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Objective To investigate the effects and mechanism of circTRIM33-12 on proliferation,apoptosis and epithelial-mesenchymal transition(EMT)of brain glioma cells by miR-191/DAB2 axis.Methods The expressions of circTRIM33-12,miR-191 and DAB2 in brain glioma cell CHG-5 and human normal brain glial epithelial cells HEB were detected by RT-PCR.The cultured CHG-5 cells were divided into the siRNA NC group,the circTRIM33-12 siRNA group,the DAB2 siRNA group;the mimics NC group,the miR-191 mimics group;the circTRIM33-12 WT+mimics NC group,the circTRIM33-12 WT+miR-191 mimics group,the circTRIM33-12 MUT+ mimics NC group,the circTRIM33-12 MUT+miR-191 mimics group;the inhibitor NC group,the miR-191 inhibitor group;the pcDNA+ mimics NC group,the pcDNA-TRIM33-12+mimics NC group,the pcDNA+miR-191 mimics group,the pcDNA-TRIM33-12+miR-191 mimics group;the DAB2 WT+mimics NC group,the DAB2 WT+miR-191mimis group,the DAB2 MUT+mimics NC group,the DAB2 MUT+ miR-191 mimis group.CCK-8 assay was used to detect the effects of the expressions of circTRIM33-12,miR-191 and DAB2 on the prolifera-tion ability of CHG-5 cells;flow cytometry was used to detect the effects of the expressions of circTRIM33-12,miR-191 and DAB2 on the apoptosis of CHG-5 cells;Western blot was used to detect the effects of the expressions of circTRIM33-12,miR-191 and DAB2 on EMT of CHG-5 cells.TargetScan database was used to analyze the correlations among miR-191,circTRIM33-12 and DAB2,and dual luciferase reporter gene assay was used to verify their relationships;RT-qPCR was used to detect the effect of circTRIM33-12 on DAB2 expression through miR-191.Results Compared with HEB cells,the expression of circTRIM33-12 in CHG-5 cells was down-regulated(P<0.01),the expression of miR-191 was up-regulated(P<0.01),and the expression of DAB2 was down-regulated(P<0.01).Compared with the siRNA NC group,the proliferation activity and N-cadherin expression of CHG-5 cells in the circTRIM33-12 siRNA group and the DAB2 siRNA group were significantly increased(P<0.01),while the apoptosis rate and E-cadherin expression were decreased(P<0.01).circTRIM33-12 targeted miR-191,and miR-191 targeted DAB2.Compared with the inhibitor NC group,the proliferation activity and N-cadherin expression of CHG-5 cells in the miR-191 inhibitor group were significantly decreased(P<0.01),while the apoptosis rate and E-cadherin expression were increased(P<0.01).circTRIM33-12 overexpression inhibited CHG-5 cell proliferation and EMT through miR-191.Conclusion circTRIM33-12 may regulate the proliferation,apoptosis and EMT of brain glioma cells through the miR-191/DAB2 axis.
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Purpose To investigate the clinical utility of DAB2IP (DOC-2/DAB2 interactive protein)and β-catenin expression in bladder urothelial carcinoma (BUC).Methods The expression of DAB2IP and β-catenin was detected in 104 BUC cases and 40 peritumorial tissues using EnVision two-step immunohistochemical method,and the association with BUC clinicopathological parameters was analyzed.Results The expression of DAB2IP in BUC was significantly less than that of peritumorial normal tissues,and the expression of β-catenin in BUS was significantly higher than that of peritumorial normal tissues (P ＜ 0.05).DAB2IP expression and histologic grading,clinical pathologic staging and 5 years survival rate had statistical significance (P ＜ 0.05).No statistical significance with gender,age,tumor diameter and in patients with incipient/recurrence.β-catenin expression and age,histologic grading,clinical pathologic staging,tumor diameter and 5 years survival rate have statistical significance (P ＜ 0.05).No statistical significance correlated with gender and in patients with incipient/ recurrence,DAB2IP and β-catenin expression in BUC are negatively correlated (P ＜ 0.05).Conclusion In bladder urothelial carcinoma,down-regulation of DAB2IP and up-regulation of β-catenin,are in a negative correlation.Abnormal expression of DAB2IP and β-catenin is correlated with histologic grading,clinical pathologic staging and prognosis.
