Anti-tumor angiogenesis therapy, targeting the suppression of blood vessel growth in tumors, presents a potent approach in the battle against cancer. Traditional therapies have primarily concentrated on single-target ...Anti-tumor angiogenesis therapy, targeting the suppression of blood vessel growth in tumors, presents a potent approach in the battle against cancer. Traditional therapies have primarily concentrated on single-target techniques, with a specific emphasis on targeting the vascular endothelial growth factor, but have not reached ideal therapeutic efficacy. In response to this issue, our study introduced a novel nanoparticle system known as CS-siRNA/PEITC&L-cRGD NPs. These chitosan-based nanoparticles have been recognized for their excellent biocompatibility and ability to deliver genes. To enhance their targeted delivery capability, they were combined with a cyclic RGD peptide (cRGD). Targeted co-delivery of gene and chemotherapeutic agents was achieved through the use of a negatively charged lipid shell and cRGD, which possesses high affinity for integrin αvβ3 overexpressed in tumor cells and neovasculature. In this multifaceted approach, co-delivery of VEGF siRNA and phenethyl isothiocyanate (PEITC) was employed to target both tumor vascular endothelial cells and tumor cells simultaneously. The co-delivery of VEGF siRNA and PEITC could achieve precise silencing of VEGF, inhibit the accumulation of HIF-1α under hypoxic conditions, and induce apoptosis in tumor cells. In summary, we have successfully developed a nanoparticle delivery platform that utilizes a dual mechanism of action of anti-tumor angiogenesis and pro-tumor apoptosis, which provides a robust and potent strategy for the delivery of anti-cancer therapeutics.展开更多
Objective:To investigate the vasculogenic mimicry formation induced by hypoxia in Ⅱ-Ⅲ human glioma cell and the effect of alphastatin peptide suppressing the hypoxia-induced vasculogenic mimicry formation and the me...Objective:To investigate the vasculogenic mimicry formation induced by hypoxia in Ⅱ-Ⅲ human glioma cell and the effect of alphastatin peptide suppressing the hypoxia-induced vasculogenic mimicry formation and the mechanism.Methods:MTT,Transwell and three-dimentional culture were used to detect the proliferation,migration and tubule formation of SHG44.The expression of vascular endothelial growth factor-α(VEGF-α),erythropoietin-producing hepatocellular carcinoma-A2 (EphA2) and matrix metalloproteinases 2 (MMP2) was detected by RT-PCR and Western blotting analysis.Results:The OD 490 in hypoxia group was 0.60±0.06 and in control group was 0.46±0.05.The number of cell migration was 178.71±18.81 in hypoxia group and 85.86±17.92 in control group.The tubule formation was 56.80±12.21 in hypoxia group and 4.20±2.62 in control group.The proliferation,migration and tubule formation in hypoxia group were significantly higher than that in control group.The expression of VEGF-α,EphA2 and MMP2 was upregulated in hypoxia.When various concentrations of alphastatin (100,1 000,10 000 nmol/L) were added to hypoxia group,the numbers of cell migration were 142.57±12.12,92.71±17.68,30.00±7.72 and the tubule formation were 47.71±10.58,18.86±8.40,8.43±5.62.The cell migration and tubule formation were significantly suppressed by alphastatin in a dose-dependent manner.In alphastatin group,the phosphorylation of EphA2 protein (P=0.037,F=4.629) and activation of MMP2 protein (P=0.005,F=9.331) were significantly suppressed but there was no change in VEGF-α protein.Conclusion:Ⅱ-Ⅲ human glioma cell is able to form vasculogenic mimicry induced by hypoxia and alphastatin peptide can suppress the hypoxia-induced vasculogenic mimicry.VEGF-α induced EphA2 phospharilation and MMP2 activation maybe the key pathway to form vasculogenic mimicry.展开更多
Hypoxia affects the advancement,metastasis,and metabolism of breast cancer(BC).The circular RNA ribonuclease P RNA component H1(circRPPH1)(has_circ_0000515)is implicated in tumor progression.Nevertheless,the regulator...Hypoxia affects the advancement,metastasis,and metabolism of breast cancer(BC).The circular RNA ribonuclease P RNA component H1(circRPPH1)(has_circ_0000515)is implicated in tumor progression.Nevertheless,the regulatory mechanism related to circRPPH1 in hypoxia-mediated triple-negative breast cancer(TNBC)progression is indistinct.The expression levels of circRPPH1,miR-1296-5p,tripartite motif-containing 14(TRIM14)mRNA in tissue samples and cells were examined through quantitative real-time polymerase chain reaction(qRT-PCR).Cell viability,migration,and invasion were determined with Cell Counting Kit-8(CCK-8)or transwell assays.The levels of