Lentiviral short hairpin RNA screen of genes associated with multidrug resistance identifies PRP-4 as a new regulator of chemoresistance in human ovarian cancer.
Lentiviral short hairpin RNA screen of genes associated with multidrug resistance identifies PRP-4 as a new regulator of chemoresistance in human ovarian cancer.
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DOI：
10.1158/1535-7163.mct-08-0316
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发表时间：
2008-08
影响因子：
5.7
通讯作者：
Hornicek, Francis J.
中科院分区：
文献类型：
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作者：
Duan, Zhenfeng;Weinstein, Edward J.;Ji, Diana;Ames, Rachel Y.;Choy, Edwin;Mankin, Henry;Hornicek, Francis J.
已发表的报告暗示了多种可能导致卵巢癌耐药的机制。本研究的主要目的是了解卵巢癌耐药相关基因的过度表达与多药耐药之间的关系。使用慢病毒短发夹 RNA (shRNA) 集合，针对从耐药癌细胞系的转录分析中鉴定出的 132 个基因，在亚致死剂量的紫杉醇存在下进行了个体敲低实验。发现敲除与细胞毒性相关的特定基因包括 MDR1 (ABCB1)、生存素和 PRP-4（前 mRNA 加工因子 4）。这些基因在受到抑制时可以逆转多重耐药细胞系 SKOV-3TR 和 OVCAR8TR 中的紫杉醇耐药性。此前已有报道称，MDR1 和 survivin 在多药耐药和化疗诱导的细胞凋亡中发挥作用；然而，目前尚未认识到 PRP-4 表达对药物敏感性的影响。 PRP-4 属于 Ser/Thr 蛋白激酶家族，在前 mRNA 剪接和细胞有丝分裂中发挥作用，并与 CLK1 相互作用。 Northern 分析表明 PRP-4 在几种紫杉醇耐药细胞系中过表达，并证实 PRP-4 慢病毒 shRNA 可以显着抑制 PRP-4 表达。克隆形成和 MTT 测定均证实 PRP-4 的转录抑制可以将 SKOV-3TR 中的紫杉醇耐药性逆转 5-10 倍。最后，药物敏感细胞中 PRP-4 的过度表达可诱导对紫杉醇、阿霉素和长春新碱的适度耐药性。
Published reports implicate a variety of mechanisms that may contribute to drug resistance in ovarian cancer. The chief aim of this study is to understand the relationship between overexpression of drug resistance associated genes and multidrug resistance in ovarian cancer. Using lentiviral short hairpin RNA (shRNA) collections targeting 132 genes identified from transcriptional profiling of drug resistant cancer cell lines, individual knockdown experiments were performed in the presence of sublethal doses of paclitaxel. Specific genes whose knockdown was found to be associated with cellular toxicity included MDR1 (ABCB1), survivin and PRP-4 (Pre-mRNA Processing factor-4). These genes, when repressed, can reverse paclitaxel resistance in the multidrug resistant cell line SKOV-3TR and OVCAR8TR. Both MDR1 and survivin have been previously reported to play a role in multidrug resistance and chemotherapy induced apoptosis; however, the effect of PRP-4 expression upon drug sensitivity is currently unrecognized. PRP-4 belongs to the Ser/Thr protein kinase family, plays a role in pre-mRNA splicing and cell mitosis, and interacts with CLK1. Northern analysis demonstrates that PRP-4 is overexpressed in several paclitaxel resistant cell lines and confirms that PRP-4 expression could be significantly repressed by PRP-4 lentiviral shRNA. Both clonogenic and MTT assays confirm that transcriptional repression of PRP-4 could reverse paclitaxel resistance 5–10 fold in SKOV-3TR. Finally, overexpression of PRP-4 in drug sensitive cells could induce a modest level of drug resistance to paclitaxel, doxorubicin and vincristine.