4982 Objective: 14-3-3 sigma, one isoform of the 14-3-3 family, plays a crucial role in the G2 checkpoint by sequestering Cdc2-cyclin B1 in the cytoplasm and the expression is frequently lost in human cancers. In this study, we examined the promoter methylation status and expression of 14-3-3 sigma, and evaluated its clinical significance in epithelial ovarian cancer. Materials and Methods: Twelve ovarian cancer cell lines, 2 ovarian surface epithelial cell lines, 8 normal, 8 benign, 12 borderline, and 102 ovarian cancer tissues were examined. Methylation specific PCR (MSP), quantitative RT-PCR, immunoblotting, and immunohistochemistry were employed to evaluate methylation status and expression of 14-3-3 sigma gene and protein. Results: Among the 12 ovarian cancer cell lines, the presence of methylated band was detected in 7 cell lines, 2 of which were together with unmethylated bands. Both of OSE cells were negative for methylated bands. By quantitative RT-PCR analysis, median values of relative 14-3-3 sigma gene expression in cancers with methylation (3.27) were significantly lower than those without methylation (16.4)(p 2cm, high serum CA125, high Ki-67 LI, and positive p53 immunoreactivity. In univariate analysis, 14-3-3 sigma immunoreactivity, age, performance status, grade, stage, residual tumor, and Ki-67 were significantly associated with overall survival. Negative 14-3-3 sigma cases were significantly worse overall survival and disease free survival (p=0.0058 and 0.002, respectively). In multivariate analysis, stage and residual tumor turned out to be independent prognostic factors. Conclusion: Our findings suggest that 14-3-3 sigma is inactivated mainly by aberrant DNA methylation and it may play an important role in the pathogenesis of epithelial ovarian cancer.
Supplementary Table 1 from Steroid Sulfatase and Estrogen Sulfotransferase in Colon Carcinoma: Regulators of Intratumoral Estrogen Concentrations and Potent Prognostic Factors
Supplementary Table 1 from Intratumoral Localization of Aromatase and Interaction between Stromal and Parenchymal Cells in the Non–Small Cell Lung Carcinoma Microenvironment
Supplementary Figures 1-2 from Intratumoral Localization of Aromatase and Interaction between Stromal and Parenchymal Cells in the Non–Small Cell Lung Carcinoma Microenvironment
Early growth responsive gene 3 (EGR3) is a zinc-finger transcription factor and plays important roles in cellular growth and differentiation. We recently demonstrated estrogen-mediated induction of EGR3 in breast carcinoma cells. However, EGR3 has not yet been examined in breast carcinoma tissues and its significance remains unknown. Therefore, in this study, we examined biological functions of EGR3 in the breast carcinoma by immunohistochemistry, in vitro study, and nude mouse xenograft model. EGR3 immunoreactivity was detected in carcinoma cells in 99 (52%) out of 190 breast carcinoma tissues and was associated with the mRNA level. EGR3 immunoreactivity was positively associated with lymph node status, distant metastasis into other organs, estrogen receptor alpha, or EGR3 immunoreactivity in asynchronous recurrent lesions in the same patients, and was negatively correlated with tubule formation. EGR3 immunoreactivity was significantly associated with an increased risk of recurrence and adverse clinical outcome by both uni- and multivariate analyses. Egr3-expressing transformant cell lines derived from MCF-7 Tet-Off cells (Eg-10 and Eg-11) significantly enhanced the migration and invasion properties according to the treatment of doxycyclin, but did not significantly change the cell proliferation. Moreover, Eg-11 cells injected into athymic mice irregularly invaded into the adjacent peritumoral tissues, although Clt-7, which was stably transfected with empty vector as a control, demonstrated a well-circumscribed tumor. Eg-11 cells were significantly associated with invasive components and less tubule formation in the xenograft model. These results suggest that EGR3 plays an important role in estrogen-meditated invasion and is an independent prognostic factor in breast carcinoma.
Abstract Nudix‐type motif 2 (NUDT2) hydrolyzes diadenosine 5′,5′′′‐p1,p4‐tetraphosphate (Ap4A) associated with various cellular functions. Previous studies demonstrated its regulation through estrogens, suggesting possible importance of NUDT2 in breast carcinoma. NUDT2, however, has not been examined in malignant tissues. Therefore, we examined its expression and functions in breast carcinoma. Immunohistochemistry for NUDT2 was examined by invasive ductal carcinoma (IDC: n = 145) and pure ductal carcinoma in situ (DCIS: n = 82), and NUDT2 mRNA was examined by real‐time PCR in 9 DCIS, 19 IDC and 6 non‐neoplastic breast tissues. We also used T47D breast carcinoma cells in in vitro studies. NUDT2 immunoreactivity was detected in 78% of DCIS and 63% of IDC, and NUDT2 mRNA level was significantly higher in DCIS or IDC than non‐neoplastic breast. NUDT2 status was significantly correlated with Van Nuys classification, HER2 or Ki‐67 in DCIS, and with stage, lymph node metastasis, histological grade or HER2 in IDC. NUDT2 status was significantly associated with adverse clinical outcome of IDC patients and proved an independent prognostic factor. Results of transfection experiments demonstrated that proliferation activity of T47D cells was significantly associated with NUDT2 expression level according to the treatment of estradiol and/or tamoxifen. NUDT2 expression was significantly decreased by estradiol, and it was also significantly decreased in T47D cells transfected with HER2 siRNA. These findings suggest that NUDT2 is an estrogen‐repressed gene and is also induced by HER2 pathways in breast carcinoma cells. NUDT2 promotes proliferation of breast carcinoma cells and is a potent prognostic factor in human breast carcinomas.
