Schenk P W, Boersma A W, Brandsma J A, den Dulk H, Burger H, Stoter G, Brouwer J, Nooter K
Department of Medical Oncology, University Hospital Rotterdam-Daniel den Hoed Cancer Center, Josephine Nefkens Institute, 3000 DR Rotterdam, the Netherlands.
Cancer Res. 2001 Oct 1;61(19):6982-6.
The therapeutic potential of cisplatin, one of the most active and widely used anticancer drugs, is severely limited by the occurrence of cellular resistance. In this study, using budding yeast Saccharomyces cerevisiae as a model organism to identify novel drug resistance genes, we found that disruption of the yeast gene SKY1 (serine/arginine-rich protein-specific kinase from budding yeast) by either transposon insertion or one-step gene replacement conferred cellular resistance to cisplatin. Heterologous expression of the human SKY1 homologue SRPK1 (serine/arginine-rich protein-specific kinase) in SKY1 deletion mutant yeast cells restored cisplatin sensitivity, suggesting that SRPK1 is a cisplatin sensitivity gene, the inactivation of which could lead to cisplatin resistance. Subsequently, we investigated the role of SRPK1 in cisplatin sensitivity and resistance in human ovarian carcinoma A2780 cells using antisense oligodeoxynucleotides. Treatment of A2780 cells with antisense oligodeoxynucleotides directed against the translation initiation site of SRPK1 led to down-regulation of SRPK1 protein and conferred a 4-fold resistance to cisplatin. The human SRPK1 gene has not been associated with drug resistance before. Our new findings strongly suggest that SRPK1 is involved in cisplatin-induced cell kill and indicate that SRPK1 might potentially be of importance for studying clinical drug resistance.
顺铂是最有效的且广泛应用的抗癌药物之一,但其治疗潜力因细胞耐药性的出现而受到严重限制。在本研究中,我们以芽殖酵母酿酒酵母作为模式生物来鉴定新的耐药基因,发现通过转座子插入或一步基因替换破坏酵母基因SKY1(来自芽殖酵母的富含丝氨酸/精氨酸蛋白特异性激酶)可使细胞对顺铂产生耐药性。在SKY1缺失突变酵母细胞中异源表达人SKY1同源物SRPK1(富含丝氨酸/精氨酸蛋白特异性激酶)可恢复顺铂敏感性,这表明SRPK1是一个顺铂敏感性基因,其失活可导致顺铂耐药。随后,我们使用反义寡脱氧核苷酸研究了SRPK1在人卵巢癌A2780细胞顺铂敏感性和耐药性中的作用。用针对SRPK1翻译起始位点的反义寡脱氧核苷酸处理A2780细胞导致SRPK1蛋白下调,并赋予对顺铂4倍的耐药性。此前人类SRPK1基因与耐药性并无关联。我们的新发现强烈表明SRPK1参与顺铂诱导的细胞杀伤,并表明SRPK1可能对研究临床耐药性具有潜在重要性。