宫颈鳞状上皮癌变过程中缺氧诱导因子1α的表达及意义

目的:研究缺氧诱导因子1α(HIF-1α)在宫颈鳞状上皮癌变过程中的表达特点及意义。方法:采用免疫组化EliVision法检测HIF-1α在20例宫颈正常鳞状上皮、28例鳞状上皮不典型增生、12例宫颈原位癌和33例浸润性癌组织中的表达特点及其相关性。结果:在上皮恶性转化过程中,HIF-1α的表达逐渐增强,HIF-1α在浸润性癌与正常组织中的表达存在显著性差异(P<0.05);肿瘤伴淋巴结转移者HIF-1α表达明显高于未转移者(P<0.05)。结论:HIF-1α的过表达可能在宫颈鳞癌癌变及进展过程中发挥重要作用,并可作为宫颈鳞癌浸润转移的重要指标。

缺氧诱导因子-1α的表达与卵巢浆液性癌生物学行为的相关性

目的:探讨HIF-1α在卵巢浆液性癌发生和发展中的作用。方法:用免疫组织化学方法检测HIF-1α蛋白在52例人卵巢浆液性癌组织中的表达,分析HIF-1α与临床病理参数的关系。结果:HIF-1α蛋白在卵巢浆液性癌和卵巢交界性浆液性囊腺瘤中的高表达率明显高于正常卵巢组织和卵巢浆液性囊腺瘤,其差异有显著性(P<0.001);低分化、III期+IV期和腹水阳性HIF-1α蛋白的高表达率明显高于高中分化、I期+II期和腹水阴性者,差异具有显著性(P<0.05);HIF-1α蛋白表达与年龄无关,差异无显著性(P>0.05)。结论:HIF-1α与卵巢浆液性癌血管生成密切相关,在卵巢浆液性癌的发生、发展中起着重要作用。

Effect of acetyl L-carnitine on human retinal pigment epithelium-19 cells in hypoxic conditions

AIM:To investigate the effect of acetyl-L-carnitine(ALCAR)on cell viability,morphological integrity,and vascular endothelial growth factor(VEGF)expression in human retinal pigment epithelium(ARPE-19)cells using a hypoxic model.METHODS:In the first set of experiments,the optimal CoCl_(2) dose was determined by exposing ARPE-19 cell cultures to different concentrations.To evaluate the effect of ALCAR on cell viability,five groups of ARPE-19 cell culture were established that included a control group,a sham group(200μM CoCl_(2)),and groups that received 1,10 and 100 mM doses of ALCAR combined with 200μM CoCl_(2),respectively.The cell viability was measured by MTT assay.The morphological characteristics of cells were observed by an inverted phase contrast microscope.The levels of VEGF and HIF-1α secretion by ARPE-19 cells were detected by enzyme linked immunosorbent assay(ELISA)assay.RESULTS:ARPE-19 cells were exposed to different doses of CoCl_(2) in order to create a hypoxia model.Nevertheless,when exposed to a concentration of 200μM CoCl_(2),a notable decrease in viability to 83% was noted.ALCAR was found to increase the cell viability at 1 mM and 10 mM concentrations,while the highest concentration(100 mM)did not have an added effect.The cell viability was found to be significantly higher in the groups treated with a concentration of 1 mM and 10 mM ALCAR compared to the Sham group(P=0.041,P=0.019,respectively).The cell viability and morphology remained unaffected by the greatest dose of ALCAR(100 mM).The administration of 10 mM ALCAR demonstrated a statistically significant reduction in the levels of VEGF and HIF-1α compared with the Sham group(P=0.013,P=0.033,respectively).CONCLUSION:The findings from the current study indicate that ALCAR could represent a viable therapeutic option with the potential to open up novel treatment pathways for retinal diseases,particular relevance for age-related macular degeneration(AMD).However,to fully elucidate ALCAR’s application potential in retinal diseases,additional investigation is necessary to clearly define the exact mechanisms involved.

Hypoxia-inducible factor 1 alpha and vascular endothelial growth factor overexpression in ischemic colitis

AIM: To examine the etiology and pathophysiology in human ischemic colitis from the viewpoint of ischemic factors such as hypoxia-inducible factor 1 alpha (HIF-1 alpha and vascular endothelial growth factor (VEGF). METHODS: Thirteen patients with ischemic colitis and 21 normal controls underwent colonoscopy. The follow-up colonoscopy was performed in 8 patients at 7 to 10 d after the occurrence of ischemic colitis. Biopsy samples were subjected to real-time RT-PCR and immunohistochemistry to detect the expression of HIF-1 alpha and VEGF. RESULTS: HIF-1 alpha and VEGF expression were found in the normal colon tissues by RT-PCR and immunohistochemistry. HIF-1 alpha and VEGF were overexpressed in the lesions of ischemic colitis. Overexpressed HIF-1 alpha and VEGF RNA quickly decreased to the normal level in the scar regions at 7 to 10 d after the occurrence of ischemic colitis. CONCLUSION: Constant expression of HIF-1 alpha and VEGF in normal human colon tissue suggested that HIF-1 alpha and VEGF play an important role in maintaining tissue integrity. We confirmed the ischemic crisis in ischemic colitis at the molecular level, demonstrating overexpression of HIF-1 alpha and VEGF in ischemic lesions. These ischemic factors may play an important role in the pathophysiology of ischemic colitis.

