Inhibition of Girdin enhances chemosensitivity of colorectal cancer cells to oxaliplatin

• Published in: World journal of gastroenterology : WJG 2014-07, Vol.20 (25), p.8229-8236
• Main Authors: Zhang, Ya-Jie, Li, A-Jian, Han, Yi, Yin, Lu, Lin, Mou-Bin
• Format: Article
• Language: English
• Subjects: Antineoplastic Combined Chemotherapy Protocols - pharmacology; Caco-2 Cells; can; Cell Death - drug effects; Cell Proliferation - drug effects; Chemosensitivity; Clinical Trials Study; Colorectal; Colorectal Neoplasms - genetics; Colorectal Neoplasms - metabolism; Colorectal Neoplasms - pathology; DNA Topoisomerases, Type II - genetics; DNA Topoisomerases, Type II - metabolism; DNA-Binding Proteins - antagonists & inhibitors; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; Dose-Response Relationship, Drug; Down-Regulation; Doxorubicin - pharmacology; Drug Resistance, Neoplasm - genetics; Gene Expression Regulation, Neoplastic; Girdin; HCT116 Cells; Humans; Microfilament Proteins - genetics; Microfilament Proteins - metabolism; Organoplatinum Compounds - pharmacology; Oxaliplatin; Poly-ADP-Ribose Binding Proteins; RNA Interference; RNA, Messenger - metabolism; Topoisomerase II Inhibitors - pharmacology; Transfection; Vesicular Transport Proteins - genetics; Vesicular Transport Proteins - metabolism
• ISSN: 1007-9327; 2219-2840
• DOI: 10.3748/wjg.v20.i25.8229

AIM: 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 Girdinexpression 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).CONCLUSION: 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.