glucose consumption and lactate production were assessed via the Glucose Assay Kit or Lactate Assay Kit.The protein levels of TRIM14,Glucose Transporter GLUT1(GLUT1),and lactic dehydrogenase A(LDHA)were detected by western blot analysis.The targeting relationship between circRPPH1 or TRIM14 and miR-1296-5p was verified with dual-luciferase reporter assay.The role of circRPPH1 was confirmed via xenograft assay.We verified that circRPPH1 and TRIM14 expression were increased while miR-1296-5p expression was decreased in BC tissues and hypoxia-cultured TNBC cells.Functionally,circRPPH1 silencing reversed the promoting effect of hypoxia on viability,migration,invasion,and glycolysis of TNBC cells.CircRPPH1 knockdown repressed decreased TNBC cell growth in vivo.Mechanistically,circRPPH1 sponged miR-1296-5p to modulate TRIM14 expression.Also,miR-1296-5p silencing restored circRPPH1 inhibition-mediated influence on the viability,migration,invasion,and glycolysis of hypoxia-treated TNBC cells.TRIM14 elevation overturned the inhibitory impact of miR-1296-5p mimic on viability,migration,invasion,and glycolysis of hypoxia-cultured TNBC cells.In conclusion,hypoxia-induced circRPPH1 fostered TNBC progression through regulation of the miR-1296-5p/TRIM14 axis,indicating that circRPPH1 was a promising target for TNBC treatment.展开更多
Objective:To investigate the expression of HSP90 and HIF-1αin human colorectal cancer tissue,the influence of HSP90 and HIF-1αon human colorectal cancer biological behavior and their related factors.Methods:The expr...Objective:To investigate the expression of HSP90 and HIF-1αin human colorectal cancer tissue,the influence of HSP90 and HIF-1αon human colorectal cancer biological behavior and their related factors.Methods:The expression of HSP90 and HIF-1 a protein in human colorectal cancer as well as normal tissue were detected by imnmnohistochemical method.Results:The positive expression rates of HSP90 and HIF-1αprotein in normal human colorectal tissue as well as colorectal cancer tissue were 30%vs.63.0%,15.0%vs.71.7%,respectively.There were significant difference(P=0.035 and P=0.005 respectively).The expression of HSP90 was significantly correlated with the differentiation,Dukes stages and lymph node metastasis(P<0.05),while the expression of HIF-1 a was significantly correlated with the Dukes stages and lymph node metastasis(P<0.05).Association analysis showed that the expression of HSF90 protein was significantly correlated with that of HIF-1αprotein(P<0.01).Conclusions:The expression of HSP90 and HIF—1αprotein may be related to the development,metastasis and invasion of human colorectal cancer,and their synergistic effects may participate in the development of the colorectal carcinoma.展开更多
This study demonstrated that damage to the cerebral microvasculature, the formation of microthrombi and swelling of vascular endothelial cells occur early and peak 12 hours after injury in a rat model of diffuse axona...This study demonstrated that damage to the cerebral microvasculature, the formation of microthrombi and swelling of vascular endothelial cells occur early and peak 12 hours after injury in a rat model of diffuse axonal injury. Moreover, these pathological changes were most evident in the cerebral cortex. Cerebral microcirculatory dysfunction peaked later and had a shorter duration than axonal injury. In addition, the radioactive imaging agent, 99Tcm-4, 9-diaza-2, 3, 10, 10- tetramethyldodecan-2, 11 -dione dioxime, was used to visualize the dynamic changes that occur in tissue with cerebral hypoxia. The results demonstrated that cerebral hypoxia occurs at an early stage in diffuse axonal injury. Cerebral hypoxia was evident 12 hours after injury and declined slightly 24 hours after injury, but was significantly higher than in the control group. The pathological changes that underpin microcirculatory dysfunction did not occur at the same time as axonal injury, but did occur simultaneously with neuronal injury. Cerebral hypoxia plays a key role in promoting the secondary brain injury that occurs after diffuse axonal injury.展开更多
