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@#[摘 要] 目的：探讨α-常春藤皂苷（α-Hed）诱导非小细胞肺癌（NSCLC）细胞凋亡的作用靶点及其潜在机制，明确α-Hed与顺铂（DDP）联用后对相应的靶点蛋白表达的影响。方法：采用CCK-8法检测不同浓度α-Hed处理后NSCLC细胞A549、H1299和PC-9的存活率，采用Annexin Ⅴ-FITC/PI染色流式细胞术检测细胞凋亡率，采用WB法检测细胞中C-caspase-3和Bcl-2蛋白的表达。通过网络药理学相关方法筛选α-Hed的潜在靶点，利用分子对接法分析其结合效果，WB法检测靶点蛋白的表达。通过CCK-8法、细胞集落形成实验和WB法检测α-Hed与DDP联用对NSCLC细胞的抑制作用。结果：给药24和48 h后，10、15和20 μmol/L α-Hed可以显著抑制NSCLC细胞增殖活力（均P<0.01）；与对照组相比，20 μmol/L α-Hed处理后细胞凋亡率显著升高（P<0.01）；α-Hed可上调NSCLC细胞中C-caspase-3的表达（P<0.05），下调Bcl-2的表达（P<0.05）。网络药理学和分子对接筛选出结合亲和力小于-5 kcal/mol的靶点AKT1、STAT3、EGFR和JAK2。WB法检测结果显示，α-Hed处理后A549、H1299细胞中EGFR、p-AKT/AKT、p-STAT3/STAT3和JAK2蛋白的表达均明显下调（均P<0.05）。α-Hed与DDP联用后，更显著地抑制NSCLC细胞的增殖（P<0.01），进一步下调EGFR、p-AKT/AKT、p-STAT3/STAT3和JAK2蛋白的表达（P<0.05或P<0.01）。结论：α-Hed通过下调EGFR和JAK2的表达抑制STAT3和AKT的磷酸化，诱导NSCLC细胞凋亡，与DDP联用后其抑制效果增强，EGFR/AKT和JAK2/STAT3通路也进一步被抑制。
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AIM: To explore the mechanism of fructus lycii in treating dry eye based on network pharmacology and experimental verification.METHODS: Taking “fructus lycii” as key words, the active ingredients and target of fructus lycii were searched by using Traditional Chinese Medicine Systems Pharmacology Database and Analysis Platform(TCMSP). Gene targets related to dry eye(DE)were searched by GeneCards and OMIM databases. The target genes of fructus lycii and DE were imported into Venn software to obtain the intersection target map of them. After that, the data were imported into the String database to obtain the PPI protein-protein interaction network diagram. Using Cytoscape3.7.2 software, the PPI protein-protein interaction network diagram was constructed for active ingredients, target sites and related diseases of fructus lycii. The Bioconductor platform and R language were used for gene ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment analysis. And the key targets in the pathogenesis of DE were verified by experiments.RESULTS: Through TCMSP, 45 types of effective chemical components of fructus lycii, 174 target genes corresponding to active components and 131 common target genes with DE were screenedout. In accordance with the network topology of “drug-composition-disease-target”, 27 main effective components of fructus lycii were found in the treatment of DE. The PPI network was analyzed according to the high degree value, which is the key targets of fructus lycii for DE treatment, mainly including AKT1, VEGFA, CASP3, IL1B, JUN, PTGS2, CXCL8, etc. According to GO enrichment analysis, 166 biological functions and processes of fructus lycii for DE treatment were obtained. KEGG enrichment analysis showed that 31 signaling pathways were involved. Additionally, experimental verification displayed that the protein expressions of AKT1, interleukin-6(IL-6), tumor necrosis factor(TNF-α)and IL-17 in conjunctiva tissue of the DE model group were significantly increased.CONCLUSIONS: Through network pharmacology, this study confirmed that the treatment of DE by fructus lycii is a complex process involving multi-components, multi-targets and multi-pathways, and that the treatment of DE by fructus lycii is mainly regulated by anti-inflammatory and apoptosis-related molecules.
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Proteus syndrome is a rare congenital hamartomatous syndrome characterized by the asymmetric and disproportionate overgrowth of limbs, emergence of connective tissue nevi, epidermal nevi, ysregulated adipose tissue, and vascular malformations. The Proteus syndrome is caused by mosaicism of somatic activating mutation in the AKT1 gene which locates at chromosome 14q32.3. This syndrome is extremely rare, making it difficult to diagnose. The most commonly used diagnostic criteria are too complicated to be used in clinical practice. Surgery can partially alleviate the clinical symptoms of overgrowth, but it can't inhibit the progression of the disease. This article summarizes the diagnostic criteria, treatment principles, and perioperative managements for Proteus syndrome in the world. The article proposes the highly suspected morphological manifestations of Proteus syndrome was based on clinical experiences of the author.The article emphasizes using genetic detection of pathological tissue as the gold standard for diagnosis, and suggests targeted therapy as the optimal treatment for Proteus syndrome.
