title: Knockdown of CkrL by shRNA deteriorates hypoxia/reoxygenation-induced H9C2 cardiomyocyte apoptosis and survival inhibition Via Bax and downregulation of P-Erk1/2.
abstract: Integrin β1 subunit and its downstream molecule integrin-linked kinase and focal adhesion kinase have been confirmed to be essential to cell survival and inhibition of apoptosis and hypoxia/reoxygenation (H/R)-induced injuries in cardiomyocytes. However, it is still unclear whether CrkL [v-crk avian sarcoma virus CT-10 oncogene homolog (Crk)-like], which acts also as a component of the integrin pathway, could also affect H/R-induced injuries in the cardiomyocytes. The rat-derived H9C2 cardiomyocytes were infected with a CrkL small hairpin RNA interference recombinant lentivirus, which knockdowns the endogenous CrkL expression in the cardiomyocytes. Apoptosis, cell proliferation and survival were examined in the H9C2 cardiomyocytes treated with either H/R or not. Results showed that knockdown of CrkL could significantly increase apoptosis and inhibition of the cell proliferation and survival and deteriorate the previously mentioned injuries induced by H/R. In contrast, overexpression of human CrkL could relieve the exacerbation of the previously mentioned injuries induced by CrkL knockdown in the H9C2 cardiomyocytes via regulation of Bax and extracellular signal-regulated kinase1/2 (p-ERK1/2). In conclusion, these results confirmed that knockdown of CrkL could deteriorate H/R-induced apoptosis and cell survival inhibition in rat-derived H9C2 cardiomyocytes via Bax and downregulation of p-ERK1/2. It implies that CrkL could mitigate H/R-induced injuries in the cardiomyocytes.
authors.affiliation: Division of Cardiology, Department of Geriatrics, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China; Laboratory Research Center, The First Affiliated Hospital of Chongqing Medical University, Chongqing, China.
authors.forename: Zhi-Sheng
authors.initials: ZS
authors.lastname: Zhang
authors.forename: Dong-Yan
authors.initials: DY
authors.lastname: Yang
authors.forename: Yan-Bo
authors.initials: YB
authors.lastname: Fu
authors.forename: Lei
authors.initials: L
authors.lastname: Zhang
authors.forename: Qian-Ping
authors.initials: QP
authors.lastname: Zhao
authors.forename: Gang
authors.initials: G
authors.lastname: Li
mesh_terms: D000595:Amino Acid Sequence; D000818:Animals; D017209:Apoptosis; D015687:Cell Hypoxia; D002470:Cell Survival; D015536:Down-Regulation; D055785:Gene Knockdown Techniques; D006801:Humans; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism*; D008969:Molecular Sequence Data; D032383:Myocytes, Cardiac / Q000378:metabolism*; D050737:Proto-Oncogene Proteins c-crk / Q000235:genetics* / Q000378:metabolism*; D034741:RNA, Small Interfering / Q000378:metabolism*; D051381:Rats; D051028:bcl-2-Associated X Protein / Q000378:metabolism*
chemical_list: C485285:CRK protein, human; D050737:Proto-Oncogene Proteins c-crk; D034741:RNA, Small Interfering; D051028:bcl-2-Associated X Protein; D048052:Mitogen-Activated Protein Kinase 3
keywords: Bax; CrkL (v-crk avian sarcoma virus CT-10 oncogene homolog-like); H9C2 cardiomyocyte line; apoptosis; cell proliferation; cell survival; hypoxia/reoxygenation; phosphorylated extracellular signal-regulated kinase1/2 (p-ERK1/2)