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Objective • To study the effect of inhibitor of differentiation 1 (ID1) on ocular neovascularization. Methods • The oxygen-induced retinal neovascularization (OIR), laser-induced choroidal neovascularization (CNV) and over-expression of vascular endothelial growth factor (VEGF) (Rho-VEGF) transgenic mice were established. The localization and mRNA level of ID1 in retina of OIR mice and Rho-VEGF transgenic mice were determined by immunofluorescence staining and quantitative real-time PCR. Mice deficient in ID1 (ID1-/-) were used to induce retinal neovascularization in accordance with the above three models, and to compare the changes of ID1 on the number of retinal, subretinal and choroidal neovascularization areas. In order to explore the role ID1 in neovascularization, the numbers and areas of retinal, subretinal and choroidal neovascularization in the mice models with or without ID1 deficiency were compared. Its effect on the related factors, i.e. hypoxia-inducible factor-1α (HIF-1α), VEGF and vascular endothelial growth factor receptor 1/2 (VEGFR1/2) were also observed. Results • Mice deficient in ID1 showed a significant reduction in the area of neovascularization in these three models(P<0.05). Mice lacking ID1 showed reduced levels of HIF-1α, VEGF and VEGFR 1. Conclusion • ID1 promotes the expression of HIF-1α, VEGF and VEGFR1 in the retina and choroidal neovascularization during hypoxia and oxidative injury.
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Objective · To study the effect of inhibitor of differentiation 1 (ID1) on ocular neovascularization. Methods · The oxygen-induced retinal neovascularization (OIR), laser-induced choroidal neovascularization (CNV) and over-expression of vascular endothelial growth factor (VEGF) (Rho-VEGF) transgenic mice were established. The localization and mRNA level of ID1 in retina of OIR mice and Rho-VEGF transgenic mice were determined by immunofluorescence staining and quantitative real-time PCR. Mice deficient in ID1 (ID1-/-) were used to induce retinal neovascularization in accordance with the above three models, and to compare the changes of ID1 on the number of retinal, subretinal and choroidal neovascularization areas. In order to explore the role ID1 in neovascularization, the numbers and areas of retinal, subretinal and choroidal neovascularization in the mice models with or without ID1 deficiency were compared. Its effect on the related factors, i.e. hypoxia-inducible factor-1α (HIF-1α), VEGF and vascular endothelial growth factor receptor 1/2 (VEGFR1/2) were also observed. Results · Mice deficient in ID1 showed a significant reduction in the area of neovascularization in these three models (P＜0.05). Mice lacking ID1 showed reduced levels of HIF-1α, VEGF and VEGFR 1. Conclusion · ID1 promotes the expression of HIF-1α, VEGF and VEGFR1 in the retina and choroidal neovascularization during hypoxia and oxidative injury.
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@# Objective: To investigate whether inhibitor of differentiation 1 gene (Id1) and Id3 gene can synergistically promote epithelial-mesenchymal transition (EMT), invasion and migration of colon cancer SW620 cells and to explore its underlying mechanisms. Methods: The SW620 cell strain with Idl or Id3 gene knockdown and the SW620 cell strain with Id1/Id3 gene double-knockdown were constructed by lentiviral vectors transfection. The SW620 cells were divided into four groups, which included SW620-Sh-Id1 group (transfected with shRNA-Id1), SW620-Sh-Id3 group (transfected with shRNA-Id3), SW620-Sh-Id1-Id3 group (transfected with shRNAId1 plus shRNA-Id3) and SW620-NC group (transfected with negative lentivirus). The efficiency of knockdown was detected by Realtime qPCR and Western blotting. The influence of stable knockdown of Idl or Id3 on cell morphological change was observed under a microscope. The changes of migration and invasion abilities of the SW620 cells were determined by wound healing assay and Transwell assay. EMT, invasion and migration related proteins were measured by Western blotting. Results: The SW620 cell strains with Idl and/or Id3 gene knockdown were successfully constructed. Idl and Id3 knockdown induced the epithelial-like to the mesenchymanl-like transformation of SW620 cells. (1) Compared with the control group, the invasion and migration abilities of the SW620 cells were significantly decreased in the SW620-Sh-Id1 group and SW620-Sh-Id3 group (all P<0.05). (2) Meanwhile, the invasion and migration abilities in the SW620-Sh-Id1-Id3 group were obviously weaker than the SW620-Sh-Id1 group and SW620-Sh-Id3 group (all P<0.05). (3) Compared with the control group, the SW620-Sh-Id1 group and SW620-Sh-Id3 group had a reduction in the protein expressions of βcatenin, snail1 and MMP2, and an increase in the protein expressions of E-cadherin and TIMP2 (all P<0.05). (4) Compared with the SW620-Sh-Id1 group and SW620-Sh-Id3 group , the protein expressions of β-catenin, snail1 and MMP2 were reduced, and the protein expressions of E-cadherin and TIMP2 were increased in the SW620-Sh-Id1-Id3 group (all P<0.05). Conclusion: Id1 and Id3 could synergistically influence invasion and migration of SW620 cells, possibly through inducing EMT.
