AIM: To investigate the effect of integrin-linked kinase(ILK) on proliferation, metastasis, and invasion of the colorectal cancer cell line SW480. METHODS: In this study, the colorectal cancer cell line SW480 was stab...AIM: To investigate the effect of integrin-linked kinase(ILK) on proliferation, metastasis, and invasion of the colorectal cancer cell line SW480. METHODS: In this study, the colorectal cancer cell line SW480 was stably transfected with ILK plasmids, and small interfering RNA(si RNA) was used to knockdown expression of nuclear factor(NF)-κB/p65. Methylthiazole tetrazolium(MTT) assay was performed to measure proliferation, and the wound healing migration assay and matrigel invasion assay were used to test the metastasis and invasion ability of SW480 cells. To explore the epithelial-mesenchymal transition(EMT) process, embryonic development, and the invasion and metastasis of tumors, the protein level of E-cadherin, vimentin, snail, and slug was detected by western blot. Immunofluorescence was also used to detect E-cadherin expression. Western blot was used to determine the level of phosphorylated-inhibitor of kappa B(IκB)a, inhibitor of gamma B(IγB)a, and nuclear factor kappa B(NF-κB) expressions and toexplore the ILK signaling pathway. RESULTS: Western blot results revealed that ILK expression significantly increased when ILK was overexpressed in SW480 cells(P < 0.05). Proliferation, metastasis, and invasion ability were improved in the vector-ILK group compared to the vector group(P < 0.05). Immunofluorescence results revealed that E-cadherin fluorescence intensity decreased after ILK was overexpressed(P < 0.05). Western blot results revealed that the protein expression of E-cadherin was reduced, while vimentin, snail, and slug were upregulated when ILK was overexpressed in SW480 cells(P < 0.05). In order to determine the role of the NF-κB signaling pathway in ILK overexpression promoted EMT occurrence, we overexpressed ILK in SW480 cells and found that levels of NF-κB/p65 and cytoplasmic phosphorylated-IκBa were increased and that cytoplasmic IкBa levels were decreased compared to the control group(P < 0.05). Furthermore, NF-κB/p65 knockout revealed that E-cadherin was increased in the overexpressed ILK group. CONCLUSION: ILK overexpression improved the proliferation, metastasis, and invasion ability of SW480 cells, and this effect may be mediated by the NF-κB signaling pathway.展开更多
AIM:To investigate the effects of integrin-linked kinase(ILK)on gastric cancer cells both in vitro and in vivo.METHODS:ILK small interfering RNA(siRNA)was transfected into human gastric cancer BGC-823 cells and ILK ex...AIM:To investigate the effects of integrin-linked kinase(ILK)on gastric cancer cells both in vitro and in vivo.METHODS:ILK small interfering RNA(siRNA)was transfected into human gastric cancer BGC-823 cells and ILK expression was monitored by real-time quantitative polymerase chain reaction,Western blotting analysis and immunocytochemistry.Cell attachment,proliferation,invasion,microfilament dynamics and the secretion of vascular endothelial growth factor(VEGF) were also measured.Gastric cancer cells treated with ILK siRNA were subcutaneously transplanted into nude mice and tumor growth was assessed.RESULTS:Both ILK mRNA and protein levels were significantly down-regulated by ILK siRNA in human gastric cancer cells.This significantly inhibited cell attachment,proliferation and invasion.The knockdown ofILK also disturbed F-actin assembly and reduced VEGF secretion in conditioned medium by 40%(P<0.05).Four weeks after injection of ILK siRNA-transfected gastric cancer cells into nude mice,tumor volume and weight were significantly reduced compared with that of tumors induced by cells treated with non-silencing siRNA or by untreated cells(P<0.05).CONCLUSION:Targeting ILK with siRNA suppresses the growth of gastric cancer cells both in vitro and in vivo.ILK plays an important role in gastric cancer progression.展开更多
Background Integrin-linked kinase （ILK） dysregulation is involved in the progression of diabetic nephropathy （DN）. The aim of this study was to investigate the effects of angiotensin II receptor blocker （ARB）, i...Background Integrin-linked kinase （ILK） dysregulation is involved in the progression of diabetic nephropathy （DN）. The aim of this study was to investigate the effects of angiotensin II receptor blocker （ARB）, irbesartan, on ILK expression and podocyte injury in DN.展开更多