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AIM: To explore the potential mechanism of action of Pingxiao capsule in the treatment of breast cancer. METHODS: TCMSP, TCM-ID, GeneCards and other databases were used to screen the related targets of Pingxiao capsule and breast can-
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Objective:To explore the value of single nucleotide polymorphism for molecular warning of hypertensive intracerebral hemorrhage in Li Nationality in Hainan.Methods:Totally 56 patients with hypertensive intracerebral hemorrhage of Li Nationality in Hainan Province, 100 healthy controls of Li nationality and 203 healthy controls of Han nationality in Hainan Province from January 2019 to October 2020 were selected as the research subjects.After genomic DNA was extracted, rs2494739, rs2494744 and rs2498794 of AKT1 gene were genotyped and analyzed by SPSS 25.0 to explore the differences between Han and Li Nationality, and between Li healthy population and intracerebral hemorrhage population.Results:There was no difference in the frequencies of rs2498794, rs2494739 and rs2494744 polymorphisms of AKT1 gene among Han and Li healthy controls ( P>0.05). The rates of AA, AG and GG at rs2498794 locus in Li Nationality patients with intracerebral hemorrhage (14.28%, 39.29% and 46.43%) were significantly different from those of Li control group (44.00%, 47.00% and 9.00%)( P<0.05). The distribution rates of AA, AG and GG of rs2494744 in Li Nationlity patients with intracerebral hemorrhage were 57.14%, 37.50% and 5.36%, respectively, which were statistically significant compared with the control group (20.00%, 44.00% and 36.00%) ( P<0.05). The incidence of CC, CT and TT at rs2494739 locus in Hainan Li Nationality patients were 14.28%, 46.43% and 39.29% respectively, which were also significantly different from those in Li control group(34.00%, 41.00% and 25.00%) ( P<0.05). The incidence of rs2494744-A in intracerebral hemorrhage group (75.89%) was much higher than that in Li control group (42.00%), and the OR value of rs2494744-A was 4.35.The incidence of rs2498794-G in intracerebral hemorrhage group and control group were 66.07% and 32.50%, respectively, and the OR was 4.04.Alleles rs2494744-A and rs2498794-G were moderately associated with the incidence of intracerebral hemorrhage ( P<0.05). Conclusion:rs2494744-AA, rs2498794-GG and alleles rs2494744-A and rs2498794-G are the risk factors of HICH in Li nationality, which is of great value to the construction of its molecular early warning system.
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Objective @#To investigate the expression and prognosis of urokinase plasminogen activator (PLAU) and v-akt murine thymoma viral oncogene homolog 1 (AKT1) in oral squamous cell carcinoma ( OSCC) and normal tis- sues and the correlation of PLAU and AKT1 in OSCC tissues．@*Methods @#The expression levels of PLAU and AKT1 in 70 cases of oral squamous cell carcinoma and 50 cases of normal tissues were detected by immunohistochemical method，and the correlation between PLAU and AKT1 expression and clinicopathological features and prognosis as well as the correlation between PLAU and AKT1 expression in OSCC tissues was analyzed，and the results were fur- ther verified by bioinformatics database. @*Results @#The expression of PLAU and AKT1 in OSCC tissues was higher than that in normal tissues (P＜0. 05) ，Kaplan-Meier analysis showed that patients with low PLAU and AKT1 ex- pression had longer survival time than those with high AKT1 expression (P＜0. 05) ，Spearman rank sum correlation test showed that there was a strong correlation between PLAU and AKT1 expression in OSCC tissues (r = 0. 357，P ＜0. 05) ，GEPIA bioinformatics database analysis results are consistent with experimental results．@*Conclusion@#PLAU and AKT1 are highly expressed in OSCC tissues and are associated with poor prognosis of patients．There is a correlation between PLAU and AKT1 in OSCC tissues.
