title: Inhibition of Girdin enhances chemosensitivity of colorectal cancer cells to oxaliplatin.
abstract: OBJECTIVE
  To investigate the effect of Girdin knockdown on the chemosensitivity of colorectal cancer cells to oxaliplatin and the possible mechanisms involved.
  
  
  METHODS
  Four siRNAs targeting Girdin were transfected into the chemoresistant colorectal cancer cell line DLD1. Real-time polymerase chain reaction (PCR) was employed to assess Girdin mRNA expression and the most effective siRNA was chosen for conversion into shRNA. Then, DLD1 cells were infected with lentiviruses expressing the Girdin shRNA and a scramble control, respectively, and Girdin mRNA and protein expression levels were assessed by real-time PCR and Western blotting. Furthermore, microarray experiments were used to assess global gene expression profile after Girdin suppression in DLD1 cells. Finally, the cytotoxic effect of simultaneous treatment with oxaliplatin and adriamycin (an inhibitor of a significantly downregulated gene after Girdin suppression in DLD1 cells) was examined by MTT assay.
  
  
  RESULTS
  The most effective siRNA suppressed Girdin expression with an inhibition efficiency of 57%. Compared with the scramble control, DLD1 cells infected with the Girdin shRNA displayed decreased Girdin mRNA and protein levels (P < 0.05), and Girdin knockdown significantly enhanced chemosensitivity to oxaliplatin in colorectal cancer cells (P < 0.05). Microarray data revealed that 381 and 162 genes were upregulated and downregulated in response to Girdin reduction, respectively, with ratios > 1.2 or < 0.8 (P < 0.01). Interestingly, TOP2B (DNA topoisomerase 2-β) was downregulated (ratio = 0.78, P = 0.0001) and oxaliplatin/adriamycin combination resulted in increased cell death compared with treatments with individual agents (P < 0.05).
  
  
  CONCLUSIONS
  Girdin knockdown enhances chemosensitivity of colorectal cancer cells to oxaliplatin via TOP2B down-regulation. These findings provide a promising approach to overcome the chemoresistance of colorectal cancer cells.
authors.affiliation: Ya-Jie Zhang, A-Jian Li, Yi Han, Lu Yin, Mou-Bin Lin, Department of General Surgery, RuiJin Hospital Affiliated Shanghai Jiaotong University School of Medicine, Shanghai 200025, China.
authors.forename: Ya-Jie
authors.initials: YJ
authors.lastname: Zhang
authors.affiliation: Ya-Jie Zhang, A-Jian Li, Yi Han, Lu Yin, Mou-Bin Lin, Department of General Surgery, RuiJin Hospital Affiliated Shanghai Jiaotong University School of Medicine, Shanghai 200025, China.
authors.forename: A-Jian
authors.initials: AJ
authors.lastname: Li
authors.affiliation: Ya-Jie Zhang, A-Jian Li, Yi Han, Lu Yin, Mou-Bin Lin, Department of General Surgery, RuiJin Hospital Affiliated Shanghai Jiaotong University School of Medicine, Shanghai 200025, China.
authors.forename: Yi
authors.initials: Y
authors.lastname: Han
authors.affiliation: Ya-Jie Zhang, A-Jian Li, Yi Han, Lu Yin, Mou-Bin Lin, Department of General Surgery, RuiJin Hospital Affiliated Shanghai Jiaotong University School of Medicine, Shanghai 200025, China.
authors.forename: Lu
authors.initials: L
authors.lastname: Yin
authors.affiliation: Ya-Jie Zhang, A-Jian Li, Yi Han, Lu Yin, Mou-Bin Lin, Department of General Surgery, RuiJin Hospital Affiliated Shanghai Jiaotong University School of Medicine, Shanghai 200025, China.
authors.forename: Mou-Bin
authors.initials: MB
authors.lastname: Lin
mesh_terms: D000971:Antineoplastic Combined Chemotherapy Protocols / Q000494:pharmacology*; D018938:Caco-2 Cells; D016923:Cell Death / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D015179:Colorectal Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology; D004250:DNA Topoisomerases, Type II / Q000235:genetics / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D015536:Down-Regulation; D004317:Doxorubicin / Q000494:pharmacology; D019008:Drug Resistance, Neoplasm / Q000235:genetics; D015972:Gene Expression Regulation, Neoplastic; D045325:HCT116 Cells; D006801:Humans; D008840:Microfilament Proteins / Q000235:genetics / Q000378:metabolism*; D009944:Organoplatinum Compounds / Q000494:pharmacology*; D000077150:Oxaliplatin; D000075223:Poly-ADP-Ribose Binding Proteins; D034622:RNA Interference; D012333:RNA, Messenger / Q000378:metabolism; D059005:Topoisomerase II Inhibitors / Q000494:pharmacology*; D014162:Transfection; D033921:Vesicular Transport Proteins / Q000235:genetics / Q000378:metabolism*
chemical_list: C502451:CCDC88A protein, human; D004268:DNA-Binding Proteins; D008840:Microfilament Proteins; D009944:Organoplatinum Compounds; D000075223:Poly-ADP-Ribose Binding Proteins; D012333:RNA, Messenger; D059005:Topoisomerase II Inhibitors; D033921:Vesicular Transport Proteins; D000077150:Oxaliplatin; D004317:Doxorubicin; D004250:DNA Topoisomerases, Type II; C000618814:TOP2B protein, human
keywords: Chemosensitivity; Colorectal cancer; Girdin; Oxaliplatin; TOP2B