Calcium-dependent protein kinases(CDPKs/CPKs)are key regulators of plant stress signaling that translate calcium signals into cellular responses by phosphorylating diverse substrate proteins.However,the molecular mech...Calcium-dependent protein kinases(CDPKs/CPKs)are key regulators of plant stress signaling that translate calcium signals into cellular responses by phosphorylating diverse substrate proteins.However,the molecular mechanism by which plant cells relay calcium signals in response to hypoxia remains elusive.Here,we show that one member of the CDPK family in Arabidopsis thaliana,CPK12,is rapidly activated during hypoxia through calcium-dependent phosphorylation of its Ser-186 residue.Phosphorylated CPK12 shuttles from the cytoplasm to the nucleus,where it interacts with and phosphorylates the group Ⅶ ethylene-responsive transcription factors(ERF-Ⅶ)that are core regulators of plant hypoxia sensing,to enhance their stabilities.Consistently,CPK12 knockdown lines show attenuated tolerance of hypoxia,whereas transgenic plants overexpressing CPK12 display improved hypoxia tolerance.Nonethelss,loss of function of five ERF-Ⅶ proteins in an erf-vii pentuple mutant could partially suppress the enhanced hypoxia-tolerance phenotype of CPK12-overexpressing lines.Moreover,we also discovered that phosphatidic acid and 14-3-3κ protein serve as positive and negative modulators of the CPK12 cytoplasm-to-nucleus translocation,respectively.Taken together,these findings uncover a CPK12-ERF-Ⅶ regulatory module that is key to transducing calcium signals from the cytoplasm into the nucleus to potentiate hypoxia sensing in plants.展开更多
基金supported by Guangdong Basic and Applied Basic Research Foundation(2023A1515010969)Natural Science Foundation of Top Talent of SZTU(GDRC202305).
文摘Anti-tumor angiogenesis therapy, targeting the suppression of blood vessel growth in tumors, presents a potent approach in the battle against cancer. Traditional therapies have primarily concentrated on single-target techniques, with a specific emphasis on targeting the vascular endothelial growth factor, but have not reached ideal therapeutic efficacy. In response to this issue, our study introduced a novel nanoparticle system known as CS-siRNA/PEITC&L-cRGD NPs. These chitosan-based nanoparticles have been recognized for their excellent biocompatibility and ability to deliver genes. To enhance their targeted delivery capability, they were combined with a cyclic RGD peptide (cRGD). Targeted co-delivery of gene and chemotherapeutic agents was achieved through the use of a negatively charged lipid shell and cRGD, which possesses high affinity for integrin αvβ3 overexpressed in tumor cells and neovasculature. In this multifaceted approach, co-delivery of VEGF siRNA and phenethyl isothiocyanate (PEITC) was employed to target both tumor vascular endothelial cells and tumor cells simultaneously. The co-delivery of VEGF siRNA and PEITC could achieve precise silencing of VEGF, inhibit the accumulation of HIF-1α under hypoxic conditions, and induce apoptosis in tumor cells. In summary, we have successfully developed a nanoparticle delivery platform that utilizes a dual mechanism of action of anti-tumor angiogenesis and pro-tumor apoptosis, which provides a robust and potent strategy for the delivery of anti-cancer therapeutics.
基金Supported in Part by a Grant from the National Nature Science Foundation of China(No.30672126)
文摘Objective:To investigate the vasculogenic mimicry formation induced by hypoxia in Ⅱ-Ⅲ human glioma cell and the effect of alphastatin peptide suppressing the hypoxia-induced vasculogenic mimicry formation and the mechanism.Methods:MTT,Transwell and three-dimentional culture were used to detect the proliferation,migration and tubule formation of SHG44.The expression of vascular endothelial growth factor-α(VEGF-α),erythropoietin-producing hepatocellular carcinoma-A2 (EphA2) and matrix metalloproteinases 2 (MMP2) was detected by RT-PCR and Western blotting analysis.Results:The OD 490 in hypoxia group was 0.60±0.06 and in control group was 0.46±0.05.The number of cell migration was 178.71±18.81 in hypoxia group and 85.86±17.92 in control group.The tubule formation was 56.80±12.21 in hypoxia group and 4.20±2.62 in control group.The proliferation,migration and tubule formation in hypoxia group were significantly higher than that in control group.The expression of VEGF-α,EphA2 and MMP2 was upregulated in hypoxia.When various concentrations of alphastatin (100,1 000,10 000 nmol/L) were added to hypoxia group,the numbers of cell migration were 142.57±12.12,92.71±17.68,30.00±7.72 and the tubule formation were 47.71±10.58,18.86±8.40,8.43±5.62.The cell migration and tubule formation were significantly suppressed by alphastatin in a dose-dependent manner.In alphastatin group,the phosphorylation of EphA2 protein (P=0.037,F=4.629) and activation of MMP2 protein (P=0.005,F=9.331) were significantly suppressed but there was no change in VEGF-α protein.Conclusion:Ⅱ-Ⅲ human glioma cell is able to form vasculogenic mimicry induced by hypoxia and alphastatin peptide can suppress the hypoxia-induced vasculogenic mimicry.VEGF-α induced EphA2 phospharilation and MMP2 activation maybe the key pathway to form vasculogenic mimicry.