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Objective:To investigate the neuroprotective effects of Danggui Shaoyaosan （DSS） on APP<sub>swe</sub>/PS1<sub>ΔE9 </sub>transgenic （APP/PS1） mice and its mechanism related to circular RNA （circRNA）. Method:Totally twenty 6-month-old APP/PS1 mice were divided into model group and DSS group， and 10 C57BL/6 wild-type mice were set as the normal control group. The normal group and model group received the same volume of normal saline， and DSS group received drug by gavage administration， all for 8 weeks. The differentially expressed circRNA of APP/PS1 mice before and after DSS intervention was analyzed by circRNA sequencing to construct circRNA-miRNA mRNA interaction network. The results of cricRNA sequencing were then verified by Real-time fluorescence quantitative polymerase chain reaction （Real-time PCR）. The protein expression levels of phosphoinositide 3-kinase （PI3K）， p-PI3K， protein kinase B1 （Akt1）， p-Akt1， B lymphocytoma-2 （Bcl-2）， and Bcl-2-Associated X protein （Bax） in the hippocampus were detected by immunoblotting （Western blot）. The protein expression of Caspase-3 in the hippocampus was detected by immunohistochemistry and the level of apoptosis in the hippocampus was detected by the TUNEL method. Result:Compared with the model group， there were 90 differentially expressed circRNA after intervention with DSS， of which 46 were up-regulated and 44 down-regulated. Compared with the normal group， the expression levels of circRNA1398 and circRNA1399 in the model group decreased， and the expression levels of miR-103-3p， miR-153-3p， miR-143-3p， and miR-143-5p increased. Compared with the model group， the expression levels of circRNA1398 and circRNA1399 in the DSS group were up-regulated， while the expression levels of miR-103-3p， miR-153-3p， miR-143-3p， and miR-143-5p were down-regulated. Compared with the normal group， the expression of p-PI3K， Akt1， p-Akt1， and Bcl-2 in the model group decreased （<italic>P</italic><0.05，<italic> P</italic><0.01）， and the expression of Bax and Caspase in the model group increased （<italic>P</italic><0.01）. Compared with the model group， the expression of p-PI3K， Akt1， p-Akt1， and Bcl-2 in the hippocampus of the DSS group increased （<italic>P</italic><0.01）， and the protein expression of Bax and Caspase decreased （<italic>P</italic><0.01）. Compared with the normal group， the apoptosis level in the hippocampus of the model group increased， with an apoptosis rate of （43.76±2.92）%. Compared with the model group， the apoptosis rate of DSS group was （24.64±3.39）%. Conclusion:DSS can activate PI3K/Akt pathway and inhibit apoptosis in hippocampal neurons of APP / PS1 mice， and play a neuroprotective role. The specific mechanism may be related to the regulation of circRNA1398 and circRNA1399 expression and the corresponding miRNA expression.
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Objective:To investigate the early warning effect of two polymorphisms of AKT1 gene (rs1130214, rs2494744) on the risk of atherosclerotic cerebral infarction in Hainan province.Methods:In this study, totally 243 patients with atherosclerotic cerebral infarction who were hospitalized of Hainan Province from January 2019 to October 2020 were selected as the cerebral infarction group, including 148 Han nationality people and 95 Li nationality people. And 272 healthy people who received physical examination in the same hospital during the same period were selected as the control group, including 197 Han nationality people and 75 Li nationality people.All participants signed informed consent. The peripheral anticoagulant DNA was collected, and the genomic DNA was extracted and amplified by PCR.The genotypes of rs1130214 and rs2494744 were analyzed by mass spectrometry, and the distribution frequency of genotypes in cerebral infarction group and control group was analyzed by Chi-square test with SPSS 25.0 software.Results:Regardless of nationality, there was no significant difference in the distribution frequency of rs1130214 and rs2494744 of AKT1 gene between cerebral infarction group and control group (both P>0.05). The frequencies of AA, AG and GG genotypes of rs2494744 locus were 44.59%, 51.36% and 4.05% in the cerebral infarction group of Han nationality, while they were 47.21%, 42.13% and 10.66% in the control group of Han nationality, with significant difference between the two groups(χ 2=6.396, P<0.05). The independent effects of the three genotypes were analyzed by regression analysis. The results showed that GG genotype might be a resistance factor of cerebral infarction in Han population ( P=0.024, OR=0.354, 95% CI: 0.139-0.901). The frequency of AA, AG and GG genotypes of rs2494744 was 58 (61.05%), 25 (26.32%), 12 (12.63%) in the control group of Li nationality, and 28 (37.33%), 39 (52.00%), 8 (10.67%) in the control group of Li nationality. The results showed that the distribution of AA was significantly higher than that of the control group ( P<0.05, OR= 2.631, 95% CI=1.410-4.09), while AG was on the contrary ( P<0.05, OR=0.330, 95% CI=0.173-0.627). Conclusion:AA genotype of rs2494744 in AKT1 gene polymorphism is a risk factor for cerebral infarction in Li nationality group, which has potential early warning value for cerebral infarction in Hainan Li nationality group, while AG has protective effect on cerebrovascular health in Hainan Li nationality group.
