The far-upstream element-binding protein 2 is correlated with proliferation and doxorubicin resistance in human breast cancer cell lines.

Far-upstream element (FUSE)-binding protein 2 (FBP2) was a member of single-stranded DNA-binding protein family; it played an important role in regulating transcription and post-transcription and is involved in the regulation of C-MYC gene expression in liver tumors. However, the role of FBP2 in breast cancer and its mechanism has not been studied yet. Here, we discovered that FBP2 was up-regulated in breast cancer tissues and breast cancer cell lines. Moreover, immunohistochemistry analysis demonstrated that up-regulated FBP2 was highly associated with tumor grade, Ki-67, and poor prognosis, which was an independent prognostic factor for survival of breast cancer patients. At the cellular level, we found that FBP2 was correlated with cell cycle progression by accelerating G1/S transition, and knockdown of FBP2 could weaken cell proliferation, anchorage-independent cell growth, while enhancing the sensitivity of breast cancer cells to doxorubicin. More importantly, we found that activation of PI3K/AKT pathway could phosphorylate FBP2, and then make FBP2 shuttle from cytoplasm into the nucleus, which was the main mechanism of breast cancer cell proliferation and drug resistance. Taken together, our findings supported the notion that FBP2 might via PI3K/AKT pathway influence breast cancer progression and drug resistance, which might provide a new target for the design of anti-cancer drugs for breast cancer patients.

Decreased expression and prognostic role of cytoplasmic BRSK1 in human breast carcinoma: correlation with Jab1 stability and PI3K/Akt pathway.

OBJECTIVE: Jun activation domain-binding protein 1 (Jab1) was overexpressed in breast cancer, which was involved in degradation of the cyclin-dependent kinase inhibitor p27(Kip1). The objective of this study was to examine the effect of brain specific kinase 1 (BRSK1) expression on Jab1 over-expression and related signaling pathway in breast cancer. METHODS: Immunohistochemical analysis was performed in 95 human breast carcinoma samples and the data were correlated with clinicopathologic features. Furthermore, Western blot analysis was performed for BRSK1 and Jab1 in breast carcinoma samples and cell lines to evaluate their protein levels and molecular interaction. RESULTS: We found that the cytoplasmic BRSK1 expression was inversely associated with Jab1 expression (P<0.01) and correlated significantly with histologic grade (P=0.006), however nuclear BRSK1 expression couldn't obtain similar results. Kaplan-Meier analysis revealed that survival curves of low versus high expressers of cytoplasmic BRSK1 and Jab1 showed a highly significant separation in breast cancer (P<0.01). While in vitro, following release of breast cancer cell lines from serum starvation, the expression of Jab1, phosphor-Akt (p-Akt) was up-regulated, whereas BRSK1 and p27(Kip1) were decreased. Treatment of phosphatidylinositol 3-kinase (PI3K) inhibitor LY294002 could diminish Jab1 expression but increase BRSK1 expression. In addition, we employed siRNA technique to knock down Jab1 and/or BRSK1 expression and observed their effects on MDA-MB-231 cell growth. CONCLUSIONS: BRSK1 is a novel tumor suppressor in breast cancer which inversely correlated with Jab1 expression, may involve in the restoring Jab1-induced suppression of p27(Kip1) and may regulate cell cycle through the PI3K/Akt pathway.

Knocking down the expression of adenylate cyclase-associated protein 1 inhibits the proliferation and migration of breast cancer cells.

Adenylate cyclase-associated protein 1 (CAP1) is a conserved protein that was found to be up-regulated in breast cancer and related to the migration of breast cancer. We verified its roles in breast cancer specimens and cell lines. In our results, 71 of 100 specimens of breast cancer showed high levels of CAP1 by immunohistochemistry. Associated with statistical analysis, we saw that CAP1 was related to the grade of breast cancer. In MDA-MB-231, the expression of CAP1 was the highest and by knocking down the expression of CAP1 in MDA-MB-231, its ability for proliferating and migrating apparently decreased and induced changes in morphology, which were related to the arrangement of F-actin. Therefore, CAP1 might be a potential molecular targeted therapy for surgery and immune treatment.

[Expression and prognostic value of galectin-9 in hepatocellular carcinoma patients].

