Identification and functional analysis of genes which confer resistance to cisplatin in tumor cells

• Published in: Biochemical pharmacology 2010-07, Vol.80 (2), p.262-276
• Main Authors: Wu, Zchong-Zcho, Lu, Hsing-Pang, Chao, Chuck C.-K.
• Format: Article
• Language: English
• Subjects: Antineoplastic Agents - pharmacology; Biological and medical sciences; Chemotherapy; Cisplatin - pharmacology; Cisplatin resistance; CITED2; Drug Resistance, Neoplasm - drug effects; Drug Resistance, Neoplasm - genetics; Gene Expression Profiling; Gene Expression Regulation, Neoplastic - drug effects; Gene Expression Regulation, Neoplastic - genetics; Gene Knockdown Techniques; Gene Silencing; HeLa Cells - drug effects; Humans; Inverted Repeat Sequences - genetics; Kidney - drug effects; Kidney - embryology; Medical sciences; NAPA; Oligonucleotide Array Sequence Analysis; Pharmacology. Drug treatments; shRNA
• ISSN: 0006-2952; 1873-2968
• DOI: 10.1016/j.bcp.2010.03.029

The present model illustrates that cisplatin activates the expression of cisplatin resistance (CPR) genes in treated cancer cells. Upregulation of CPR genes may help cancer cells to survive against cisplatin stress, whereas inhibition of CPR gene activity with short-hairpin RNA (shRNA) may lead to apoptosis. The efficacy of cisplatin during cancer chemotherapy is often impaired by the emergence of cancer cells which become resistant to chemotherapeutic agents. While various mechanisms have been proposed to explain resistance to cisplatin, the genes involved in this process still remain unclear. By using DNA microarrays, we performed a genome-wide analysis of cisplatin-resistant HeLa cells in order to identify genes involved in resistance. We identified nine genes ( NAPA, CITED2, CABIN1, ADM, HIST1H1A, EHD1, MARK2, PTPN21, and MVD), which were consistently upregulated in two cisplatin-resistant HeLa cell lines. The upregulated genes, here referred to as cisplatin resistance genes ( CPR), were further analyzed for their ability to modify the response of HEK293 cells to cisplatin. Short-hairpin RNA (shRNA) knockdown of CPR genes, individually or in combination, was shown to sensitize HEK293 cells to cisplatin, but not to vincristine or taxol, suggesting that CPR genes may be involved specifically in cisplatin resistance. Among the treatments performed, shRNA knockdown of NAPA was the most efficient treatment able to sensitize cells to cisplatin. Furthermore, shRNA knockdown of a single CPR gene was sufficient to partially reverse acquired cisplatin resistance in HeLa cells. Sensitization to cisplatin following knockdown of CPR genes was also observed in the tumorigenic cell lines Sk-ov-3, H1155, and CG-1. Based on these results, we propose that the CPR genes identified here may represent potential candidates for novel target therapies aimed at preventing resistance to cisplatin during chemotherapy.