Qing Chang1, Renyi Qin, Tao Huang, Jun Gao, Yanping Feng. 1. Department of Surgery, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei Province, China.
Abstract
OBJECTIVES: The aim of the study was to observe the effect of antisense hypoxia-inducible factor 1alpha (HIF-1alpha) on progression, metastasis, and chemosensitivity of pancreatic cancer. METHODS: BxPc-3 cells transfected with antisense HIF-1alpha plasmid were exposed to 0.5% O2 for 4 hours. Expressions of HIF-1alpha, survivin, and beta1 integrin were detected by reverse transcriptase -polymerase chain reaction and Western blotting. Growth inhibition rates and apoptosis rates of BxPc-3 cells under different dosages of chemotherapy agents (5-fluorouracil, doxorubicin, and gemcitabine) were measured by MTT colorimetric assay and flow cytometry. The migration of BxPc-3 cells was assayed using transwell cell culture chambers. Subcutaneous transplantation of BxPc-3 cells in nude mice for 8 weeks was to assess progression and metastasis of pancreatic cancer. RESULTS: Expression of HIF-1alpha was obviously down-regulated, and at the same time, survivin and beta1-integrin expressions were markedly down-regulated in the experimental group (P < 0.05). Higher dosages (100, 200, and 400 mg/L of 5-fluorouracil; 0.05, 0.075, and 0.1 mg/L of doxorubicin; and 10(-9), 10(-8), and 10(-7) mol/L of gemcitabine) caused a greater increase of inhibition in the experimental group than in control (P < 0.05). The number of migrated BxPc-3 cells in the experimental group was far less than in control (P < 0.05). In vivo, the tumor size and weight in the experimental group were significantly lower than those in control (P < 0.05). CONCLUSION: Our data demonstrate that antisense HIF-1alpha inhibits expressions of survivin and beta1 integrin, enhancing apoptosis in human pancreatic cancer cells and restraining the progression and metastasis of pancreatic cancer. Therefore, HIF-1alpha may play a very important role in progression, metastasis, and chemosensitivity of human pancreatic cancer. Blocking HIF-1alpha in pancreatic cancer cells may offer an avenue for gene therapy.
OBJECTIVES: The aim of the study was to observe the effect of antisense hypoxia-inducible factor 1alpha (HIF-1alpha) on progression, metastasis, and chemosensitivity of pancreatic cancer. METHODS: BxPc-3 cells transfected with antisense HIF-1alpha plasmid were exposed to 0.5% O2 for 4 hours. Expressions of HIF-1alpha, survivin, and beta1 integrin were detected by reverse transcriptase -polymerase chain reaction and Western blotting. Growth inhibition rates and apoptosis rates of BxPc-3 cells under different dosages of chemotherapy agents (5-fluorouracil, doxorubicin, and gemcitabine) were measured by MTT colorimetric assay and flow cytometry. The migration of BxPc-3 cells was assayed using transwell cell culture chambers. Subcutaneous transplantation of BxPc-3 cells in nude mice for 8 weeks was to assess progression and metastasis of pancreatic cancer. RESULTS: Expression of HIF-1alpha was obviously down-regulated, and at the same time, survivin and beta1-integrin expressions were markedly down-regulated in the experimental group (P < 0.05). Higher dosages (100, 200, and 400 mg/L of 5-fluorouracil; 0.05, 0.075, and 0.1 mg/L of doxorubicin; and 10(-9), 10(-8), and 10(-7) mol/L of gemcitabine) caused a greater increase of inhibition in the experimental group than in control (P < 0.05). The number of migrated BxPc-3 cells in the experimental group was far less than in control (P < 0.05). In vivo, the tumor size and weight in the experimental group were significantly lower than those in control (P < 0.05). CONCLUSION: Our data demonstrate that antisense HIF-1alpha inhibits expressions of survivin and beta1 integrin, enhancing apoptosis in humanpancreatic cancer cells and restraining the progression and metastasis of pancreatic cancer. Therefore, HIF-1alpha may play a very important role in progression, metastasis, and chemosensitivity of humanpancreatic cancer. Blocking HIF-1alpha in pancreatic cancer cells may offer an avenue for gene therapy.
Authors: Takaaki Imamura; Hirotoshi Kikuchi; Maria-Teresa Herraiz; Do-Youn Park; Yusuke Mizukami; Mari Mino-Kenduson; Maureen P Lynch; Bo R Rueda; Yair Benita; Ramnik J Xavier; Daniel C Chung Journal: Int J Cancer Date: 2009-02-15 Impact factor: 7.396