OBJECTIVE: To explore the prognostic value of galectin-9 in patients with hepatocellular carcinoma (HCC). METHODS: Galectin-9 was validated by immunohistochemisty in tissue microarrays from HCC patients (n = 147) and statistically assessed for the prognosis. The serum levels of galectin-9 from another independent cohort including HCC patients (n = 31) were assessed by enzyme-linked immunosorbent assay (ELISA). RESULTS: Patients with a lower expression of galectin-9 had significantly worse prognosis than those with a higher expression. The serum level of galectin-9 of HCC patients (8.36 ± 2.12) µg/L was significantly lower than that in healthy (4.62 ± 1.59 )µg/L and liver cirrhosis controls (5.11 ± 1.92 )µg/L (P < 0.05). Multivariate Cox proportional hazard analysis showed that galectin-9 was an independent marker for predicting a poor prognosis of HCC patients. CONCLUSION: Involved in the poor prognosis of HCC patients, galectin-9 may be a new predictor of recurrence for HCC patients and serve as a high-priority therapeutic target.

[Effects of adenoviral-mediated melanoma differentiation associated gene-7/IL-24 on growth and apoptosis of breast cancer cells].

OBJECTIVE: To investigate the effects of adenovirus melanoma differentiation associated gene-7 (mda-7)/IL-24 on the growth and apoptosis of human breast cancer cells. METHODS: Human breast cancer cells of the lines MCF-7 and MD-MBA-453 were cultured and transfected with purified adenovirus mda-7/IL-24. 48 h later, Western blotting was used to detect the protein expression of mda-7/IL-24 in the MCF-7 cells. The growth of Ad-mda-7/IL-24 in the MCF-7 cells was assayed by crystal violet staining and MTT method. The apoptotic effect of Ad-mda-7/IL-24 in the MCF-7 and MDA-MB-453 cells was detected by Hoechst staining and annexin V method. Nude mice were inoculated with MCF-7 cells and randomly divided into 2 groups to be injected with Ad-mda-7/IL-24 of PBS. The size of tumor was observed. RESULTS: Western blotting showed that mda-7/IL-24 was expressed in the transfected MCF-7 cells. Crystal violet staining and MTT method showed that Ad-mda-7/IL-24 dose and time-dependently inhibited the growth of the MCF-7 and MDA-MB-453 cells. Hochst staining and annexin V test indicated obvious apoptosis of the breast cancer cells. The size of tumor of the nude mice injected with Ad-mda-7/IL-24 was significantly smaller than that of the control mice. CONCLUSION: Ad-mda-7/IL-24 effectively inhibits the growth of breast cancer cells and induces its apoptosis.

[Expression and significance of c-jun-activation-domain binding protein (JAB1) in hepatocellular carcinoma].

OBJECTIVE: to investigate the expression of c-jun-activation-domain binding protein (JAB1) in hepatocellular carcinoma (HCC) and the clinicopathologic significance thereof. METHODS: Immunohistochemistry was used on 76 specimens of HCC tissues and native liver tissues adjacent to the HCC tissues, and 10 specimens of normal liver tissues near the liver angioma to detect the expression of JAB1 and the cell proliferative factor Ki67. RESULTS: The expression rate of JAB1 in the HHC tissues was 68.85%, significantly higher than that in the adjacent liver tissues and normal liver tissues near liver angioma (38.72% and 34.36% respectively, both P < 0.001). The expression of JAB1 was correlated with the histological differentiation, serum alpha-fetoprotein level, and metastasis (P = 0.000, 0.015, and 0.000), but not with gender, age, tumor size, serum HBsAg, and existence of necrosis and cirrhosis. The expression rate of Ki67 protein in the HCC tissues was 41.45%, significantly higher than that in the adjacent liver tissues and normal liver tissues near liver angioma (2.11% and 2.01% respectively, both P < 0.001). There was a positive correlation between JAB1 and Ki67 (P < 0.001). CONCLUSION: JAB1 is highly expressed in HCC and may play an important role in the oncogenesis and development of HCC. JAB1 may be used as a marker for neoplastic change in liver cells and thus has potential clinicopathologic value.

Aldehyde dehydrogenase 1 expression is correlated with poor prognosis in breast cancer.