Hypoxia inducible factor-1α mediates protective effects of ischemic preconditioning on ECV-304 endothelial cells

AIM: To investigate whether hypoxia inducible factor-1α (HIF-1α) is linked to the protective effects of ischemic preconditioning (IP) on sinusoidal endothelial cells against ischemia/reperfusion injury. METHODS: Sinusoidal endothelial cell lines ECV-304 were cultured and divided into four groups: control group, cells were cultured in complete DMEM medium; cold anoxia/warm reoxygenation (A/R) group, cells were preserved in a 4℃ UW solution in a mixture of 95% N2 and 5% CO2 for 24 h; anoxia-preconditioning (APC) group, cells were treated with 4 cycles of short anoxia and reoxygenation before prolonged anoxia- preconditioning treatment; and anoxia-preconditioning and hypoxia inducible factor-1α (HIF-1α) inhibitor (I-HIF-1) group, cells were pretreated with 5 μm of HIF-1α inhibitor NS398 in DMEM medium before subjected to the same treatment as group APC. After the anoxia treatment, each group was reoxygenated in a mixture of 95% air and 5% CO2 incubator for 6 h. Cytoprotections were evaluated by cell viabilities from Trypan blue, lactate dehydrogenase (LDH) release rates, and intracellular cell adhesion molecule-1 (ICAM-1) expressions. Expressions of HIF-1α mRNA and HIF-1α protein from each group were determined by the RT-PCR method and Western blotting, respectively. RESULTS: Ischemia preconditioning increased cell viability, and reduced LDH release and ICAM-1 expressions. Ischemia preconditioning also upregulated the HIF-1α mRNA level and HIF-1α protein expression. However, all of these changes were reversed by HIF-1α inhibitor NS398.CONCLUSION: Ischemia preconditioning effectively inhibited cold hypoxia/warm reoxygenation injury to endothelial cells, and the authors showed for the first time HIF-1α is causally linked to the protective effects of ischemic preconditioning on endothelial cells.

RNA interference against hypoxia inducible factor-1α inhibited growth of cervical cancer by limiting vascularization

Objective:Cervical cancer has become a major public health problem.The development of effective,systemic therapies for cervical cancer is highly desired.We show here that hypoxia inducible factor-1α(HIF-1α) was indicated as an attractive therapeutic molecular target for cervical cancer.Methods:Firstly,we observed the expressional level of HIF-1α in cervical cancer and Hela and Siha cell lines.Secondly,by constructuring HIF-1α shRNA targeting human HIF-1α mRNA common sequence and transfecting it with plasmid to cervical cell,we detected the changes of HIF-1α and its downstream genes levels VEGF.Then we injected selected stably transfected cell line into athymic nude mice to estimate its' antitumor effects.Results:We observed that HIF-1α inhibition was related to down-regulated VEGF resulting in prevention of angiogenesis,then leading to slower-growing tumors.Conclusion:The underlying concept of transfecting a HIF-1α shRNA expression vector to block the HIF-1α holds promise as the clinical potential of gene therapy for cervical cancer.

Electroacupuncture combined with intermittent pneumatic compression therapeutic apparatus for diabetic peripheral neuropathy and the effect on HIF-1α and VEGF levels