基金Supported by the National Natural Science Foundation of China(Beijing,China,grant No’s.30770971,81172470,81070362 and 81372629)
文摘AIM: To investigate the effect of integrin-linked kinase(ILK) on proliferation, metastasis, and invasion of the colorectal cancer cell line SW480. METHODS: In this study, the colorectal cancer cell line SW480 was stably transfected with ILK plasmids, and small interfering RNA(si RNA) was used to knockdown expression of nuclear factor(NF)-κB/p65. Methylthiazole tetrazolium(MTT) assay was performed to measure proliferation, and the wound healing migration assay and matrigel invasion assay were used to test the metastasis and invasion ability of SW480 cells. To explore the epithelial-mesenchymal transition(EMT) process, embryonic development, and the invasion and metastasis of tumors, the protein level of E-cadherin, vimentin, snail, and slug was detected by western blot. Immunofluorescence was also used to detect E-cadherin expression. Western blot was used to determine the level of phosphorylated-inhibitor of kappa B(IκB)a, inhibitor of gamma B(IγB)a, and nuclear factor kappa B(NF-κB) expressions and toexplore the ILK signaling pathway. RESULTS: Western blot results revealed that ILK expression significantly increased when ILK was overexpressed in SW480 cells(P < 0.05). Proliferation, metastasis, and invasion ability were improved in the vector-ILK group compared to the vector group(P < 0.05). Immunofluorescence results revealed that E-cadherin fluorescence intensity decreased after ILK was overexpressed(P < 0.05). Western blot results revealed that the protein expression of E-cadherin was reduced, while vimentin, snail, and slug were upregulated when ILK was overexpressed in SW480 cells(P < 0.05). In order to determine the role of the NF-κB signaling pathway in ILK overexpression promoted EMT occurrence, we overexpressed ILK in SW480 cells and found that levels of NF-κB/p65 and cytoplasmic phosphorylated-IκBa were increased and that cytoplasmic IкBa levels were decreased compared to the control group(P < 0.05). Furthermore, NF-κB/p65 knockout revealed that E-cadherin was increased in the overexpressed ILK group. CONCLUSION: ILK overexpression improved the proliferation, metastasis, and invasion ability of SW480 cells, and this effect may be mediated by the NF-κB signaling pathway.
基金Supported by The grants from the Department of Anesthesiology and Intensive Care of Changhai Hospital,Shanghai,China
文摘AIM:To investigate the effects of integrin-linked kinase(ILK)on gastric cancer cells both in vitro and in vivo.METHODS:ILK small interfering RNA(siRNA)was transfected into human gastric cancer BGC-823 cells and ILK expression was monitored by real-time quantitative polymerase chain reaction,Western blotting analysis and immunocytochemistry.Cell attachment,proliferation,invasion,microfilament dynamics and the secretion of vascular endothelial growth factor(VEGF) were also measured.Gastric cancer cells treated with ILK siRNA were subcutaneously transplanted into nude mice and tumor growth was assessed.RESULTS:Both ILK mRNA and protein levels were significantly down-regulated by ILK siRNA in human gastric cancer cells.This significantly inhibited cell attachment,proliferation and invasion.The knockdown ofILK also disturbed F-actin assembly and reduced VEGF secretion in conditioned medium by 40%(P<0.05).Four weeks after injection of ILK siRNA-transfected gastric cancer cells into nude mice,tumor volume and weight were significantly reduced compared with that of tumors induced by cells treated with non-silencing siRNA or by untreated cells(P<0.05).CONCLUSION:Targeting ILK with siRNA suppresses the growth of gastric cancer cells both in vitro and in vivo.ILK plays an important role in gastric cancer progression.
基金This study was supported by a grant from the National Natural Science Foundation of China （No. 30870953） to Prof. LIU Bi-cheng as PI.
文摘Background Integrin-linked kinase （ILK） dysregulation is involved in the progression of diabetic nephropathy （DN）. The aim of this study was to investigate the effects of angiotensin II receptor blocker （ARB）, irbesartan, on ILK expression and podocyte injury in DN.
