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OBJECTIVE@#To investigate the effect of miR-204 on the invasion and metastasis of breast cancer by targeted regulation of HNRNPA2B1.@*METHODS@#The bioinformatics database was used to obtain data of the expressions of miR-204 in breast cancer patients and the survival rate of the patients. RT-qPCR was used to detect the expression of miR-204 in breast cancer cell lines. The expression vector GV369-miR-204 was used to overexpress miR-204 in MDA-MB-231 cells. Transwell assay was performed to detect the effect of miR-204 on the migration and invasion ability of the breast cancer cells. The key genes (hub genes) of miR-204 were determined by bioinformatics method. A dual luciferase assay was used to analyze the targeting relationship between miR-204 and HNRNPA2B1. The expression of HNRNPA2B1 in MDA-MB-231 cells after miR-204 overexpression was detected by Western blotting, and Transwell assay was used to examine the changes in the cell invasion ability.@*RESULTS@#The expression of miR-204 was decreased in both breast cancer tissues, and was significantly lower in breast cancer MDA-MB-231 cells than in MCF-10A cells ( < 0.05). The decreased expression of miR-204 was associated with poorer prognosis of breast cancer patients ( < 0.05). Upregulation of miR-204 in MDA-MB-231 cells significantly inhibited the invasion and migration of the cells ( < 0.05). Analysis of the data from the Starbase revealed that the expression of miR-204-5p was negatively correlated with the expression of HNRNPA2B1, and the expression of HNRNPA2B1 was increased in breast cancer patients ( < 0.05) in association with a poorer prognosis of the patients ( < 0.05). Dual luciferase assay demonstrated that miR-204 could bind to HNRNPA2B1 in a target-specific manner. Western blotting and Transwell assay showed that miR-204 significant inhibited the migration and invasion ability of breast cancer cells by targeting HNRNPA2B1 ( < 0.05).@*CONCLUSIONS@#miR-204 expression is decreased in breast cancer tissues and cells, and its overexpression can inhibit the invasion and metastasis of breast cancer cells by targeted regulation of HNRNPA2B1.
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Humans , Breast Neoplasms , Cell Line, Tumor , Cell Movement , Cell Proliferation , Gene Expression Regulation, Neoplastic , MicroRNAs , Genetics , Neoplasm Invasiveness , Neoplasm MetastasisABSTRACT
BACKGROUND@#Derlin 3 (DERL3) is downregulated in colorectal cancer (CRC) samples. Its level is closely linked to lymphatic metastasis or distant metastasis rate in CRC patients. However, its biological behavior in lung adenocarcinoma were rarely reported. The aim of this study is to investigate the ectopic expression of DERL3 in lung adenocarcinoma tissues and its effect on the invasion and metastasis of lung adenocarcinoma A549 cell line to reveal the possible mechanism of invasion and metastasis of lung adenocarcinoma.@*METHODS@#Lung adenocarcinoma microarray gene chip data included 3 cases of lymph node metastasis and 3 cases of lung adenocarcinoma tissue without lymph node metastasis. The GEDS and Kaplan-Meier plot queries the survival curve and expression level of DERL3. Western blot was used to detect the expression of DERL3 in lung adenocarcinoma cells. The efficiency of knockdown DERL3 gene was detected by Western blot assay. Transwell detected the number of cells passing through the basement membrane of the transwell. EDU assay detected cell proliferation ability. Western blot detected the expression of epithelial-mesenchymal transition related proteins E-cadherin and Vimentin.@*RESULTS@#The microarray gene chip results showed that compared with lung adenocarcinoma tissues without lymph node metastasis, 1,314 mRNAs in lung adenocarcinoma tissues with lymph node metastasis were up-regulated, 400 mRNAs were down (P<0.05). The expression of DERL3 increased in lung adenocarcinoma (P<0.05). The results of survival curve showed that the lung cancer patients with high expression of DERL3 with poor prognosis (P<0.05). Western blot results indicated that plasmid transfection was successful. Knockdown of DERL3 suppressed the ability of proliferation, invasion and migration in A549 cells (P<0.05). After knockdown of DERL3, the expression level of Vimentin was decreased, while E-cadherin expression increased (P<0.05).@*CONCLUSIONS@#Knockdown of DERL3 inhibited the proliferation, invasion and metastasis of A549 cells.