Objective： To observe the clinical efficacy of electroacupuncture combined with intermittent pneumatic compression therapeutic apparatus for treatment of diabetic peripheral neuropathy, and the effect on serum VEGF and HIF-lα levels of patients. Methods： Ninety-six patients were randomly divided into electroacupuncture treatment group （EA group）, intermittent pneumatic compression treatment group （IPC group）, electroacupuncture combined with intermittent pneumatic compression treatment group （EA ＋ IPC group） and cobamamide group （CM group）, with 24 cases in each group. Electroacupuncture treatment （once a day）, intermittent pneumatic com pression treatment （twice a day） and intramuscular injection with cobamamide （1 rag, once a day） were carried out in EA group, IPC group and CM group, respectively, and intermittent pneumatic compres- sion treatment （twice a day） was conducted on the basis of electroacupuncture treatment （once a day） in EA＋IPC group. After treatment for 2 consecutive weeks, the differences in subjective symptoms, mo- tor nerve conduction velocity, sensory nerve conduction velocity and serum HIF-lα and VEGF levels of patients in the four groups before and after treatment were observed and compared. Results： After treatment for 2 weeks, the differences in total effective rate between EA group and CM group, IPC group and CM group, as well as EA ＋ IPC group and CM group were all significant （all P 〈 0.05）, and the total effective rate in EA＋ IPC group was significantly higher than that in EA group and IPC group （both P 〈 0.05）. After treatment for 2 weeks, the motor nerve conduction velocity and sensory nerve conduction velocity of median nerve and common peroneal nerve of patients in EA group, 1PC group and EA＋IPC group were all higher than that before treatment （all P 〈 0.05）; the motor nerve conduction velocity of median nerve and the sensory nerve conduction velocity of common peroneal nerve in EA group were all higher than that in CM group （both P 〈 0.05）; the motor nerve conduction velocity and sensory nerve conduction velocity of median nerve in IPC group were also all higher than that in CM group （both P 〈 0.05）; the motor nerve conduction velocity and sensory nerve conduction velocity of median nerve and common peroneal nerve in EA＋IPC group were all higher than that in CM group （both P 〈 0.05）; the sensory nerve conduction velocity of common peroneal nerve in EA ＋ 1PC group was higher than that in EA group and IPC group （both P 〈 0.05）, and the motor nerve conduction velocity of median nerve in EA＋IPC group was higher than that in IPC group （P 〈 0.05）. The serum HIF-1α and VEGF levels of patients in EA group, IPC group and EA ＋ IPC group after treatment significantly reduced （all P 〈 0.05）. and were lower than that in CM group after treatment （all P 〈 0.05）; the serum HIF-lα and VEGF levels of patients in EA ＋ IPC group after treatment were lower than that in EA group and IPC group, and the difference in serum HIF-lα level was statistically significant （both P 〈 0.05）. Conclusion： Electroacupuncture combined with intermittent pneumatic compression therapeutic apparatus can effectively improve the clinical symptoms of patients with diabetic peripheral neuropathy, the efficacy were better than electroacupuncture, intermittent pneumatic compression treatment and cobamamide.

The effect of spironolactone on the levels of GATA4 protein and hypoxia inducible factor-1α in patients receiving coronary stent implantation

In the present study,we aimed to explore the protective effect of spironolactone on cardiac function in patients undergoing coronary stent implantation by determining the serum levels of GATA4 and hypoxia-inducible factor-1α(HIF-1α)proteins before and after the coronary stent implantation.A total of 134 patients undergoing coronary stent implantation in our hospital from March 2019 to March 2020 were retrospectively selected using the propensity score matching(PSM)method.Of the 134 patients,67 patients taking spironolactone were used as a test group,and the other 67 patients without taking spironolactone were used as a control group.In all patients,the levels of serum GATA4,HIF-1α,and troponin I proteins were determined before as well as 24 h and 6 months after the coronary stent implantation.Left ventricular ejection fraction(LVEF)and left ventricular systolic global longitudinal strain(GLS)were determined before and 6 months after the coronary stent implantation.There were no significant differences in the HIF-1αlevel between the two groups before and 6 months after the operation,while the HIF-1αlevel was significantly lower in the test group compared with the control group at 24 h after the operation(P<0.01).There were no significant differences in the GATA4 protein level between the two groups before and 24 h after the operation,while the GATA4 protein level was significantly lower in the test group compared with the control group at 6 months after the operation(P<0.01).There was no significant difference in LVEF between the two groups before and 6 months after the operation.GLS was significantly improved at 6 months after the operation compared with that before the operation in both groups,while GLS was significantly better in the test group compared with the control group at 6 months after the operation(P<0.01).Collectively,spironolactone could protect cardiac function probably by improving myocardial hypoxia and inhibiting myocardial remodeling.

Polycomb chromobox 4 enhances migration and pulmonary metastasis of hepatocellular carcinoma cell line MHCC97L

We recently report that the expression of polycomb chromobox 4(Cbx4)is significantly correlated with the overall survival of a great cohort of hepatocellular carcinoma(HCC)patients and it enhances hypoxia-induced vascular endothelial growth factor(VEGF)expression and angiogenesis in HCC cells through enhancing sumoylation of hypoxia inducible factor-1alpha(HIF-1α).Here we continue to investigate the potential effects of Cbx4 on the migration and metastasis of the metastatic HCC cell line MHCC97L.Our results show that Cbx4 overexpression in the cell line increases the in vitro vessel formation of vascular endothelial cells in its SUMO interaction motifs-dependent manner,and promotes the in vitro migration of the cancer cell,which can be effectively abrogated by anti-VEGF antibody.Although Cbx4 expression does not impact the in vitro growth of MHCC97L cells,it still promotes the progression and metastasis of orthotopically transplanted tumors in nude mice.These results further support the role of Cbx4 as a SUMO E3 ligase in the progression and metastasis of HCC.