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Objective To investigate the expressions and clinical significance of Id1 in diffuse large B‐cell Lymphomas (DL‐BCL) tissue and Id1 gene in bone warrow cell .Methods Forty cases of DLBCL(observation group) and 25 cases of reactive lymph‐oid hyperplasia (control group) were included in this study which were admitted by our hospital from October 2011 to October 2014 .The expression of Id1 proteins in DLBCL and reactive lymphoid hyperplasia were detected by imunohistochemical technique . The expression of Id1 genes in all patients′marrow cells was detected by reverse‐transcriptase polymerase chain reaction .The data were collected and analyzed by designed person .Results In 40 DLBCLs ,the positive rate of Id1 were 75 .00% (30/40) ,which was higher than in RHs 32 .00% (8/25) ,with statistic difference(P= 0 .001) .Id1 protein was not correlated with sex and age(P>0 .05) ,but was correlated with clinical stage ,LDH level and extranodal infiltration(P<0 .05) .The expression of Id1 genes in mar‐row cells in DLBCL was higher than in RH (Id1mRNA level 2 .80 ± 0 .87 vs .1 .37 ± 0 .51 ,P<0 .05) ,and also correlated with clini‐cal stage ,LDH level and extranodal infiltration(P<0 .05) .Conclusion The expression of Id1 proteins and genes are much higher in DLBCL tissue and marrow and probably related to the prognosis of DLBCL .This discovery would contribute to predicting prognosis of DLBCL ,w hich also could be a therapeutic target of DLBCL in the future .
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Objective: To explore the role and the possible mechanism of inhibitor of differentiation 1 (Id1) in angiogenesis of colon cancer. Methods: After transfection with Id1 over-expression vector plasmid, the expression levels of hypoxia-inducible factor-1α (HIF-1α) and vascular endothelial growth factor (VEGF) mRNAs and proteins in HT-29 cells were detected by real-time fluorescent quantitative PCR and Western blotting, respectively. The ability of tube formation of human umbilical vein endothelial cells (HUVECs) after culture with supernatant of HT-29 cells with Id1 overexpression for 24 h was observed by tube formation experiment. The tumorigenesis of HT-29 cells with Id1 over-expression in nude mice was observed, and the microvessel density (MVD) of transplanted tumor tissues was detected by immunohistochemistry, . Results: The expression levels of HIF-1α and VEGF mRNAs and proteins in HT-29 cells with Id1 over-expression were higher than those in the HT-29 cells transfected with empty vector (negative control) and the HT-29 cells without any transfection (blank control) (all P < 0.05). The number of tube formation of HUVECs after culture with supernatant of HT-29 cells with Id1 over-expression was higher than those of the negative control and the blank control groups (both P < 0.05). The ability of tmorigenesis and the number of MVD in Id1 overexpression group were both higher than those in the negative control and the blank control groups (all P < 0.05). Conclusion: Id1 may promote the angiogenesis of colon cancer by regulating the expressions of HIF-1α and VEGF.