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Objective:To investigate the effect of GluA2-3Y which is an inhibitor of AMPA(α-amino-3-hydroxy-5-methyl-4-isoxazole-propionic acid) receptor internalization on cognitive function and hippocampal postsynaptic protein expression in rats with chronic cerebral hypoperfusion.Methods:Forty-eight adult male SD rats were randomly divided into Sham group, 2VO group, high-dose GluA2-3Y group and low-dose GluA2-3Y group according to random number table, with 12 rats in each group.The chronic cerebral hypoperfusion model of rat was established by two vessel occlusion (2VO) while the Sham operation was performed in rats of Sham group.The rats in high dose GluA2-3Y group and low dose GluA2-3Y group were intraperitoneal injected with 3 μmol/kg and 0.03 μmol/kg GluA2-3Y respectively once a day for 2 weeks. Rats in 2VO group and Sham group were intraperitoneally injected with control peptide. Morris water maze test and new object recognition test were performed to evaluate the learning and memory ability of rats, and Western blot was used to evaluate the expression of Akt1、GSK3β、p-GSK3β、GluA2 and PSD-95 in rat hippocampus. The expressions of GluA2 and PSD-95 in rat hippocampus were evaluated by immunofluorescence. SPSS 23.0 software was used for data analysis. The comparison between multiple groups was analyzed by one-way ANOVA and repeated measurement ANOVA was used to analyze Morris water maze results. And independent-samples t-test was used for pairwise comparisons. Results:(1)In Morris water maze trials, the results of repeated measurement ANOVA showed that the interaction between group and time of escape latency of rats in each group was not significant ( F=0.79, P>0.05), and the group main effect and time main effect were significant ( F=24.44, 40.42, both P<0.05). On the 5th day of navigation trials, the escape latency of rats in 2VO group was longer than that in sham group ( t=5.87, P<0.05). The escape latency of rats in low dose GluA2-3Y group and high dose GluA2-3Y group were significantly shorter than that in 2VO group ( t=2.20, 3.41, both P<0.05), but there was no significant difference between low dose GluA2-3Y group and high dose GluA2-3Y group ( t=1.37, P>0.05). The target quadrant residence time and resolution coefficient ((14.57±1.40)s, (0.15±0.10)) in 2VO group were significantly lower than those in Sham group ((23.71±2.57)s, (0.40±0.06)) ( t=3.23, 2.24, both P<0.05), while the target quadrant residence time in high dose GluA2-3Y group ((20.19±1.53)s) and low dose GluA2-3Y group ((20.31±2.06)s) were longer than that in 2VO group( t=2.71, 2.35, both P<0.05). The discrimination coefficients in high dose GluA2-3Y group (0.47±0.10) and low dose GluA2-3Y group (0.59±0.06) were higher than that of 2VO group ( t=2.21, 3.94, both P<0.05). (2)The Western blot results showed that the expression of PSD-95 and GluA2 in hippocampus of rats in 2VO group were significantly lower than those in Sham group ( t=2.31, 2.20, both P<0.05), and the expression of PSD-95 in high dose GluA2-3Y group (1.026±0.056) was significantly higher than that in 2VO group ((0.760±0.061), t=2.49, P<0.05), while there was no significant difference between low-dose GluA2-3Y group and 2VO group( t=0.96, P>0.05). The expression of GluA2 in low-dose GluA2-3Y group was higher than that in 2VO Group ((1.130±0.087), (0.766±0.080), t=2.37, P<0.05), but there was no significant difference between high-dose GluA2-3Y group and 2VO group( t=1.06, P>0.05). (3) Immunofluorescence showed that compared with Sham group, the expression of PSD-95 and GluA2 in 2VO group decreased ( t=4.23, 2.57, P<0.05). Compared with 2VO group, the expression of PSD-95 and GluA2 in high dose GluA2-3Y group and low dose GluA2-3Y group increased significantly, and the differences were statistically significant (PSD-95: (7.757±0.578), (12.057±0.578), t=3.14, 6.96, both P<0.05; (9.721±0.950), (16.610±0.950), t=4.56, 9.34, both P<0.05). (4) The results of Western blot showed that the expression GSK3β in hippocampus of rats in each group were not statistically different( F=2.03, P>0.05). There were significant differences in the expression of Akt1, p-GSK3β and the percentage of p-GSK3β/GSK3β in hippocampus of rats in each group ( F=8.30, 4.76, 3.57, all P<0.05). Compared with Sham group, the levels of Akt1, p-GSK3β and the percentage of p-GSK3β/GSK3β in 2VO group were significantly lower ( t=3.00, 2.81, 3.17, all P<0.05). Compared with 2VO group, the levels of Akt1, p-GSK3β and p-GSK3β/GSK3β percentage in low dose GluA2-3Y group and high-dose GluA2-3Y group were significantly higher (Akt1: t=2.05, 5.20, both P<0.05; p-GSK3β: t=2.49, 4.15, both P<0.05; p-GSK3β/GSK3β percentage: t=2.30, 2.97, both P<0.05). Conclusion:GluA2-3Y, an AMPA receptor internalization inhibitor, can alleviate the cognitive impairment in rats with chronic cerebral hypoperfusion, which may be related to the increased expression of Akt1, p-GSK3β and postsynaptic proteins.
