Establishment and gene analysis of a cisplatin-resistant cell line, Sa-3R, derived from oral squamous cell carcinoma
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Published on: September 10, 2014
Ken Nakatani1, Megumi Nakamura, Katsuhiro Uzawa
1Department of Oral and Maxillofacial Surgery, School of Medicine, Wakayama Medical University, Kimiidera 811-1, Wakayama 641-8509, Japan.
Abstract:
Cisplatin (CDDP) is widely used for chemotherapy of many malignancies, especially of oral squamous cell carcinoma (SCC). However, because the mechanism of resistance to CDDP is unclear, we established a CDDP-resistant cell line, Sa-3R, from a CDDP-sensitive cell line, Sa-3, which was derived from moderately differentiated SCC of the lower gingiva. The 3-(3,4-dimethyl-thiazol-2-yl) 2,5-diphenyltetrazolium bromide assay indicated that Sa-3R has 7.5-fold greater resistance to CDDP than Sa-3. Comparing gene expression levels in the cell lines using an in-house cDNA microarray, which represented 2,201 oral disease origin genes, many differentially expressed genes were identified. The ATP-binding cassette transporter genes (MDR-1, MRP-1, and MRP-2), and FANCONI, GRP58, FLJ12089, and SPINT-2 were up-regulated, whereas FOSL1, MRPS27, and PGK-1 were down-regulated. These results were confirmed by semiquantitative reverse transcriptase-polymerase chain reaction. The Sa-3/Sa-3R cell lines could be useful to identify the candidates responsible for the mechanism of CDDP-resistance and the up- or down-regulated genes identified by the gene expression profiles in the Sa-3R cell line may be, in part, associated with the mechanism.
Insights
Researchers developed a cisplatin-resistant oral cancer cell line (Sa-3R) to study chemotherapy resistance mechanisms. Gene expression analysis revealed key up-regulated and down-regulated genes potentially involved in cisplatin resistance in oral squamous cell carcinoma.
Area of Science:
- Oncology
- Molecular Biology
- Genetics
Background:
- Cisplatin (CDDP) is a cornerstone chemotherapy for various cancers, including oral squamous cell carcinoma (SCC).
- The precise mechanisms underlying CDDP resistance remain incompletely understood, hindering treatment optimization.
- Developing reliable models is crucial for elucidating resistance pathways.
Purpose of the Study:
- To establish and characterize a cisplatin-resistant cell line (Sa-3R) from a cisplatin-sensitive oral SCC cell line (Sa-3).
- To compare gene expression profiles between sensitive and resistant cell lines to identify potential mechanisms of CDDP resistance.
Main Methods:
- Generation of a CDDP-resistant cell line (Sa-3R) from an oral SCC cell line (Sa-3).
- Assessment of CDDP resistance using the 3-(3,4-dimethyl-thiazol-2-yl) 2,5-diphenyltetrazolium bromide (MTT) assay.
- Genome-wide gene expression analysis using an in-house cDNA microarray representing 2,201 oral disease-related genes.
- Validation of differentially expressed genes using semiquantitative reverse transcriptase-polymerase chain reaction (RT-PCR).
Main Results:
- The Sa-3R cell line exhibited 7.5-fold greater resistance to CDDP compared to the parental Sa-3 cell line.
- Microarray analysis identified significant differences in gene expression, with notable up-regulation of ATP-binding cassette transporter genes (MDR-1, MRP-1, MRP-2), FANCONI, GRP58, FLJ12089, and SPINT-2.
- Down-regulation was observed for FOSL1, MRPS27, and PGK-1 in the resistant cell line.
- RT-PCR confirmed the differential expression patterns observed in the microarray analysis.
Conclusions:
- The established Sa-3/Sa-3R cell line pair serves as a valuable tool for investigating CDDP resistance mechanisms in oral SCC.
- The identified up-regulated and down-regulated genes are potentially implicated in the development of cisplatin resistance.
- Further studies utilizing this model can help pinpoint specific molecular targets for overcoming chemotherapy resistance.
