Expression of GATA3 in human breast cancer tissues and its clinical significance

AIM: To investigate the expression of GATA3 in human breast carcinoma and its clinical significance. METHODS: The expression level of GATA3 in breast cancer tissues from 124 patients was detected by the method of immunohistochemistry and the relationships between GATA3 expression and other clinicopathological factors were analyzed. RESULTS: Low expression of GATA3 in breast cancer tissues was associated with estrogen receptor (ER)/progesterone receptor (PR) negative, high histological tumor grade, p53 mutation and vascular invasion (P<005), but not with age, tumor size,human epidermal growth factor receptor 2 (HER-2) expression and lymph node metastasis (P>005). In all breast cancer tissues, the positive expression rate of GATA3 was 56.4%. The positive expression rate of GATA3 in luminal breast cancer is 684%, higher than that in non-luminal breast cancer (326%, P<005). In all breast cancer tissues, the expression of GATA3 in middle recurrence risk group was higher than that in high recurrence risk group (P<005). CONCLUSION: GATA3 expression in breast cancer is related to differentiation and biological characteristics of the tumor, which can be a factor for evaluation of the treatment and prognosis.     

Effect of Qingyi decoction on p-STAT3 expression in pancreas of mice with severe acute pancreatitis induced by L-arginine

AIM: To explore the role of STAT3 signaling pathway in acute pancreatitis induced by L-arginine, and the mechanisms of Qingyi decoction(QYD) treatment on severe acute pancreatitis (SAP). METHODS: The Kunming mice were randomly divided into 3 groups (n=10): control group, SAP group and QYD treatment group (SAP+QYD). The mice in SAP group and SAP+QYD group were intraperitoneally injected with 20% L-arginine (3 g/kg, bid). The mice in SAP+QYD group were also administered intragastrically with QYD(10 mL/kg) 30 min after the second injection of 20% L-arginine and twice a day for the following 2 days. The mice were anesthetized and sacrificed 72 h after SAP induction. The activity of amylase was measured in serum, the relative pancreatic weight was assayed, and the activity of myeloperoxidase (MPO) was analyzed to evaluate the neutrophil infiltration in lung tissues. The morphology of pancreas and lung was observed. The protein of pancreas was extracted to detect the expression of p-STAT3 by Western blotting. The mRNA expression of monocyte chemoattractant protein-1(MCP-1) was determined by real-time PCR. RESULTS: Compared with control group at 72 h, L-arginine induced SAP with increased serum amylase activity, pancreatic wet weight ratio and MPO activity in lung tissues (P<0.05). In SAP+QYD group, the activity of amylase, pancreatic wet weight ratio and MPO levels were significantly lower than those in SAP group (P<0.01). Compared with control group at 72 h, the pancreas and lung were obvious injured, the protein level of p-STAT3 and mRNA expression of MCP-1 in pancreas tissues increased significantly in SAP group. Compared with SAP group, the pathologic damage of the pancreas and lung tissues, the protein level of p-STAT3 and mRNA expression of MCP-1 in pancreas were significantly reduced in SAP+QYD group. CONCLUSION: The expression of p-STAT3 in pancreas increases in the mice with SAP induced by L-arginine. The activation of STAT3 may take part in the development of SAP. Inhibition of STAT3 activation in pancreas is one of the mechanisms of QYD treatment for SAP.     

Expression of KDM5B gene in breast cancer and its clinical significance

AIM:To investigate the expression of KDM5B gene in breast cancer tissues and its relationship with clinical data and prognosis of the patients. METHODS:Data sets of breast cancer were collected from The Cancer Genome Atlas (TCGA) database, and KDM5B mRNA expression profiles were downloaded. The mRNA expression of KDM5B in breast cancer tissues and adjacent tissues was detected by real-time PCR. The cases were divided into high expression group and low expression group according to the median expression of KDM5B, and the relationship with clinical data and case characteristics were analyzed. The relationship between KDM5B and prognosis of breast cancer patients was analyzed by Kaplan-Meier plotter. RESULTS:The expression of KDM5B in breast cancer tissues was significantly higher than that in normal breast tissues (P<0.01). In TCGA breast cancer data, the expression of KDM5B was significantly correlated with human epidermal growth factor receptor 2 (HER2), estrogen receptor (ER), age, histopathological type and lymph node metastasis (P<0.01), but not with progesterone receptor (PR), menopause and distant metastasis. The expression of KDM5B was significantly correlated with HER2, age and lymph node metastasis, but not with ER, PR, menopause, pathological type and distant metastasis. The higher the expression of KDM5B, the shorter the total survival time and the disease-free survival time of breast cancer patients. CONCLUSION:KDM5B is over-expressed in breast cancer tissues and correlated with prognosis of the patients. KDM5B expression is significantly correlated with HER2, age and lymph node metastasis. KDM5B may play an important role in the development of breast cancer.     

