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Comparative Proteomic Profiling of Cisplatin-resistant Nasopharyngeal Carcinoma Cell Lines: Novel Biomarkers for Improving Chemotherapy of NPC.
Anticancer Research 2022 July
BACKGROUND/AIM: Nasopharyngeal carcinoma (NPC) originates in the hidden nasopharynx, causing NPC patients to be diagnosed at a late stage and develop drug resistance. Therefore, the identification of drug-resistance biomarkers is indispensable to improve NPC detection and treatment. Hence, this study aimed to identify novel cisplatinresistance biomarkers using comparative proteomic profiles of cisplatin-resistant (CDDP/NPC) cell lines.
MATERIALS AND METHODS: Two cisplatin-resistant NPC cell lines (CDDP/5-8F and CDDP/6-10B) were established by a continuous cisplatin treatment. Then, morphology, proliferation, and migration of all NPC cells were evaluated, followed by the examination of protein profiles using 1D in-gel digestion coupled with mass spectrometry. The potential drugresistance biomarkers were transcriptionally and translationally validated by qPCR and western blotting, respectively.
RESULTS: CDDP/5-8F and CDDP/6-10B cells were successfully developed with a resistance index of 8.42 and 2.46, respectively. Furthermore, both CDDP/NPC cells demonstrated relatively altered morphology, retarded growth, and decreased migration. Additionally, the comparative proteomic analysis of CDDP/NPC revealed 92 differentially expressed proteins (DEPs). Specifically, up-regulated DEPs were notably enriched in cellular metabolic processes, while down-regulated DEPs were predominantly enriched in actin filament-based movement, methylation, and programmed cell death. Six up-regulated, namely ALPI, CKB, HMGB1, KHSRP, PDIA4, and STMN1, and three down-regulated proteins, FUBP1, YWHAZ, and PLEC, were validated at the transcriptional level. CKB and FUBP1 were further validated at the translational level and demonstrated corresponding expression levels at both protein and gene levels.
CONCLUSION: Our findings suggest novel biomarkers to indicate cisplatin resistance in NPC, expanding the drug resistance knowledge and paving the way for in-depth mechanism studies in NPC.
MATERIALS AND METHODS: Two cisplatin-resistant NPC cell lines (CDDP/5-8F and CDDP/6-10B) were established by a continuous cisplatin treatment. Then, morphology, proliferation, and migration of all NPC cells were evaluated, followed by the examination of protein profiles using 1D in-gel digestion coupled with mass spectrometry. The potential drugresistance biomarkers were transcriptionally and translationally validated by qPCR and western blotting, respectively.
RESULTS: CDDP/5-8F and CDDP/6-10B cells were successfully developed with a resistance index of 8.42 and 2.46, respectively. Furthermore, both CDDP/NPC cells demonstrated relatively altered morphology, retarded growth, and decreased migration. Additionally, the comparative proteomic analysis of CDDP/NPC revealed 92 differentially expressed proteins (DEPs). Specifically, up-regulated DEPs were notably enriched in cellular metabolic processes, while down-regulated DEPs were predominantly enriched in actin filament-based movement, methylation, and programmed cell death. Six up-regulated, namely ALPI, CKB, HMGB1, KHSRP, PDIA4, and STMN1, and three down-regulated proteins, FUBP1, YWHAZ, and PLEC, were validated at the transcriptional level. CKB and FUBP1 were further validated at the translational level and demonstrated corresponding expression levels at both protein and gene levels.
CONCLUSION: Our findings suggest novel biomarkers to indicate cisplatin resistance in NPC, expanding the drug resistance knowledge and paving the way for in-depth mechanism studies in NPC.
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