to two exons (6) Non splice reads: whole-segment comparison of sequencing sequences to exons (7) rmdup reads: remove duplicate reads, remove the PCR duplication reads, that is, the total number of reads counted after all the repeated reads are counted only once, and their proportion in unique mapped reads (8) sense reads: reads on the sense chain (9) antisense reads: reads matched to the antisense chain. (4) Total Multiple mapped: The number of reads with multiple locations on the reference genome and their proportion in total mapped reads. Table S2: Mapping of clean reads on the reference genome (1) Total reads: input data or clean reads, the number of filtered reads after sequencing sequence (2) Total mapped: the number of sequences that can be mapped to the genome (3) Total Uniquely mapped: The number of reads uniquely located on the reference genome and their proportion in total mapped reads. The ratio of duplicate reads to total reads. Table S1: The form shows the quality control of our iRIP-sequence is good (1) Raw data: the number of original sequences transformed from the original image data obtained by sequencing through base calling (2) Clean reads: the raw reads are stripped of the adapter sequence, and the number of valid sequences obtained after low-quality bases is used for subsequent analysis (3) Clean Per: the proportion of clean reads in raw reads (4) Raw base: the count the number of bases it contains, based on the number and length of raw reads, in G (5) Clean base: according to the number and length of clean reads, count the number of bases it contains, in G (6) Base Per: the proportion of Clean base to Raw base (7) Unique tag: unique tag, the number of non-repeating reads and its proportion of clean reads (8) Q20: Proportion of bases whose sequencing error rate is less than 1% (9) Q30: Proportion of bases whose sequencing error rate is less than 0.1% (10) GC: the percentage of GC bases in the total number of bases (11) DUP: duplication level.
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