基金was funded by the Xinjiang Uygur Autonomous Region Natural Science Foundation(No.2017D01C379).
文摘Hypoxia affects the advancement,metastasis,and metabolism of breast cancer(BC).The circular RNA ribonuclease P RNA component H1(circRPPH1)(has_circ_0000515)is implicated in tumor progression.Nevertheless,the regulatory mechanism related to circRPPH1 in hypoxia-mediated triple-negative breast cancer(TNBC)progression is indistinct.The expression levels of circRPPH1,miR-1296-5p,tripartite motif-containing 14(TRIM14)mRNA in tissue samples and cells were examined through quantitative real-time polymerase chain reaction(qRT-PCR).Cell viability,migration,and invasion were determined with Cell Counting Kit-8(CCK-8)or transwell assays.The levels of glucose consumption and lactate production were assessed via the Glucose Assay Kit or Lactate Assay Kit.The protein levels of TRIM14,Glucose Transporter GLUT1(GLUT1),and lactic dehydrogenase A(LDHA)were detected by western blot analysis.The targeting relationship between circRPPH1 or TRIM14 and miR-1296-5p was verified with dual-luciferase reporter assay.The role of circRPPH1 was confirmed via xenograft assay.We verified that circRPPH1 and TRIM14 expression were increased while miR-1296-5p expression was decreased in BC tissues and hypoxia-cultured TNBC cells.Functionally,circRPPH1 silencing reversed the promoting effect of hypoxia on viability,migration,invasion,and glycolysis of TNBC cells.CircRPPH1 knockdown repressed decreased TNBC cell growth in vivo.Mechanistically,circRPPH1 sponged miR-1296-5p to modulate TRIM14 expression.Also,miR-1296-5p silencing restored circRPPH1 inhibition-mediated influence on the viability,migration,invasion,and glycolysis of hypoxia-treated TNBC cells.TRIM14 elevation overturned the inhibitory impact of miR-1296-5p mimic on viability,migration,invasion,and glycolysis of hypoxia-cultured TNBC cells.In conclusion,hypoxia-induced circRPPH1 fostered TNBC progression through regulation of the miR-1296-5p/TRIM14 axis,indicating that circRPPH1 was a promising target for TNBC treatment.