Breast cancer (BC) is one of the most common cancers worldwide, and is a major cause of death in women. Aldehyde dehydrogenase 1 (ALDH1) is a marker of stem cells and cancer stem cells, and its activity correlates with the outcome of various tumors, including BC. This study aimed to analyze the relationship between ALDH1 expression and clinicopathological characters in BC and the prognostic significance of ALDH1.We used quantitative reverse-transcription PCR (qRT-PCR) to detect ALDHA1 mRNA levels in 25 fresh frozen BC samples and matched noncancerous samples. Immunohistochemistry on tissue microarrays was used to analyze protein expression in 137 paraffin-embedded BC tissues and corresponding noncancerous tissues. STATA 16.0 software was used for statistical analysis.The results suggested that levels of both ALDH1 mRNA and protein in BC were significantly higher than in corresponding adjacent breast samples (3.856 ±â€Š0.3442 vs 1.385 ±â€Š0.1534, P < .001; 52.6% vs 25.5%, P < .001, respectively). ALDH1 protein expression was also significantly associated with histological grade (P  =  .017), tumor size (P  =  .017), and tumor-node-metastasis (TNM) stage (P  =  .038). Multivariate analysis using the Cox regression model demonstrated that ALDH1 expression (P  =  .024), molecular typing (P  =  .046), and TNM classification (P  =  .034) were independent predictive factors for the outcome of BC. Kaplan-Meier analysis and the log-rank test indicated that patients with high ALDH1 expression, triple-negative BC, and advanced TNM stage had a reduced overall survival time.These data suggest that ALDH1 could be used as a prognostic factor for BC and may provide a useful therapeutic target in the treatment of BC.

Overexpression of SYF2 promotes cell proliferation and correlates with poor prognosis in human breast cancer.

SYF2, a known cell cycle regulator, is reported to be involved in cell cycle arrest by interacting with cyclin-D-type binding protein 1. In the present study, we investigated the role of SYF2 in human breast cancer (BC) progression. SYF2 was highly upregulated in BC tissues and cell lines, as per Western blot and immunohistochemistry analysis. The SYF2 expression level had a significant correlation with the tumor grade and Ki-67 expression. In vitro starvation-refeeding experiment and SYF2-siRNA transfection assay demonstrated that SYF2 could promote proliferation of BC cells, while SYF2 knockdown resulted in cells cycle arrest at G1/S phase, reducing the cell growth rate of BC cells. These results indicated that SYF2 promotes human BC progression by accelerating the BC cells' proliferation. SYF2 could be a novel therapeutic target in human BC therapies.

[Clinical research on angiogenesis in colorectal carcinoma and expression of CK20 mRNA in peripheral blood].

OBJECTIVE: To investigate the correlation between microvessel density (MVD) and expression of vascular endothelial growth factor (VEGF) in tissues of colorectal carcinoma and the micrometastasis of tumor cells in these patients' peripheral blood. METHODS: The MVD and expression of VEGF were evaluated immunohistochemically while the micrometastasis of tumor cells in these patients' peripheral blood was detected by RT-PCR method. RESULTS: The average count of MVD in high and middle differentiation grade was 30.2 +/- 12.7, while in low differentiation grade 86.6 +/- 19.1. The expression of VEGF was positive in 26 patients (44.8%). The MVD and positive expression of VEGF were correlated to differentiations. stage and metastasis of colorectal carcinoma. CK(20) mRNA was found in peripheral blood of 32 patients (55.2%) and the positive rate was up to 60.4% 48 hours after operation, among which positive rate in the radical resection group was 47.7% and in the non-radical resection group 85.7%. 11 out of 21 patients positive in CK(20) mRNA turned to negative 7-14 d after radical resection, while 11 out of 12 patients remained positive at the same time after non-radical resection. The expression of CK(20) mRNA was correlated to the stage and metastasis of the cancer. The MVD and positive expression of VEGF were higher in patients with positive expression of CK(20) mRNA. CONCLUSIONS: The MVD and positive expression of VEGF were correlated to differentiation, stage and metastasis of colorectal carcinoma. The angiogenesis in tissues of colorectal carcinoma was closely related to the micrometastasis of tumor cells in these patients' peripheral blood. The detecting of CK(20)mRNA by RT-PCR may be a sensitive method for evaluating the micrometastasis colorectal carcinoma in peripheral blood and help in prognosis prediction, effect assessment and guidance of multipurpose therapy.