Overexpression of BCL-XL Underlies the Molecular Basis for Resistance to Staurosporine-induced Apoptosis in PC-3 Cells

We have reported previously that among human prostate cancer cell lines LNCaP but not PC-3 cells undergo apoptosis after treatment with the protein kinase inhibitor staurosporine (STS). We have now further investigated this model to uncover the molecular mechanism causing resistance to STS-induced apoptosis in PC-3 cells. S-100 lysates of both cell lines showed biochemical changes typical of apoptosis after the addition of cytochrome c and dATP, suggesting that the postmitochondrial phase of apoptosis was intact. Upon addition of STS, the proapoptotic molecules Bax and Bad became predominantly mitochondrial in both cell lines. This, in turn, was followed by loss of mitochondrial transmembrane potential, translocation of cytochrome c to the cytosol, activation of caspase-9, -3, and -7, and cleavage of the apoptotic targets, DNA fragmentation factor and poly(ADP-ribose) polymerase, in LNCaP but not in PC-3 cells. Components of the mitochondrial permeability transition pore, adenine nucleotide transporter and voltage-dependent anion channel, were normally expressed in the correct subcellular fraction of both cell lines. Overexpression of the proapoptotic proteins Bax and Bad, fused to a green fluorescent protein but not of green fluorescent protein alone, induced apoptosis in >80% of PC-3 cells. These experiments suggested that a factor protecting the mitochondria of PC-3 cells mediates resistance to STS-induced apoptosis. A wide search among the antiapoptotic Bcl-2 family members was performed, and Bcl-XL was found to be overexpresseAd in PC-3 cells. Experiments down-regulating Bcl-XL expression by using the tyrosine kinase inhibitor genistein, sodium butyrate, or an antisense Bcl-XL oligonucleotide restored sensitivity to apoptosis in PC-3 cells. Thus, Bcl-XL overexpression is one of the mediators of resistance to STS-induced apoptosis in the prostate cancer cell line PC-3.

[1]  Robert L Moritz,et al.  Identification of DIABLO, a Mammalian Protein that Promotes Apoptosis by Binding to and Antagonizing IAP Proteins , 2000, Cell.

[2]  Xiaodong Wang,et al.  Smac, a Mitochondrial Protein that Promotes Cytochrome c–Dependent Caspase Activation by Eliminating IAP Inhibition , 2000, Cell.

[3]  R. Bernards,et al.  Distinct Initiation and Maintenance Mechanisms Cooperate to Induce G1 Cell Cycle Arrest in Response to DNA Damage , 2000, Cell.

[4]  Xiaodong Wang,et al.  Cytochrome c Deficiency Causes Embryonic Lethality and Attenuates Stress-Induced Apoptosis , 2000, Cell.

[5]  Y Li,et al.  [Mitochondria and apoptosis]. , 2000, Zhonghua yu fang yi xue za zhi [Chinese journal of preventive medicine].

[6]  M. Marcelli,et al.  Heterogeneous apoptotic responses of prostate cancer cell lines identify an association between sensitivity to staurosporine‐induced apoptosis, expression of Bcl‐2 family members, and caspase activation , 2000, The Prostate.

[7]  J. Martinou,et al.  Bid Induces the Oligomerization and Insertion of Bax into the Outer Mitochondrial Membrane , 2000, Molecular and Cellular Biology.

[8]  Zhi-Min Yuan,et al.  Translocation of SAPK/JNK to Mitochondria and Interaction with Bcl-xL in Response to DNA Damage* , 2000, The Journal of Biological Chemistry.

[9]  T. Denning,et al.  Acquired alkylating drug resistance of a human ovarian carcinoma cell line is unaffected by altered levels of pro- and anti-apoptotic proteins , 2000, Oncogene.

[10]  Junying Yuan,et al.  Caspase-12 mediates endoplasmic-reticulum-specific apoptosis and cytotoxicity by amyloid-β , 2000, Nature.

[11]  P. Walsh,et al.  Early androgen deprivation for prostate cancer? , 1999, The New England journal of medicine.

[12]  G. Zhou,et al.  c-Jun N-terminal kinase mediates apoptotic signaling induced by N-(4-hydroxyphenyl)retinamide. , 1999, Molecular pharmacology.