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Objective: To probe active components of Scutellariae Barbatae Herba (SBH) and its underlying complex mechanism in treating pancreatic cancer based on network pharmacology and molecular experimental validation. Methods: Active compounds of SBH and potential targets of these compounds were screened via Traditional Chinese Medicine Systems Pharmacology (TCMSP) database. Then the pancreatic cancer target database was established by using Comparative Toxicogenomics Database (CTD), Therapeutic Target Database (TTD), and Pharmacogenomics Knowledgebase (PharmGKB). Based on the matching results between SBH potential targets and pancreatic cancer targets, a PPI network was structured and then the hub targets were screened by using topology analysis. Furthermore, DAVID bioinformatics was utilized for both Gene ontoloty (GO) functional enrichment analysis and KEGG pathway enrichment analysis. At last, Western blotting was used to validate the regulating function of luteolin on P53, Bax and Bcl-2 targets of PANC-1 cells. Results: A total of 28 active ingredients including quercetin, luteolin and wogonin were screened out, and 24 hub targets of 91 targets including TP53, AKT1, JUN and VEGF were obtained. The results of DAVID enrichment analysis indicated that 73 cellular biological processes and 18 pathways were found to participate in the treatment of pancreatic cancer, mainly involving in regulating inflammatory microenvironment, cell cycle arrest, pro-apoptosis and anti-angiogenesis through NF-κB, p53, PI3K-AKT and VEGF signal pathways. Luteolin from SBH was validated to regulate key targets in the p53 signaling pathway to play a role in pro-apoptosis, which proved the reliability of results of network pharmacology. Conclusion: This study confirmed the synergistic effect of multiple component-multiple target-multiple pathway of SBH on the treatment of pancreatic cancer, which offered a theory for its clinical application.
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Objective To investigate the expression of miR-34a-5p and AKT1 genes in endometrium tissues of patients with endometriosis (EM) and their effects on migration and invasion of endometrial stromal cells (ESCs). Methods A total of 91 patients, undergone hysterectomy in the Department of Obstetrics and Gynecology of Zhujiang Hospital of Southern Medical University from Jan. 2018 to Jun. 2019 due to benign gynecological diseases, were collected and divided into EM group (68 cases) and non-EM group (23 cases). The expressions of miR-34a-5p and AKT1 genes in endometrium tissues of patients were detected by in situ hybridization and immunohistochemistry. ESCs were transfected with miR-34a-5p mimic and negative control RNA (miR-NC) using liposome-3000 to construct the cell models of miR-34a-5p mimic group and miR-NC group. The cell proliferation rate was detected by CCK-8 method, and cell migration, invasion, apoptosis and autophagy ability experiments were performed to determine the effect of miR-34a-5p on ESCs' proliferation, migration, invasion, apoptosis and autophagy. Results The positive expression rate of miR-34a-5p was lower, and of AKT1 was higher in EM group than those in non-EM group (16.2% vs. 82.6%, X2=34.323; 72.1% vs. 30.4%, X2=12.581, P<0.001). After culturing for 12, 24 and 48 h, the cell proliferation rate was higher in miR-NC group than that in miR-34a-5p mimic group (P<0.05). The cell migration ability and invasion ability were higher in miR-NC group than those in miR-34a-5p mimic group with statistically significant difference [(65.00%±5.00%) vs. (30.67%±4.04%); (88.0±8.5) vs. (32.3±6.1), t=9.179, P<0.05]. The cell apoptosis rate and the expression level of LC3 gene were obviously lower in miR-NC group than those in miR-34a-5p mimic group [(9.33%±3.51%) vs. (18.00%±2.00%); (0.19±0.04) vs. (0.39±0.03), t=8.02, P<0.05]. Conclusion miR-34a-5p may be involved in the pathogenesis of EM by targeting AKT1 genes to affect the proliferation, migration, invasion, apoptosis and autophagy function of ESC.