文摘目的:探讨新生大鼠缺氧/缺血性脑损伤时皮层神经元缺氧诱导因子-1α(HIF-1α)的表达及其与凋亡相关基因P53、Bcl-2的关系。方法:新生7日龄SD大鼠随机分为假手术组、单纯缺氧组和缺氧/缺血组,在缺氧或缺氧/缺血后3、6、12、24、72 h断头取脑,HE染色观察脑组织病理学改变,免疫组织化学方法检测皮层神经元HIF-1α、P53及Bcl-2表达情况。结果:单纯缺氧组和缺氧/缺血组大鼠脑组织HE染色显示:脑皮层神经元均有不同程度的损伤,于24 h时损伤最重,细胞溶解缺失明显;皮层HIF-1α于缺氧/缺血后3 h表达升高,12 h达高峰,之后呈下降趋势;皮层P53表达高峰较HIF-1α推后,24 h达高峰,之后呈下降趋势;Bcl-2表达规律与HIF-1α一致。假手术组P53/Bcl-2值约等于1;单纯缺氧组和缺氧/缺血组3、6、12 h 3个时间点上P53/Bcl-2值小于1,24、72 h 2个时间点上P53/Bcl-2值大于1。结论:在新生大鼠缺氧/缺血性脑损伤时,HIF-1α参与了对P53及Bcl-2的调控,在缺氧损伤早期可能具有抗凋亡作用。
基金supported by Shaanxi province health department key funds:sx201227273
文摘Objective:To investigate the expression of HSP90 and HIF-1αin human colorectal cancer tissue,the influence of HSP90 and HIF-1αon human colorectal cancer biological behavior and their related factors.Methods:The expression of HSP90 and HIF-1 a protein in human colorectal cancer as well as normal tissue were detected by imnmnohistochemical method.Results:The positive expression rates of HSP90 and HIF-1αprotein in normal human colorectal tissue as well as colorectal cancer tissue were 30%vs.63.0%,15.0%vs.71.7%,respectively.There were significant difference(P=0.035 and P=0.005 respectively).The expression of HSP90 was significantly correlated with the differentiation,Dukes stages and lymph node metastasis(P<0.05),while the expression of HIF-1 a was significantly correlated with the Dukes stages and lymph node metastasis(P<0.05).Association analysis showed that the expression of HSF90 protein was significantly correlated with that of HIF-1αprotein(P<0.01).Conclusions:The expression of HSP90 and HIF—1αprotein may be related to the development,metastasis and invasion of human colorectal cancer,and their synergistic effects may participate in the development of the colorectal carcinoma.
基金the National Natural Science Foundationof China, No. 30471774the Program for New Century Excellent Talents in University, Ministry of Education,China, No. NCET-05-0831
文摘This study demonstrated that damage to the cerebral microvasculature, the formation of microthrombi and swelling of vascular endothelial cells occur early and peak 12 hours after injury in a rat model of diffuse axonal injury. Moreover, these pathological changes were most evident in the cerebral cortex. Cerebral microcirculatory dysfunction peaked later and had a shorter duration than axonal injury. In addition, the radioactive imaging agent, 99Tcm-4, 9-diaza-2, 3, 10, 10- tetramethyldodecan-2, 11 -dione dioxime, was used to visualize the dynamic changes that occur in tissue with cerebral hypoxia. The results demonstrated that cerebral hypoxia occurs at an early stage in diffuse axonal injury. Cerebral hypoxia was evident 12 hours after injury and declined slightly 24 hours after injury, but was significantly higher than in the control group. The pathological changes that underpin microcirculatory dysfunction did not occur at the same time as axonal injury, but did occur simultaneously with neuronal injury. Cerebral hypoxia plays a key role in promoting the secondary brain injury that occurs after diffuse axonal injury.
基金supported by the National Natural Science Foundation of China(Projects 31725004,U22A20458)the Key Realm Research and Development Program of Guangdong Province(Project 2020B0202090001)the Natural Science Foundation of Guangdong Province(Project 2023A1515012038).
文摘Calcium-dependent protein kinases(CDPKs/CPKs)are key regulators of plant stress signaling that translate calcium signals into cellular responses by phosphorylating diverse substrate proteins.However,the molecular mechanism by which plant cells relay calcium signals in response to hypoxia remains elusive.Here,we show that one member of the CDPK family in Arabidopsis thaliana,CPK12,is rapidly activated during hypoxia through calcium-dependent phosphorylation of its Ser-186 residue.Phosphorylated CPK12 shuttles from the cytoplasm to the nucleus,where it interacts with and phosphorylates the group Ⅶ ethylene-responsive transcription factors(ERF-Ⅶ)that are core regulators of plant hypoxia sensing,to enhance their stabilities.Consistently,CPK12 knockdown lines show attenuated tolerance of hypoxia,whereas transgenic plants overexpressing CPK12 display improved hypoxia tolerance.Nonethelss,loss of function of five ERF-Ⅶ proteins in an erf-vii pentuple mutant could partially suppress the enhanced hypoxia-tolerance phenotype of CPK12-overexpressing lines.Moreover,we also discovered that phosphatidic acid and 14-3-3κ protein serve as positive and negative modulators of the CPK12 cytoplasm-to-nucleus translocation,respectively.Taken together,these findings uncover a CPK12-ERF-Ⅶ regulatory module that is key to transducing calcium signals from the cytoplasm into the nucleus to potentiate hypoxia sensing in plants.
