RRC ID 59904
著者 Wang-Bishop L, Chen Z, Gomaa A, Lockhart AC, Salaria S, Wang J, Lewis KB, Ecsedy J, Washington K, Beauchamp RD, El-Rifai W.
タイトル Inhibition of AURKA Reduces Proliferation and Survival of Gastrointestinal Cancer Cells With Activated KRAS by Preventing Activation of RPS6KB1.
ジャーナル Gastroenterology
Abstract BACKGROUND & AIMS:Activation of KRAS signaling and overexpression of the aurora kinase A (AURKA) are often detected in luminal gastrointestinal cancers. We investigated regulation of ribosomal protein S6 kinase B1 (RPS6KB1) by AURKA and the effects of alisertib, an AURKA inhibitor, in mice xenograft tumors grown from human gastrointestinal cancer cells with mutant, activated forms of KRAS.
METHODS:We tested the effects of alisertib or AURKA overexpression or knockdown in 10 upper gastrointestinal or colon cancer cell lines with KRAS mutations or amplifications using the CellTiter-Glo luminescence and clonogenic cell survival assays. We used the proximity ligation in situ assay to evaluate protein co-localization and immunoprecipitation to study protein interactions. Nude mice with xenograft tumors grown from HCT116, SNU-601, SW480, or SNU-1 cells were given oral alisertib (40 mg/kg, 5 times/wk) for 4 weeks. Tumor samples were collected and analyzed by immunoblots and immunohistochemistry. Tissue microarrays from 151 paraffin-embedded human colon tumors, with adjacent normal and adenoma tissues, were analyzed by immunohistochemistry for levels of AURKA.
RESULTS:Alisertib reduced proliferation and survival of the cell lines tested. AURKA knockdown or inhibition with alisertib reduced levels of phosphorylated RPS6KB1 (at T389) and increased levels of proteins that induce apoptosis, including BIM, cleaved PARP, and cleaved caspase 3. AURKA co-localized and interacted with RPS6KB1, mediating RPS6KB1 phosphorylation at T389. We detected AURKA-dependent phosphorylation of RPS6KB1 in cell lines with mutations in KRAS but not in cells with wild-type KRAS. Administration of alisertib to mice with xenograft tumors significantly reduced tumor volumes (P < .001). Alisertib reduced phosphorylation of RPS6KB1 and Ki-67 and increased levels of cleaved caspase 3 in tumor tissues. In analyses of tissue microarrays, we found significant overexpression of AURKA in gastrointestinal tumor tissues compared with non-tumor tissues (P = .0003).
CONCLUSION:In studies of gastrointestinal cancer cell lines with activated KRAS, we found AURKA to phosphorylate RPS6KB1, promoting cell proliferation and survival and growth of xenograft tumors in mice. Agents that inhibit AURKA might slow the growth of gastrointestinal tumors with activation of KRAS.
巻・号 156(3)
ページ 662-675.e7
公開日 2019-2-1
DOI 10.1053/j.gastro.2018.10.030
PII S0016-5085(18)35161-8
PMID 30342037
PMC PMC6368861
MeSH Animals Aurora Kinase A / antagonists & inhibitors* Aurora Kinase A / genetics Aurora Kinase A / pharmacology Azepines / pharmacology* Cell Death / drug effects Cell Death / genetics Cell Line, Tumor Cell Proliferation / drug effects Cell Proliferation / genetics Gastrointestinal Neoplasms / drug therapy Gastrointestinal Neoplasms / genetics* Gastrointestinal Neoplasms / pathology Gene Expression Regulation, Neoplastic Humans Mice Mice, Nude Molecular Targeted Therapy Protein Kinase Inhibitors / pharmacology Proto-Oncogene Proteins p21(ras) / genetics* Pyrimidines / pharmacology* Random Allocation Sensitivity and Specificity Signal Transduction / genetics Xenograft Model Antitumor Assays
IF 19.809
引用数 11
リソース情報
ヒト・動物細胞 HCT116(RCB2979) MKN45(RCB1001)