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PURPOSE: The prevalence of PIK3CA in Chinese breast cancer patients may be underestimated. Therefore, we investigated the distribution of somatic PIK3CA/AKT1 mutations in Chinese breast cancer patients and explored their roles in tumor phenotypes and disease prognosis. MATERIALS AND METHODS: Tumors from 507 breast cancer patients were prospectively collected from the West China Hospital between 2008 and 2013. Whole exons of AKT1 and PIK3CA were detected in fresh-frozen tumors using next-generation sequencing, and correlations between PIK3CA/AKT1 mutations and clinicopathological features were analyzed. RESULTS: The AKT1 mutation was found in 3.6% (18/507) of patients. Tumors from patients that carried the AKT1 mutation were estrogen receptor (ER)+/progesterone receptor (PR)+/human epidermal growth factor receptor 2 (HER2)‒ and were more likely to have high expression levels of Ki67. The prevalence of the PIK3CA mutation was 46.5% (236/507), and 35 patients carried two or three variants of the PIK3CA gene. PIK3CA mutations were associated with ER+/PR+/HER2‒ status. The prognosis of patients with one mutation in PIK3CA (or PIK3CA/AKT1) was not significantly different than that of patients with wild-type PIK3CA (or PIK3CA/AKT1), while patients with two or three variants in PIK3CA (or PIK3CA/AKT1) exhibited poorer outcomes in the entire group and in all three subgroups (ER+, HER2‒, Ki67 high), particularly with respect to overall survival. CONCLUSION: A high frequency of somatic PIK3CA mutations was detected in Chinese breast cancer patients. In addition to the mutation frequency, the tumor mutational burden of the PIK3CA and AKT1 genes should also be of concern, as they may be associated with poor prognosis.
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Humans , Asian People , Breast Neoplasms , Breast , China , Estrogens , Exons , Mutation Rate , Phenotype , Prevalence , Prognosis , Prospective Studies , ErbB ReceptorsABSTRACT
Objective Proteus syndrome is a rare disease. The aim of the present study was to analyze the clinical characteristics and gene mutations of Proteus syndrome with a case report and relevant literature review. Methods Clinical data of the patient with Proteus syndrome were collected in detail and biochemical measurements and radiological examinations were conducted. Tissues from phalanges with lesions were obtained to extract DNA, and Sanger sequencing of AKT1 gene was carried on. The pathogenic mutation was further tested in peripheral blood samples of the patient, his parents and 250 healthy volunteers. Orthopaedic surgery was performed on the affected limbs of the patient. Results The patient was presented with progressive overgrowth of the right extremity, scoliosis, cerebral connective tissue nevus and lower extremity venous. A heterozygous mutation of AKT1 gene (c. 49G>A) was identified in DNA extracted from the affected bone tissue of the patient, but not be found in genomic DNA of peripheral blood samples from the patient, his parents and 250 healthy volunteers. Movement function of the affected limb improved significantly after the operations. Conclusions The prominent features of Proteus syndrome are overgrowth of one extremity and cerebral connective tissue nevus. A mosaic somatic mutation of AKT1 gene is one of the pathogenic mutations for Proteus syndrome, and orthopedic surgery may be a good way to improve symptoms of the disease.
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Objective To investigate the effect of sodium houttuyfonate on the expression of PI3K and AKT1 and mTOR mRNA in the lung of rats with chronic obstructive pulmonary disease(COPD),and reveal the possible mechanism of the COPD treated with sodium houttuyfonate. Methods Twenty-four male Wistar rats were randomly divided into normal control group,model control group,dexamethasone group and sodium houttuyfonate group(n=6 for each). The rat models of COPD were established by intratracheal instillation of lipopolysaccharide and smudging. The expressions of PI3K and AKT1 and mTOR mRNA were determined by real-time PCR. The morphological changes of the lung tissue was examined by histopathology. Results Compared with the normal control group,the expressions of PI3K and AKT1 were significantly in-creased and mTOR mRNA was significantly decreased in the model group(P<0.01,P<0.05). Compared with the mod-el group,the expressions of PI3K and AKT1 were significantly decreased and mTOR mRNA was significantly increased in the sodium houttuyfonate group and dexamethasone group(P<0.01,P<0.05). Compared with the dexamethasone group, the expression of mTOR mRNA was significantly increased in the sodium houttuyfonate group(P<0.05). The pathological observation indicated that there were local pulmonary consolidation and a extensive neutrophil infiltration in the alveolar cav-ity. Prominent pulmonary interstitial fibrous hyperplasia was observed in the model group. The pathological manifestations were much ameliorated than those of the model group,and only mild interstitial pneumonia and a slight fibrous hyperplasia were seen in the sodium houttuyfonate and the dexamethasone groups. Conclusions Sodium houttuyfonate reduces the in-jury of lung tissue and has protective effect on COPD rats. The mechanism is probably related to the down-regulatation of expression of PI3K and AKT1 mRNA and up-regulatation of expression of mTOR mRNA in COPD rats.