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Objective To determine the expression and distribution of Disabled-2(Dab2) in normal human adrenal glands, and further to study the expression of Dab2 in tissues of different adrenocortical adenomas, and to elucidate whether Dab2 can be a specific molecular marker in the pathology of primary aldosteronism. Methods Real-time PCR and immunohistochemical staining were used to detect Dab2 expression in 10 aldosterone-producing adenoma (APA) samples, 8 cortisol-producing adenoma ( CPA) samples, 8 non-functioning adenoma ( NFA) samples and 6 normal adrenal samples. Results Immunohistochemical staining showed that Dab2 was significantly highly expressed in zona glomerulosa of normal human adrenal glands. Sporadical cluster of ZG cells with moderate Dab2 staining were demonstrated in APAs. In all CPA and NFA tumors, weak dab2 staining was detected. According to the results of real-time PCR, Dab2 mRNA expression was increased significantly in APAs compared with normal adrenal glands. There was no significant difference between normal adrenal glands, CPAs, and NFAs in regard to Dab2 mRNA expression. Compared to nontumor portions, APAs also showed higher Dab2 mRNA expression in the tumor( P<0. 05). Conclusion Dab2 was predominantly localized in zona glomerulosa in normal adrenal gland. Increased Dab2 mRNA expression was detected in APAs compared with normal adrenal glands. Whereas, Dab2 protein expression was just moderate increased in APAs. Weather Dab2 can be a specific molecular marker in the pathology of primary aldosteronism has to be further studied.
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Objective To construct recombinant lentiviral vector pSin-EF2-Puro-DAB2IP transfect prostate carcinoma PC3 cells,and to observe its transfection rate and expression level in PC3 cells.Methods The cDNA sequences specifically targeting the DAB2IP gene were designed and cloned into lentiviral vector PC3-pSin using DNA recombinant technique.Using PC3-pSin-EF2 as control,the 293T cells were transfected by Lipofectamine reagent for lentiviral particles packaged, and viral titer was determined. The DAB2IP mRNA and protein expression levels were examined by RT-PCR and Western blotting methods. Results The PCR and sequencing analysis results confirmed that the DAB2IP gene sequence was consistent with the sequence in GeneBank. The number of DAB2IP gene copy in PC3-pSin-EF2-DAB2IP cells and its control PC3-pSin-EF2 cells were 0.001±0.000 and 0.158 ± 0.013,respectively.Compared with control cells,the overexpression of mRNA in PC3-pSin-EF2-DAB2IP cells upregulated (179.370 ± 15.891)times,the difference was statistically significant (P<0.001). Compared with internal reference and control cells, the expression levels of DAB2IP protein in PC3-pSin-EF2-DAB2IP cells upregulated (2.431 ± 0.892)times and (2.415 ± 0.961)times respectively,the differences were statistically significant (P<0.001).Conclusion The lentiviral vector of the DAB2IP gene pSin-EF2-Puro-DAB2IP is successfully constructed,and its targeted gene is stably expressed in the targeted cells,which provides a basis for the further functional study of DAB2IP.
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The human Dab2 gene has a variety of physiological functions,and plays important roles in various signal transduction pathways.Researches have shown that reduced expression of Dab2 was closely related to the occurrence and development in many malignant cancers,and Dab2 has been regarded as a tumor suppressor.This paper reviewed about multiple researches on the structure and functions of Dab2,and explored the specific mechanisms in oncogenesis.
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Human DAB2 interaction protein (DAB2IP) is a novel member of Ras GTPase-activating protein family. It interacts directly with disabled-2 protein (DAB2/DOC2) which suppresses growth of cancers derived from different tissues, including mammary, prostate and ovarian cancers. DAB2IP was identified as an immediate downstream effector mediated by DAB2/DOC2. DAB2IP and DAB2/DOC2 form a unique protein complex that has a negative regulatory effect on the Ras-mediated signal pathway. It is demonstrated that DAB2IP is a tumor suppressor gene inactivated by methylation in several cancers. This article reviews the structure and biological functions of DAB2IP gene as well as its potential roles in carcinogenesis and evolution.
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Signal transduction is a very fine and complicated process that indudes participation in the cell propagation, division, maturation and apoptosis.Under normal circumstance disabled-2( Dab2) in human tissues participates in the signal transduction of the cell in order to allow the body to keep normal functions,but under some circumstances the absence of Dab2 influences the normal signal transduction path,leding to atypical cell changes.and the generation of tumors.This paper is a review on the construction and function of Dab2 and its related gene as well as the relation of Dab2 and tumors generation.