[13]  A. Bielawska,et al.  Aberrant sphingolipid signaling is involved in the resistance of prostate cancer cell lines to chemotherapy. , 1999, Cancer research.

[14]  K. Arai,et al.  Two distinct signaling pathways downstream of Janus kinase 2 play redundant roles for antiapoptotic activity of granulocyte-macrophage colony-stimulating factor. , 1999, Molecular biology of the cell.

[15]  R. Jove,et al.  Inhibition of pp60c-Src reduces Bcl-XL expression and reverses the transformed phenotype of cells overexpressing EGF and HER-2 receptors , 1999, Oncogene.

[16]  Y. Hsu,et al.  Conformation of the Bax C‐terminus regulates subcellular location and cell death , 1999, The EMBO journal.

[17]  C. F. Bennett,et al.  The Role of Antiapoptotic Bcl-2 Family Members in Endothelial Apoptosis Elucidated with Antisense Oligonucleotides* , 1999, The Journal of Biological Chemistry.

[18]  M. Greenberg,et al.  Akt Promotes Cell Survival by Phosphorylating and Inhibiting a Forkhead Transcription Factor , 1999, Cell.

[19]  B. Corfe,et al.  Cell Damage-induced Conformational Changes of the Pro-Apoptotic Protein Bak In Vivo Precede the Onset of Apoptosis , 1999, The Journal of cell biology.

[20]  Jean-Claude Martinou,et al.  Bid-induced Conformational Change of Bax Is Responsible for Mitochondrial Cytochrome c Release during Apoptosis , 1999, The Journal of cell biology.

[21]  W. Zong,et al.  The prosurvival Bcl-2 homolog Bfl-1/A1 is a direct transcriptional target of NF-κB that blocks TNFα-induced apoptosis , 1999 .

[22]  Ruedi Aebersold,et al.  Molecular characterization of mitochondrial apoptosis-inducing factor , 1999, Nature.

[23]  M. V. Heiden,et al.  Bcl-xL prevents cell death following growth factor withdrawal by facilitating mitochondrial ATP/ADP exchange. , 1999, Molecular cell.

[24]  M. Marcelli,et al.  Signaling pathway activated during apoptosis of the prostate cancer cell line LNCaP: overexpression of caspase-7 as a new gene therapy strategy for prostate cancer. , 1999, Cancer research.

[25]  S. Korsmeyer,et al.  Caspase Cleaved BID Targets Mitochondria and Is Required for Cytochrome c Release, while BCL-XL Prevents This Release but Not Tumor Necrosis Factor-R1/Fas Death* , 1999, The Journal of Biological Chemistry.

[26]  T. Chittenden,et al.  Bax interacts with the permeability transition pore to induce permeability transition and cytochrome c release in isolated mitochondria. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[27]  John Calvin Reed,et al.  Regulation of cell death protease caspase-9 by phosphorylation. , 1998, Science.

[28]  S. Korsmeyer,et al.  Regulated Targeting of BAX to Mitochondria , 1998, The Journal of cell biology.

[29]  J C Reed,et al.  Bax and adenine nucleotide translocator cooperate in the mitochondrial control of apoptosis. , 1998, Science.

[30]  C. Y. Wang,et al.  NF-kappaB antiapoptosis: induction of TRAF1 and TRAF2 and c-IAP1 and c-IAP2 to suppress caspase-8 activation. , 1998, Science.

[31]  V. Dixit,et al.  Death receptors: signaling and modulation. , 1998, Science.

[32]  Y. Lazebnik,et al.  Caspases: enemies within. , 1998, Science.

[33]  Xiaodong Wang,et al.  Bid, a Bcl2 Interacting Protein, Mediates Cytochrome c Release from Mitochondria in Response to Activation of Cell Surface Death Receptors , 1998, Cell.

[34]  Junying Yuan,et al.  Cleavage of BID by Caspase 8 Mediates the Mitochondrial Damage in the Fas Pathway of Apoptosis , 1998, Cell.

[35]  J. Cleveland,et al.  Selective regulation of Bcl-XL by a Jak kinase-dependent pathway is bypassed in murine hematopoietic malignancies. , 1998, Genes & development.

[36]  S. Korsmeyer,et al.  Enforced dimerization of BAX results in its translocation, mitochondrial dysfunction and apoptosis , 1998, The EMBO journal.

