title: [Establishment of cisplatin-resistant human endometrial cancer cell line and the study of its resistant mechanisms].
abstract: OBJECTIVE
  To establish the cisplatin (DDP)-resistant cell line from human endometrial cancer cell line Ishikawa and to investigate its resistant mechanism to DDP.
  
  
  METHODS
  A resistant endometrial cancer cell line ISH/DDP was established by gradually increasing dose of cisplatin and high-dose stimulation. The resistant index was estimated by 3-(4,5-dimethylthiazol-zyl)-5-(3-carboxymethoxyphenyl-2-(4-sulfophenyl)-2H-tetrazolium, inner salt (MTS) assay. Cell growth curve, doubling time and cell cycle phase distribution were measured; drug-resistant protein of breast cancer resistance protein (BCRP), P-glycoprotein (P-gp) and glutathione-S-transferase-π (GST-π) were examined by immuocytochemistry. Results The DDP-resistant cell line ISH/DDP was established with the resistant index of 3.77. The proliferation of ISH/DDP got slow, doubling time prolonged, which were 40.1 hours, while it was 34.1 hours in Ishikawa (P<0.05); and the cell number of G0/G1 phase [(44.3±5.7)% and (39.0±10.7)%, P>0.05] and G2/M phase [(11.9±0.7)% and (5.7±2.4)%, P<0.05] decreased, while S-phase [(44.2±6.1)% and (55.3±8.4)%, P<0.05] increased compared with parent cells. The comprehensive score of the expression of BCRP in ISH/DDP was 16.3±2.0, while it was 13.4±1.5 in Ishikawa (P<0.05). The score of the expression of P-gp in ISH/DDP and Ishikawa were 15.5±1.2 and 16.1±1.0 (P>0.05), respectively. The score of the expression of GST-π in ISH/DDP and Ishikawa were 15.2±1.9 and 14.9±1.1 (P>0.05).
  
  
  CONCLUSIONS
  ISH/DDP cell line showed a typical resistant phenotype and biological characteristics, which may be accounted for high BCRP expression.
authors: Department of Obstetrics and Gynecology, Peking University People's Hospital, Beijing 100044, China.
authors: Qi-ying
authors: QY
authors: Xu
authors: Yong-hua
authors: YH
authors: Chen
authors: Li-jun
authors: LJ
authors: Zhao
authors: Xiao-ping
authors: XP
authors: Li
authors: Jian-liu
authors: JL
authors: Wang
authors: Li-hui
authors: LH
authors: Wei
mesh_terms: D018435:ATP Binding Cassette Transporter, Subfamily B / Q000378:metabolism; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters / Q000378:metabolism*; D000970:Antineoplastic Agents / Q000008:administration & dosage / Q000494:pharmacology*; D002453:Cell Cycle / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002945:Cisplatin / Q000008:administration & dosage / Q000494:pharmacology*; D019008:Drug Resistance, Neoplasm; D016889:Endometrial Neoplasms / Q000378:metabolism / Q000473:pathology*; D005260:Female; D005434:Flow Cytometry; D051549:Glutathione S-Transferase pi / Q000378:metabolism; D006801:Humans; D009363:Neoplasm Proteins / Q000378:metabolism*
chemical_list: C116614:ABCG2 protein, human; D018435:ATP Binding Cassette Transporter, Subfamily B; D000070997:ATP Binding Cassette Transporter, Subfamily G, Member 2; D018528:ATP-Binding Cassette Transporters; D000970:Antineoplastic Agents; D009363:Neoplasm Proteins; D051549:Glutathione S-Transferase pi; D002945:Cisplatin