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02 April 2019 : Laboratory Research  

Next-Generation Sequencing Analysis of mRNA Profile in Cisplatin-Resistant Gastric Cancer Cell Line SGC7901

Zhenwei Deng1AB, Huaiming Wang2AC, Guohu Guo3CDF, Xiyao Li14DE, Yongchang Cai1BD, Yuxin Tang1DE, Yijun Wang1CF, Jiabao Li1BD, Zhibin Lu1BF, Xueqiao Yu5ABC, Ruiping Li1DE, Libo Li1ABF*

DOI: 10.12659/MSM.915866

Med Sci Monit 2019; 25:2386-2396

Abstract

BACKGROUND: Cisplatin-resistant gastric cancer (GC) occurs in patients with GC treated with cisplatin-based chemotherapy, which results in disease progression and early recurrence during the treatment.

MATERIAL AND METHODS: To understand the initiation and developmental mechanism underlying cisplatin-resistant GC, we developed cisplatin-resistant SGC7901 cells (SGC7901/DDP) from the parental cells (SGC7901/S) by continuous exposure to increasing concentrations of cisplatin and subjected these 2 cell lines to RNA sequencing analysis. The data were verified by quantitative polymerase chain reaction and their functional role was evaluated by cell counting kit 8 assay and cell apoptosis and cell cycle flow cytometric analysis. Bioinformatics analysis was performed to classify the differentially-expressed genes (DEGs) involved in the development of cisplatin resistance.

RESULTS: In comparison with SGC7901/S cells, SGC7901/DDP cells showed a total of 3165 DEGs (2014 upregulated and 1151 downregulated, fold change ≥2, and adjusted P value <0.001). qRT-PCR confirmed the reliability of the RNA sequencing results. Depletion of the top 5 upregulated mRNAs reversed the resistant index, increased apoptotic SGC7901/DDP cells, and arrested the cells at G2/M phase. Gene ontology analysis revealed that the DEGs mainly regulate metabolic process, immune system, locomotion, cell adhesion, cell growth, cell death, cytoskeleton organization, cell binding, signal transducing activity, and antioxidant activity. Kyoto Encyclopedia of Genes and Genomes analysis showed that the DEGs were mainly involved in the PI3K-Akt signaling pathway, Rap1 signaling pathway, proteoglycans in cancer, regulation of actin cytoskeleton, and pathways in cancer.

CONCLUSIONS: The present study is the first to interrogate mRNAs profiles in human GC cells with cisplatin resistance using RNA sequencing, which may assist in discovering potential therapeutic targets for cisplatin-resistant GC patients.

Keywords: Cisplatin, Drug Resistance, Sequence Analysis, RNA, Stomach Neoplasms, Apoptosis, Drug Resistance, Multiple, Drug Resistance, Neoplasm, Gene Expression Profiling, Gene Expression Regulation, Neoplastic, High-Throughput Nucleotide Sequencing, RNA, Messenger, Reproducibility of Results, transcriptome

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DOI: 10.12659/MSM.938365

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750