title: p21CIP1 Induces Apoptosis via Binding to BCL2 in LNCaP Prostate Cancer Cells Treated with MCS-C3, A Novel Carbocyclic Analog of Pyrrolopyrimidine.
abstract: BACKGROUND
  Previously, we synthesized a new carbocyclic analog of pyrrolo[2,3-d]pyrimidine nucleoside, designated MCS-C3. Recently, we found that LNCaP androgen-responsive prostate cancer cells treated with MCS-C3 rapidly undergo intrinsic apoptosis through dramatic up-regulation of p21(CIP1). The present study aimed to evaluate the cellular functions and underlying molecular mechanisms of p21(CIP1) on apoptotic induction in LNCaP cells treated with 6 μM MCS-C3.
  
  
  METHODS
  Western blots, flow cytometric assay, immunoprecipitation, and transmission electron microscopy analysis were used to measure apoptotic induction in 6-μM MCS-C3-treated LNCaP cells. Effects of MCS-C3 on gene expression of p21(CIP1) were measured by semi-quantitative real-time polymerase chain reaction, and small interfering RNA transfection.
  
  
  RESULTS
  MCS-C3 induced appreciable caspase-dependent apoptosis associated with the significant up-regulation of p53-dependent p21(CIP1) in LNCaP cells. Moreover, this apoptotic induction was caused by direct binding of p21(CIP1) to anti-apoptotic B-cell lymphoma 2 (BCL2) protein, and antagonizing BCL2 function. In addition, MCS-C3-mediated apoptotic induction, and up-regulation of p21(CIP1) were almost completely blocked by the treatment of androgen-responsive LNCaP cells with flutamide, an androgen receptor (AR) antagonist.
  
  
  CONCLUSIONS
  We identified that induction of intrinsic apoptosis in LNCaP cells by 6 μM MCS-C3 is associated not only with p53 activation but also with mediation of AR. In the present study, we identified the cellular functions and underlying molecular mechanisms of p53-dependent and AR-associated p21(CIP1) on apoptotic induction via direct binding to BCL2 in LNCaP cells treated with 6 μM MCS-C3.
authors.affiliation: Department of Pharmacy, College of Pharmacy and Institute of Pharmaceutical Science and Technology, Hanyang University, Gyeonggi-do, Republic of Korea.
authors.forename: Ko-Woon
authors.initials: KW
authors.lastname: Choi
authors.affiliation: Department of Pharmacy, College of Pharmacy and Institute of Pharmaceutical Science and Technology, Hanyang University, Gyeonggi-do, Republic of Korea.
authors.forename: Hyewon
authors.initials: H
authors.lastname: Suh
authors.affiliation: Department of Pharmacy, College of Pharmacy and Institute of Pharmaceutical Science and Technology, Hanyang University, Gyeonggi-do, Republic of Korea.
authors.forename: Ha Lim
authors.initials: HL
authors.lastname: Oh
authors.affiliation: Department of Pharmacy, College of Pharmacy and Institute of Pharmaceutical Science and Technology, Hanyang University, Gyeonggi-do, Republic of Korea.
authors.forename: Chongsuk
authors.initials: C
authors.lastname: Ryou
authors.affiliation: Department of Pharmacy, College of Pharmacy and Institute of Pharmaceutical Science and Technology, Hanyang University, Gyeonggi-do, Republic of Korea chhlee@hanyang.ac.kr.
authors.forename: Chul-Hoon
authors.initials: CH
authors.lastname: Lee
mesh_terms: D017209:Apoptosis; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D006801:Humans; D008297:Male; D011471:Prostatic Neoplasms / Q000188:drug therapy* / Q000473:pathology; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000276:immunology*; D011743:Pyrimidines / Q000378:metabolism*; D011758:Pyrroles / Q000378:metabolism*; D015398:Signal Transduction; D014162:Transfection
chemical_list: C000595707:BCL2 protein, human; D019253:Proto-Oncogene Proteins c-bcl-2; D011743:Pyrimidines; D011758:Pyrroles; C527741:pyrrolopyrimidine
keywords: BCL2; LNCaP cells; Prostate cancer; apoptosis; p21CIP1