[37]  E. Alnemri,et al.  Abrogation of Mitochondrial Cytochrome c Release and Caspase-3 Activation in Acquired Multidrug Resistance* , 1998, The Journal of Biological Chemistry.

[38]  C. Rudin,et al.  A Bcl-xL transgene promotes malignant conversion of chemically initiated skin papillomas. , 1998, Cancer research.

[39]  W. Cavenee,et al.  Drug resistance of human glioblastoma cells conferred by a tumor-specific mutant epidermal growth factor receptor through modulation of Bcl-XL and caspase-3-like proteases. , 1998, Proceedings of the National Academy of Sciences of the United States of America.

[40]  S. Srinivasula,et al.  Cytochrome c and dATP-Dependent Formation of Apaf-1/Caspase-9 Complex Initiates an Apoptotic Protease Cascade , 1997, Cell.

[41]  C. Stein,et al.  Taxol and estramustine-induced modulation of human prostate cancer cell apoptosis via alteration in bcl-xL and bak expression. , 1997, Clinical cancer research : an official journal of the American Association for Cancer Research.

[42]  L. Peso,et al.  Interleukin-3-induced phosphorylation of BAD through the protein kinase Akt. , 1997, Science.

[43]  John Calvin Reed,et al.  IFN-gamma upregulates anti-apoptotic gene expression and inhibits apoptosis in IL-3-dependent hematopoietic cells. , 1997, Biochemical and biophysical research communications.

[44]  S. R. Datta,et al.  Akt Phosphorylation of BAD Couples Survival Signals to the Cell-Intrinsic Death Machinery , 1997, Cell.

[45]  R. Weichselbaum,et al.  Role for Bcl-xL as an inhibitor of cytosolic cytochrome C accumulation in DNA damage-induced apoptosis. , 1997, Proceedings of the National Academy of Sciences of the United States of America.

[46]  Elizabeth Yang,et al.  Serine Phosphorylation of Death Agonist BAD in Response to Survival Factor Results in Binding to 14-3-3 Not BCL-XL , 1996, Cell.

[47]  Xiaodong Wang,et al.  Induction of Apoptotic Program in Cell-Free Extracts: Requirement for dATP and Cytochrome c , 1996, Cell.

[48]  M. Mandal,et al.  Overexpression of HER2 modulates bcl-2, bcl-XL, and tamoxifen-induced apoptosis in human MCF-7 breast cancer cells. , 1996, Clinical cancer research : an official journal of the American Association for Cancer Research.

[49]  D. Louis,et al.  The BAX gene maps to the glioma candidate region at 19q13.3, but is not altered in human gliomas. , 1996, Cancer genetics and cytogenetics.

[50]  Z. Oltvai,et al.  Multiple Bcl-2 family members demonstrate selective dimerizations with Bax. , 1995, Proceedings of the National Academy of Sciences of the United States of America.

[51]  G. Kroemer,et al.  Sequential reduction of mitochondrial transmembrane potential and generation of reactive oxygen species in early programmed cell death , 1995, The Journal of experimental medicine.

[52]  Z. Oltvai,et al.  BH1 and BH2 domains of Bcl-2 are required for inhibition of apoptosis and heterodimerization with Bax , 1994, Nature.

[53]  C. Thompson,et al.  bcl-x, a bcl-2-related gene that functions as a dominant regulator of apoptotic cell death , 1993, Cell.

[54]  D. Chopin,et al.  Detection of the apoptosis-suppressing oncoprotein bc1-2 in hormone-refractory human prostate cancers. , 1993, The American journal of pathology.

[55]  M. Campbell,et al.  Expression of the protooncogene bcl-2 in the prostate and its association with emergence of androgen-independent prostate cancer. , 1992, Cancer research.

[56]  N. Kyprianou,et al.  Relationship between metastatic ability and H-ras oncogene expression in rat mammary cancer cells transfected with the v-H-ras oncogene. , 1990, Cancer research.

[57]  J. Lechner,et al.  Establishment and characterization of a human prostatic carcinoma cell line (PC-3). , 1979, Investigative urology.

[58]  M. Marcelli,et al.  Caspase-7 is activated during lovastatin-induced apoptosis of the prostate cancer cell line LNCaP. , 1998, Cancer research.

[59]  S. Arya,et al.  The LNCaP cell line--a new model for studies on human prostatic carcinoma. , 1980, Progress in clinical and biological research.