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Objective:To investigate the association of AKT1 gene polymorphism with risperidone in the treatment of first-episode and untreated schizophrenia for 8 weeks.Methods:A total of 150 patients with Chinese schizophrenia who met DSM-Ⅳ,including 128 cases of risperidone (treatment dose 4-6 mg/d) for 8 weeks were treated with risperidone (treatment dose 4-6 mg/d).The Positive and Negative Symptom Scale (PANSS) reduction rate was used to evaluate the curative effect of drugs after 8 weeks.Using DNA sequencing,four single nucleotide polymorphisms (SNP) loci (rs1130214,rs10149779,rs1130233,rs2494732) genotype were detected in 128 Han patients with schizophrenia,and quantitative trait locus analysis (QTL) was used to explore the association between AKT1 gene polymorphisms and the efficacy of risperidone in the treatment of schizophrenia.Results:AKT1 gene rs1130233 (G ＞ A) and rs2494732(C ＞ T) were significantly associated with the increase in PANSS after 8-week risperidone treatment of schizophrenia (P ＜ 0.05).After repeated testing Bonferroni correction was still statistically significant.The correlation between rs1130214and rs10149779 in this sample was not statistically significant (P ＞0.05).Conclusion:This study suggests that polymorphisms of the AKT1 gene may be associated with the efficacy of risperidone in the treatment of schizophrenia in the Chinese Han population and is expected to provide a basis for the prediction of individual drug efficacy.
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Objective Effect of fluoxetine on the expression of angiogenic factors in hippocampus of depressive rats.Methods 32 male SD rats (3 month) of SPF level were randomly divided into fluoxetine group (n=8),fluoxetine ± LY294002 intervention group (n=8),model group (n=8) and control group (n =8).Control group without any treatment was free to eat and water for 5 weeks.The rats in the other groups were modeled by chronic,mild and unpredictable multiphase stress.The fluoxetine group was treated with fluoxetine for 2 weeks after modeling,and the intervention group was given LY294002 intervention,then the fluoxetine treatment for 2 weeks,the model group was free to eat and drink for 2 weeks.At the end of the 5th week,all the rats were decapitated and the hippocampus was removed on the ice bath.Western blot was performed to detect the expression of angiogenic factors.Results Compared with the model group (vascular endothelial growth factor(VEGF) (0.44 ± 0.08),fetal liver kinase 1 (FLK1) (0.37 ± 0.15),protein kinase B1 (AKT1) (0.40±0.10),p-AKT1 (0.53±0.07)),the expression of VEGF (0.98± 0.13),FLK1 (1.09± 0.21),AKT1 (1.05±0.05) and p-AKT1 (1.01±0.13)in hippocampus of fluoxetine group were significantly increased,and the differences were statistically significant (P＜ 0.01).And the expression of VEGT,FLK1,p-AKT1 in fluoxetine group increased compared with those of fluoxetine ± LY294002 intervention group (VEGF (0.55±0.08),FLK1 (0.55±0.14) and p-AKT1 (0.60±0.05)),and the difference were statistically significant (P＜0.01).Conclusion Fluoxetine can increase the expression of VEGF/FLK1 and p-AKT1 proteins in the hippocampus of depressed rats,which in turn may be involved in the regeneration of hippocampal blood vessels.This effect of fluoxetine may be related to the PI3K/AKT signaling pathway.
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Objective To detect the mRNA expression levels of PI3KCB and AKT1 genes in the peripheral blood lymphocytes of acute and chronic schizophrenia patients in different stages,and to explore the relationship between them and the clinic symptoms.Methods Twenty-four cases of schizophrenia patients without medication for at least 1 month,19 chronic schizophrenia patients with long-term clozapine medication,and 20 normal controls were involved in the study.The mRNA expression of PI3KCB and AKT1 genes of all the subjects were measured by real-time qRT-PCR,and the positive and negative symptom scales (PANSS) of schizophrenia patients were also evaluated.Spearman's correlation was used to analyze the relationship between PI3KCB,AKT1 and PANSS score.Results The gene expression of PI3KCB in acute schizophrenia patients,chronic schizophrenia patients with clozapine medication and normal control were (0.79±0.04),(0.83±0.08) and (0.87±0.09) respectively,and the difference was significant among the three groups (F=8.77,P=0.001).The AKT1 gene expression levels were (0.80±0.03),(0.27±0.13)and (0.29±0.12) respectively,and the difference was also significant among the three groups (F=302.31,P＜0.01).The PI3KCB mRNA levels of acute schizophrenia patients were significantly lower than the levels in healthy controls (MD =0.09,P=0.002),and the AKT1 mRNA levels of acute schizophrenia patients were significantly higher than the levels in chronic schizophrenia patients (MD=0.53,P＜0.01) and healthy controls (MD =0.51,P＜ 0.01).In schizophrenia patients,no significant relationship was found between PI3KCB,AKT1 expression levels and PANSS scores.Conclusion The gene expression status of PI3K-AKT pathway is significantly different in different stages of acute and chronic schizophrenia and that is no significant relationship with clinic symptoms,and clozapine treatment may affect its gene expression levels.
