Abstract: This wok aims to elucidate the regulation of 9R-P201 on hepatoma cells（HepG2）at transcriptome level by investigating the gene fusion，SNP（Single Nucleotide Polymorphism）mutation，alternative splicing and other events after 9R-P201 treating HepG2，and analyzing the biological processes and signaling pathways involved by differentially expressed genes. The expression differences of the genes before and after the 9R-P201 treating HepG2 cell line were detected by transcriptome sequencing. Meanwhile，gene fusion，SNPs，and alternative splicing were identified by tophat-fusion，SAMTOOLS software，and rMATS respectively. Functional enrichment analysis of differentially expressed genes were performed by GO（Gene Ontology）and KEGG（Kyoto Encyclopedia of Genes and Genomes）. As results，276 alternative splicing events，5 557 SNP sites，and 45 gene fusion events were detected in the transcriptome sequencing. In addition，403 differentially expressed genes including 269 up-regulated and 134 down-regulated were detected. Gene GO and KEGG enrichment analysis showed that differentially expressed genes were significantly involved in the biological processes of cell growth，locomotion，as well as a number of cancer-related signaling pathways. In conclusion，the study revealed that the differentially expressed genes resulted from 9R-P201 treating HepG2 were significantly correlated with the biological processes and signaling pathways related to cancer and hepatocellular carcinoma HepG2 cell line，and a large number of alternative splicing events，SNP mutations，gene fusion events after 9R-P201 inducement occurred，suggesting this peptide may be used as a potential drug for subsequent hepatocellular carcinoma interventional therapy.

Key words: 9R-P201, hepatocellular carcinoma, transcriptome sequencing, gene