Proteomic analysis of proteins responsible for cisplatin-resistant nasopharyngeal carcinoma cell lines

Name: Parisa Prathaphan

LCSH: Kasetsart University -- Theses. M.S. (Biochemistry) 2021

Classification :.LCCS: RC280.P4

LCSH: Kasetsart University. -- Department of Biochemistry -- Theses

LCSH: Nasopharyngeal carcinoma

LCSH: Nasopharynx -- Cancer -- Chemotherapy

LCSH: Nasopharynx -- Tumors

LCSH: Proteomics

LCSH: Cisplatin

LCSH: Drug resistance in cancer cells

Abstract: Nasopharyngeal carcinoma (NPC) is a cancer of a nasopharynx. A major problem of the treatment is a chemoresistance. As a result, mechanisms involved in the drug resistance are needed to be investigated for finding efficient treatment. Here, the aims of this study are to establish cisplatin-resistant NPC cell lines and investigate their protein expression profiles to reveal the chemoresistant mechanisms. First, NPC cell lines (5-8F and 6-10B) were continuously treated with increasing concentrations of cisplatin to develop cisplatin-resistant 5-8F and 6-10B (5-8FcisR and 6-10BcisR) cells. The half-maximal inhibitory concentration (IC50) value of the 5-8FcisR and 6- 10BcisR cells significantly increased with resistant index (RI) of 8.3 ± 0.9 and 2.4 ± 0.2, respectively. Additionally, the cisplatin-resistant NPC cell lines had slower growth rates than their parental NPC cell lines. The doubling times of 5-8FcisR and 6- 10BcisR were significantly increased from 19.7 ± 0.8 to 22.7 ± 0.9 hr in 5-8FcisR and 22 ± 0.5 to 25.6 ± 1.4 hr in 6-10BcisR. However, the morphologies of 5-8FcisR and 6- 10BcisR cells were approximately similar to those of the parental cells. Besides, a GeLc-MS/MS was performed to identify and quantify proteins for a comparative proteomic analysis. Total 915 proteins were found in 5-8F and 5-8FcisR and 617 proteins were found in 6-10B and 6-10BcisR cells. Altered protein expressions were significantly considered at p < 0.05. Protein interaction and functional enrichment of altered protein expressions were analyzed using String software. The altered proteins in 5-8FcisR were related to ‘programmed cell death’ and ‘cell differentiation’ processes. The altered proteins in 6-10BcisR were related to ‘mRNA metabolic process’, ‘epithelial cell differentiation’, ‘transport’, ‘drug metabolic process’, ‘daunorubicin metabolic process’, and ‘doxorubicin metabolic process’. Twenty-five differentially expressed proteins (i.e. 1433Z, ATPB, CLIC1, DESM, ENO1, ESPL1, FUBP1, FUBP2, GAPDH, GSTP, HMGB1, IMB1, K1C10, KCRB, MDHM, PCNA, PDIA4, PEBP1, PLEC1, PPBI, PRDX2, SFRS3, STMN1, TPM3, and VINC) were validated for their gene expression levels using qRT-PCR. Thirteen out of twentyfive target genes demonstrated a positive correlation between gene transcription and protein expression. In addition, FUBP1 and KCRB were chosen to confirm their differential expressions using western-blot. The result revealed that both proteins also exhibited the corresponding results with the proteomic report.

Kasetsart University. Office of the University Library

Address: Bangkok

Email: tdckulib@ku.ac.th

Name: Pichamon Kiatwuthinon

Role: Thesis Advisor

Name: Napachanok Mongkoldhumrongkul

Role: Thesis CO-Advisor

Name: Onrapak Reamtong

Role: Thesis CO-Advisor

Created: 2021

Modified: 2025-07-20

Issued: 2025-07-20

วิทยานิพนธ์/Thesis

application/pdf

URL: https://www.lib.ku.ac.th/KUthesis/2564/parisa-pra-all.pdf

CallNumber: RC280.P4 .P37

eng

DegreeName: Master of Science

Level: Master's Degree

Descipline: Biochemistry

Grantor: Kasetsart University

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