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ObjectiveTo investigate the risk factors for acute renal injury (AKI) in patients with ischemic stroke.MethodsPatients with ischemic stroke were enrolled retrospectively.The general clinical data, vascular risk factors, drug use, stroke etiological typing, stroke severity, and baseline biochemical indices were collected.They were divided into either an AKI group or a control group according to whether AKI occurred or not.Multivariable logistic regression analysis was used to analyze the independent risk factors for occurring AKI in patients with ischemic stroke.ResultsA total of 214 patients with ischemic stroke were enrolled, including 32 (14.95%) had AKI and 182 (85.05%) did not have AKI.The proportions of patients in heart failure (62.50% vs.41.21%;χ2=4.998, P=0.025), mannitol use (87.50% vs.43.96%;χ2=20.643, P200 ml (28.13% vs.9.89%;χ2=6.637, P=0.010), as well as NIHSS score (18.0±4.5)vs.8.0±3.2;t=15.249, P200 ml (OR 3.294, 95% CI 1.464-2.786;P=0.021) were the independent risk factors for AKI in patients with acute ischemic stroke.ConclusionsThe NIHSS score, diastolic blood pressure, arterial lactate concentration,mannitol use, furosemide use, contrast agent use and contrast dosage >200 ml were associated with AKI in patients with ischemic stroke.
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Objective To investigate the effects of N-acetylserotonin (N-AS) on the expression of active caspase-3,Bcl-2 and Bax in rat retinas induced by retinal ischemia-reperfusion injury (RIRI).Methods Adult male Sprague-Dawley rats were randomly divided into the normal control group (6 cases),RIRI group (30 cases) and NAS group (30 cases),RIRI models in NAS group were established after giving NAS,the groups were sub-divided into 6 hours,12 hours,24 hours,48 hours and 72 hours group based on the time of RIRI.Morphologic changes were evaluated by HE staining.The expression of active caspase-3,Bcl-2 and Bax protein in the retina of rats was detected by immunohistochemistry.Results HE staining showed that the retinal structure in the normal control group was clear,and the cells in each layer were tightly packed;Each layer of retina was edema in the RIRI group after 6 hours and 12 hours,the edema gradually alleviated after 24 hours,the ganglion cells decreased gradually,the distribution was in disorder,with the prolongation of time,the retinal ganglion cells were defected;drug group of as Compared with RIRI group,the cell edema in the NAS group at 6 hours and 12 hours were obvious reduced,the cells in 24 hours,48 hours,72 hours group arranged regularly,the loss number of ganglion cells were reduced.The number of active caspase-3 positive cells in RIRI group increased at 6 hours after peffusion,the number was (561.15 ±37.19) cell ·mm-2,and reached the high level at 24 hours,the number was (1522.61 ±84.36) cell · mm-2,and then decreased gradually.The number of active caspase-3 positive cells in NAS group was significantly lower than that in RIRI group,the difference was statistically significant (all P ＜ 0.05).The expression of Bcl-2 positive cells in RIRI group began to decrease after 6 hours,and decreased to a low level at 24 hours,and the number of Bcl-2 positive cells in NAS group was significantly higher than that in RIRI group at each time point,the differences were statistically significant (all P ＜ 0.05).There were almost no Bax positive cells in the retina of the control normal group,and the Bax positive cells were found to be higher of the RIRI group at the 6 hours after RIRI,and reached the higher level at 24 hours,and decreased at 48 hours.The Bax positive cells of NAS group were significantly less than those in the RIRI group at different time points,and the differences were statistically significant (all P ＜0.05).Conclusion NAS can promote the expression of Bcl-2 protein in rat retina after RIRI,inhibit the expression of Bax protein,decrease the expression of active caspase-3 protein,alleviate cell apoptosis,and have neuroprotective effects.
