ABSTRACT
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.
ABSTRACT
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.
ABSTRACT
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.