PMID- 27184254 OWN - NLM STAT- MEDLINE DCOM- 20170209 LR - 20191210 IS - 1757-2215 (Electronic) IS - 1757-2215 (Linking) VI - 9 IP - 1 DP - 2016 May 17 TI - HE4 promotes collateral resistance to cisplatin and paclitaxel in ovarian cancer cells. PG - 28 LID - 10.1186/s13048-016-0240-0 [doi] LID - 28 AB - BACKGROUND: Chemotherapy resistance presents a difficult challenge in treating epithelial ovarian cancer patients, particularly when tumors exhibit resistance to multiple chemotherapeutic agents. A few studies have shown that elevated serum levels of the ovarian cancer biomarker HE4 correlate with tumor chemoresistance, response to treatment, and survival. Here, we sought to confirm our previous results that HE4 contributes to collateral resistance to cisplatin and paclitaxel in vitro and uncover factors that may contribute to HE4-mediated chemoresistance. METHODS: MTS assays and western blots for cleaved PARP were used to assess resistance of HE4-overexpressing SKOV3 and OVCAR8 clones to cisplatin and paclitaxel. CRISPR/Cas technology was used to knockdown HE4 in HE4-overexpressing SKOV3 cells. A microarray was conducted to determine differential gene expression between SKOV3 null vector-transfected and HE4-overexpressing clones upon cisplatin exposure, and results were validated by quantitative RT-PCR. Regulation of mitogen activated protein kinases (MAPKs) and tubulins were assessed by western blot. RESULTS: HE4-overexpressing SKOV3 and OVCAR8 clones displayed increased resistance to cisplatin and paclitaxel. Knockdown of HE4 in HE4-overexpressing SKOV3 cells partially reversed chemoresistance. Microarray analysis revealed that HE4 overexpression resulted in suppression of cisplatin-mediated upregulation of EGR1, a MAPK-regulated gene involved in promoting apoptosis. Upregulation of p38, a MAPK activated in response to cisplatin, was suppressed in HE4-overexpressing clones. No differences in extracellular signal-regulated kinase (ERK) activation were noted in HE4-overexpressing clones treated with 25 muM cisplatin, but ERK activation was partially suppressed in HE4-overexpressing clones treated with 80 muM cisplatin. Furthermore, treatment of cells with recombinant HE4 dramatically affected ERK activation in SKOV3 and OVCAR8 wild type cells. Recombinant HE4 also upregulated alpha-tubulin and beta-tubulin levels in SKOV3 and OVCAR8 cells, and microtubule associated protein tau (MAPT) gene expression was increased in SKOV3 HE4-overexpressing clones. CONCLUSIONS: Overexpression of HE4 promotes collateral resistance to cisplatin and paclitaxel, and downregulation of HE4 partially reverses this chemoresistance. Multiple factors could be involved in HE4-mediated chemoresistance, including deregulation of MAPK signaling, as well as alterations in tubulin levels or stability. FAU - Ribeiro, J R AU - Ribeiro JR AD - Women and Infants Hospital, Department of Obstetrics and Gynecology, Program in Women's Oncology, Molecular Therapeutics Laboratory, 200 Chestnut Street, Providence, RI, 02903, USA. Jrribeiro@wihri.org. FAU - Schorl, C AU - Schorl C AD - Center for Genomics and Proteomics, Genomics Core Facility, Brown University, 70 Ship Street, Providence, RI, 02903, USA. FAU - Yano, N AU - Yano N AD - Women and Infants Hospital, Department of Obstetrics and Gynecology, Program in Women's Oncology, Molecular Therapeutics Laboratory, 200 Chestnut Street, Providence, RI, 02903, USA. FAU - Romano, N AU - Romano N AD - Women and Infants Hospital, Department of Obstetrics and Gynecology, Program in Women's Oncology, Molecular Therapeutics Laboratory, 200 Chestnut Street, Providence, RI, 02903, USA. FAU - Kim, K K AU - Kim KK AD - Wilmot Cancer Institute, Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, University of Rochester Medical Center, Rochester, NY, USA. FAU - Singh, R K AU - Singh RK AD - Women and Infants Hospital, Department of Obstetrics and Gynecology, Program in Women's Oncology, Molecular Therapeutics Laboratory, 200 Chestnut Street, Providence, RI, 02903, USA. AD - Wilmot Cancer Institute, Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, University of Rochester Medical Center, Rochester, NY, USA. FAU - Moore, R G AU - Moore RG AD - Women and Infants Hospital, Department of Obstetrics and Gynecology, Program in Women's Oncology, Molecular Therapeutics Laboratory, 200 Chestnut Street, Providence, RI, 02903, USA. AD - Wilmot Cancer Institute, Division of Gynecologic Oncology, Department of Obstetrics and Gynecology, University of Rochester Medical Center, Rochester, NY, USA. LA - eng GR - P30 GM114750/GM/NIGMS NIH HHS/United States PT - Journal Article DEP - 20160517 PL - England TA - J Ovarian Res JT - Journal of ovarian research JID - 101474849 RN - 0 (Antineoplastic Agents) RN - 0 (MAPT protein, human) RN - 0 (Proteins) RN - 0 (Tubulin) RN - 0 (WAP Four-Disulfide Core Domain Protein 2) RN - 0 (WFDC2 protein, human) RN - 0 (tau Proteins) RN - P88XT4IS4D (Paclitaxel) RN - Q20Q21Q62J (Cisplatin) SB - IM MH - Antineoplastic Agents/*pharmacology MH - Apoptosis/drug effects MH - CRISPR-Cas Systems MH - Cell Line, Tumor MH - Cell Proliferation/drug effects MH - Cell Survival/drug effects MH - Cisplatin/*pharmacology MH - Down-Regulation MH - Drug Resistance, Neoplasm/*genetics MH - Female MH - Gene Expression MH - Gene Expression Regulation, Neoplastic/drug effects MH - Humans MH - MAP Kinase Signaling System MH - Ovarian Neoplasms/*genetics/metabolism MH - Paclitaxel/*pharmacology MH - Proteins/*genetics MH - Tubulin/metabolism MH - WAP Four-Disulfide Core Domain Protein 2 MH - tau Proteins/genetics/metabolism PMC - PMC4869286 OTO - NOTNLM OT - Chemoresistance OT - EGR1 OT - Epithelial ovarian cancer OT - HE4 OT - MAPK signaling OT - Paclitaxel resistance OT - Platinum resistance OT - SKOV3 cells OT - Tubulin EDAT- 2016/05/18 06:00 MHDA- 2017/02/10 06:00 PMCR- 2016/05/17 CRDT- 2016/05/18 06:00 PHST- 2016/03/07 00:00 [received] PHST- 2016/05/05 00:00 [accepted] PHST- 2016/05/18 06:00 [entrez] PHST- 2016/05/18 06:00 [pubmed] PHST- 2017/02/10 06:00 [medline] PHST- 2016/05/17 00:00 [pmc-release] AID - 10.1186/s13048-016-0240-0 [pii] AID - 240 [pii] AID - 10.1186/s13048-016-0240-0 [doi] PST - epublish SO - J Ovarian Res. 2016 May 17;9(1):28. doi: 10.1186/s13048-016-0240-0.