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Objective Based on middle cerebral artery occlusion (MCAO) model,to investigate the effects of melatonin (MT) on DWI and expression of Fas,FasL and cleaved Caspase-3 proteins in rat model with focal cerebral ischemia.Methods Eighty SD rats were randomly divided into Sham group (n=16),MCAO group (n=32) and MT group (n=32).The rats in sham group were treated with sham-operation.And the rats in MCAO and MT groups were peritoneally injected with saline and MT respectively.The behavioral scores were assessed in the three groups.The rats in MCAO and MT group with the behavioral scores of 1 3 points were selected in the study.The DWI relative signal intensity (rDWI-SI),Fas,FasL and cleaved Caspase-3 proteins were respectively examined by MR scaning and immunohistochemical staining in all rats of each group at 6 h,24 h,72 h and 7 days after ischemia reperfusion (IR) or sham-operation.And the DWI and immunohistochemical results for each group were compared.Results At last,there were 16 rats in sham group,29 rats in MCAO group and 30 rats in MT group,respectively.There was significant difference of the behavioral scores among the three groups (x2 =50.125,P＜0.01).The behavioral scores of MT and MCAO groups were higher than those of sham group (all P ＜0.05).And the behavior scores of the MT group were lower compared with MCAO group after IR.Compared with the rDWI-SI values measured at 6 h,24 h and 72 h,7 days in sham group,the rDWLSI values of MT and MCAO groups were significantly higher (all P＜0.01).And the rDWI-SI was higher in MCAO group than those in MT group at 6 h,24 h and 72 h after IR (all P＜0.01).And there was no significant difference of rDWI-SI at 7 days after IR between MT and MCAO groups (P＞0.05).The immunohistochemical staining results showed that the number of Fas,FasL and cleaved Caspase-3 positive cells in MCAO and MT groups were significantly higher than those in sham group (all P＜0.01).And there were less Fas,FasL and cleaved Caspase-3 positive cells in MT groups compared with MCAO group (all P＜0.05) at 6 h,24 h and 72 h after IR.There was no significant difference of Fas,FasL and cleaved Caspase-3 positive cells among the three groups at 7 days after IR (P＞0.05).Conclusion MT can effectively alleviate the rDWI-SI value and inhibit the expression of Fas,FasL and cleaved Caspase-3 proteins in rats of focal cerebral ischemia.
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BACKGROUND:cAMP response element binding protein (CREB) is a key protein of memory, which is closely related to long-term memory. It wil provide a new way for the treatment of hypoxic ischemic brain damage (HIBD) to study the effects of dental pulp stem cel s transplantation on the long-term behavior and CREB protein via the lateral ventricle in neonatal HIBD rats. OBJECTIVE:To observe the changes in long-term behavior and CREB protein expression in neonatal HIBD rats after human dental pulp stem cel transplantation, thereby providing scientific evidence for clinical treatment of neonatal HIBD. METHODS:Thirty-six healthy 7-day-old Sprague-Dawley rats were randomly divided into normal, HIBD and cel transplantation group. The hypoxic ischemic brain damage models were established in the brain damage and cel transplantation groups. Twenty-four hours after HIBD, human dental pulp stem cel s were injected into the left lateral cerebral ventricle of rats in the cel transplantation group, total y 3×106 living cel s. Equal volume of normal saline was injected into the left lateral cerebral ventricle of rats in the normal control and HIBD groups. RESULTS AND CONCLUSION:The average time to seek water, the average escape latency and escape distance of the human dental pulp stem cel s group were significantly shorter than those of hypoxic ischemic brain injury group (P<0.01), but longer than those in the normal group (P<0.01). Nissl staining showed that the cel s in the hippocampal CA1 region in human dental pulp stem cel s group were more regular, the number of cel s was significantly higher than that of hypoxic ischemic brain injury group, but stil significantly less than that in the normal group (P<0.05). Immunohistochemical staining results showed that the number of CREB positive cel s in human dental pulp stem cel s group was significantly higher than those in HIBD group, but stil significantly less than those in the normal group (P<0.01). It is suggested that human dental pulp stem cel s transplantation could promote the expression of CREB protein in the hippocampal CA1 region, to improve the long-term learning and memory ability of hypoxic ischemic neonatal rats, and thus repair HIBD.