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Objective To investigate the effect and mechanism of miR-885-3p on the radiosensitivity of colorectal cancer cell HT-29. Methods The expression of miR-885-3p in HT-29 cells irradiated with different doses (0, 2, 4, 6, 8 Gy) of X-rays was detected by qPCR. The effect of miR-885-3p in modulating cell radiosensitivity was assessed in HT-29 cells with miR-885-3p overexpression. Bioinformatics prediction and dual luciferase reporter gene assay were employed to identify the direct target gene of miR-885-3p. Relationship between miR-885-3p and target gene tyrosine kinase 1 (AKT1) was investigated via regulation of miR-885-3p expression. The effect of AKT1 on radiosensitivity in HT-29 cells was evaluated through knockdown AKT1. The effect of AKT1 on miR-885-3p-induced radiosensitivity was detected by co-transferring miR-885-3p and AKT1 gene into HT-29 cells. Results miR-885-3p expression was up-regulated in radiation-induced HT-29 cells (F=46. 64, P<0. 05). Over-expression of miR-885-3p and knockdown of AKT1 enhanced cell radiosensitization by inhibiting survival and promoting apoptosis (t=12. 33, 12. 95, P <0. 05) with SER of 1. 602 and 1. 946, respectively. Inhibition of miR-885-3p promoted radioresistance by increasing cell survival and inhibiting apoptosis (t=11. 94, P<0. 05) with a SER of 0. 839. AKT1 is a target gene downstream of miR-885-3p, overexpression of AKT1 reversed the effect of miR-885-3p on cell radiosensitivity with a SER of 0. 680. Conclusions miR-885-3p can enhance the radiosensitivity of colorectal cancer HT-29 cells by directly targeting AKT1, which provides a target for improving the radiosensitivity of clinical colorectal cancer.
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Various posttranslational modifications (PTMs) participate in nearly all aspects of biological processes by regulating protein functions, and aberrant states of PTMs are frequently implicated in human diseases. Therefore, an integral resource of PTM-disease associations (PDAs) would be a great help for both academic research and clinical use. In this work, we reported PTMD, a well-curated database containing PTMs that are associated with human diseases. We manually collected 1950 known PDAs in 749 proteins for 23 types of PTMs and 275 types of diseases from the literature. Database analyses show that phosphorylation has the largest number of disease associations, whereas neurologic diseases have the largest number of PTM associations. We classified all known PDAs into six classes according to the PTM status in diseases and demonstrated that the upregulation and presence of PTM events account for a predominant proportion of disease-associated PTM events. By reconstructing a disease-gene network, we observed that breast cancers have the largest number of associated PTMs and AKT1 has the largest number of PTMs connected to diseases. Finally, the PTMD database was developed with detailed annotations and can be a useful resource for further analyzing the relations between PTMs and human diseases. PTMD is freely accessible at http://ptmd.biocuckoo.org.
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Humans , Databases, Protein , Disease , Genetics , Gene Regulatory Networks , Phosphorylation , Protein Processing, Post-Translational , Proteins , Metabolism , Search EngineABSTRACT
Objective To study the influence of O-GlcNAcylation on on proliferation and invasion of gastric cancer cells and evaluate the role of Aktl on O-GlcNAcylation promotting cells proliferation and invasion in gastric cancer.Methods Build the cell model:O-GlcNAc glycosylation levels rise or fall.The cell viability was determine by MTT.To investigate whether O-GlcNAcylation affected colony formation ability of gastric cancer cells,soft agar colony assays were carried out.Cell migration or invasion was using transwell chambers.The expression of Akt1 was detected through Western blot.Thiamet-G was used to eualuate the role of Akt1 on O-Gcnac cylation regulating invasion in gastric Cancei.Results O-GlcNAcylation was increased the gastric cancer cells proliferation ability,colony formation ability,migration and invasion ability in vitro.Akt1 was activated by Ser473 phosphorylation upregulation though O-GlcNAcylation.Akt1 shRNA was inhibition the cell invasive which induced by Thiamet-G.Akt1 overexpression was promoted by Thiamet-G-induced cell invasion.Conclusion O-GlcNAcylation enhanced oncogenic phenotypes possibly partially involving Akt1.