title: Cisplatin treatment leads to changes in nuclear protein and microRNA expression.
abstract: Using a proteomic approach, we have previously shown that exposure to different concentrations of cisplatin during a 12-h period can lead to changes in nuclear protein expression and alternative splicing in HeLa cells. To further shed light on the DNA damage response (DDR) induced by cisplatin, we examined the nuclear proteome profiles of HeLa cells treated with 5μM cisplatin for different times (2, 12, and 24h). Two-dimensional electrophoresis (2-DE) identified 98 differentially expressed proteins in cisplatin-treated cells as compared to control cells. Among them, 54 spots (55%) were down-regulated and 44 spots (45%) were up-regulated. 51 spots were subjected to Matrix-assisted-laser-desorption-ionization Time-of-flight/time-of-flight Mass spectrometry (MALDI-TOF/TOF MS) identification, and 40 spots were identified. Among these, 22 proteins were located in nucleus. These proteins were involved in stress response, cell cycle and division, apoptosis, mRNA processing, transport, splicing and microRNA (miRNA) maturation. The changed expression of Annexin A1 and Lamin B1 were confirmed by Western blot. The role of Annexin A1 in the response to cisplatin-induced DNA damage was further analyzed, and it was shown that after Annexin A1 knockdown, cisplatin-induced DNA damage was significantly increased. In addition, the changed expression of several miRNAs was also observed by quantitative real-time PCR (qRT-PCR). Taken together, these data indicate that cisplatin-induced DDR is a complex process, and that those proteins identified by proteomics can lead to new directions for a better understanding of this process.
authors.affiliation: The First Affiliated Hospital, Zhejiang University, Hangzhou, Zhejiang, China.
authors.forename: Guanglin
authors.initials: G
authors.lastname: Zhang
authors.forename: Li
authors.initials: L
authors.lastname: Sun
authors.forename: Xianghong
authors.initials: X
authors.lastname: Lu
authors.forename: Zhanghui
authors.initials: Z
authors.lastname: Chen
authors.forename: Penelope J
authors.initials: PJ
authors.lastname: Duerksen-Hughes
authors.forename: Hu
authors.initials: H
authors.lastname: Hu
authors.forename: Xinqiang
authors.initials: X
authors.lastname: Zhu
authors.forename: Jun
authors.initials: J
authors.lastname: Yang
mesh_terms: D017305:Annexin A1 / Q000235:genetics; D000970:Antineoplastic Agents / Q000633:toxicity*; D002470:Cell Survival / Q000187:drug effects; D002945:Cisplatin / Q000633:toxicity*; D004249:DNA Damage / Q000187:drug effects*; D004305:Dose-Response Relationship, Drug; D055785:Gene Knockdown Techniques; D006367:HeLa Cells; D006801:Humans; D035683:MicroRNAs / Q000187:drug effects* / Q000378:metabolism; D009687:Nuclear Proteins / Q000187:drug effects / Q000378:metabolism*; D014162:Transfection
chemical_list: D017305:Annexin A1; D000970:Antineoplastic Agents; D035683:MicroRNAs; D009687:Nuclear Proteins; D002945:Cisplatin