title: Proteomic-based identification of Apg-2 as a therapeutic target for chronic myeloid leukemia.
abstract: The oncogenic BCR/ABL tyrosine kinase induces constitutive enhanced "spontaneous" DNA damage and unfaithful repair in Philadelphia chromosome positive leukemia cells. Here, we investigated the changes of protein profile in H2O2-induced DNA damage/repair in BaF3-MIGR1 and BaF3-BCR/ABL cells through a proteomic strategy consisting of two-dimensional gel electrophoresis (2-DE) coupled with MALDI-TOF mass spectrometry. In total, 41 spots were differentially expressed and 13 proteins were identified with further MS analysis. Two essential proteins, Proto-oncogene tyrosine-protein kinase ABL1 (c-ABL) and Heat shock 70kDa protein 4 (Apg-2), were confirmed by Western blot and showed consistent changes with proteomic results. Moreover, functional analysis demonstrated that inhibition of Apg-2 not only decreased cell proliferation, but also induced cell apoptosis in BCR/ABL positive cells (BaF3-BCR/ABL, BaF3-BCR/ABL(T315I)). We also proved that Apg-2 inhibition aggravated H2O2 induced damage in BCR/ABL positive cells, and enhanced the sensitivity of BaF3-BCR/ABL(T315I) to STI571. Taken together, the findings in this work provide us with some clues to a better understanding of the molecular mechanisms underlying BCR/ABL in the DNA damage/repair processes and demonstrated that Apg-2 would be a valid target for anti-leukemia drug development.
authors.affiliation: Department of Clinical Hematology, Key Laboratory of the Laboratory of Medical Diagnostics of the Ministry of Education, Chongqing Medical University, Chongqing 400016, PR China.
authors.forename: Yajuan
authors.initials: Y
authors.lastname: Li
authors.forename: Xi
authors.initials: X
authors.lastname: Chen
authors.forename: Meng
authors.initials: M
authors.lastname: Shi
authors.forename: Haixia
authors.initials: H
authors.lastname: Wang
authors.forename: Weixi
authors.initials: W
authors.lastname: Cao
authors.forename: Xiaozhong
authors.initials: X
authors.lastname: Wang
authors.forename: Chunli
authors.initials: C
authors.lastname: Li
authors.forename: Wenli
authors.initials: W
authors.lastname: Feng
mesh_terms: D000818:Animals; D000970:Antineoplastic Agents / Q000494:pharmacology; D001549:Benzamides / Q000494:pharmacology; D002453:Cell Cycle; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D019008:Drug Resistance, Neoplasm; D015972:Gene Expression Regulation, Neoplastic; D050957:HSP110 Heat-Shock Proteins / Q000235:genetics* / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000378:metabolism; D000068877:Imatinib Mesylate; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000188:drug therapy / Q000235:genetics* / Q000378:metabolism* / Q000473:pathology; D051379:Mice; D058990:Molecular Targeted Therapy; D010879:Piperazines / Q000494:pharmacology; D020543:Proteome / Q000378:metabolism; D040901:Proteomics; D011743:Pyrimidines / Q000494:pharmacology; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000235:genetics
chemical_list: D000970:Antineoplastic Agents; D001549:Benzamides; D050957:HSP110 Heat-Shock Proteins; C485680:Hspa4 protein, mouse; D010879:Piperazines; D020543:Proteome; D011743:Pyrimidines; D034741:RNA, Small Interfering; D000068877:Imatinib Mesylate; D006861:Hydrogen Peroxide
keywords: Apg-2; BCR/ABL; Chronic myeloid leukemia; H(2)O(2); Proteomic; shRNA