Expression of WNT5B in breast cancer and its clinical significance

AIM: To study the expression of WNT5B in the breast cancer and further to discuss the correlation between WNT5B and clinicopathologic characteristics of breast cancer. METHODS: The expression of WNT5B at mRNA and protein levels was measured by real-time PCR and Western blot in 67 cases of breast cancer and the tissue adjacent to carcinoma. In addition, the immunohistochemical method was used to detect the expression of WNT5B in the breast cancer and the tissue adjacent to carcinoma. The relationships between WNT5B expression and clinicopathologic indexes were also analyzed. RESULTS: The expression of WNT5B in the breast cancer was obviously lower than that in the tissue adjacent to carcinoma (P<0.05). The expression of WNT5B at mRNA and protein levels in 67 samples of breast cancer was in various degrees. The expression of WNT5B in T≤20 mm group of human breast cancer was obviously higher than that in T>20 mm group (P<0.05). The expression of WNT5B had no obvious correlation with axillary lymph node metastasis, histological grade and immunohistochemical indexes of ER, PR, c-ErBb-2, p53 and Ki67 (P>0.05) in the breast cancer. CONCLUSION: The expression of WNT5B decreases obviously in breast cancer. The expression of WNT5B is related to primary tumor size, which provides new ideas for the diagnosis and treatment of breast cancer, suggesting that WNT5B may be a new molecular marker for prognosis of breast cancer.     

Effect of heat shock protein 90 on proliferation of human breast cancer cells and its molecular mechanisms

AIM: To investigate the effect of heat shock protein 90 (HSP90) on proliferation of human breast cancer cells by a specific inhibitor for HSP90,geldanamycin (GA),and to analyze its molecular mechanisms.METHODS: Human breast cancer cell line MDA-MB-435s was used as a model.Proliferation of MDA-MB-435s cell was measured with MTT assay.Cell cycle was analyzed by flow cytometry.Alteration of phosphorylated Stat3 protein and mutant p53 protein in cells was detected by Western blotting.RESULTS: Geldanamycin inhibited the proliferation of MDA-MB-435s cells in time-and dose-depending manners.After treatment with GA,MDA-MB-435s cell cycle was arrested at G2/M phase.The levels of phosphorylated protein Stat3 and mutant p53 protein in MDA-MB-435s cells were reduced obviously by 55% and 48% respectively after 48 hour treatment with GA at the concentration of 400 nmol/L,compared to control cells.CONCLUSION: Inhibition of HSP90 function can prevent breast cancer cell proliferation significantly,which may be related to down-regulatin of the phosphorylated Stat3 protein and mutant p53 protein in the cells.     

Prognostic significance of FOXA1 and BRCA1 expression in triple negative breast cancer

AIM: To investigate the expression of forkhead box protein A1(FOXA1) BRCA1 protein,P53 and vascular endothelial growth factor (VEGF) in triple negative breast cancer (TNBC) and non-TNBC, and the relevance with the clinicopathological parameters for evaluating the prognosis. METHODS: The tumor samples were collected from 113 cases of breast cancer patients in the First Affiliated Hospital of Jinan University,and divided into TNBC group, luminal subtype group and HER-2 overexpression subtype group by the immunohistochemical results of estrogen receptor, progesterone receptor and HER-2. EnVision two-step method was used to detect the expression of FOXA1, BRCA1, P53 and VEGF in the tumor samples. RESULTS: Total FOXA1 positive expression rate was 63.7% (72/113), with 45.2% (19/42) in TNBC, 88.0% (44/50) in luminal subtype and 42.9% (9/21) in HER-2 overexpression subtype.The statistically sigfnificant difference among the 3 groups was observed (P<0.01). Total BRCA-1 positive expression rate was 47.8% (54/113), with 66.7% (28/42) in TNBC, 44.0% (22/50) in luminal subtype and 19.0% (4/21) in HER-2 overexpression subtype.The statisticallysignificant difference among the 3 groups was also observed (P<0.01). In the cases of clinical stages Ⅰ~Ⅱand histological grades 1~2, FOXA1 positive rate was higher than the FOXA1 negative rate (P<0.01). Negative correlations between FOXA1 positive rate and expression of P53/VEGF, and between FOXA1 positive rate and the recurrence rate were found (P<0.05). In the cases of clinical stages Ⅱ~Ⅲ and histological grades 2~3, the BRCA1 positive rate was higher than the BRCA1 negative rate (P<0.05). Positive correlations between BRCA-1 positive rate and the expression of P53/VEGF, and between BRCA1 positive rate and the recurrence rate were also observed (P<0.05). CONCLUSION: Expression of FOXA1 and BRCA1 in breast cancer is different. BRCA1 may be an adverse prognostic indicator for triple negative breast cancer.     

Expression of 2 SYK protein isoforms in breast cancer cells and their effect on cell invasion

AIM: This study was to explore the expression of spleen tyrosine kinase(SYK)(L) and SYK(S) in breast cancer cells and their effects on invasive phenotype of breast cancer cell line MB435. METHODS: The expression of SYK in breast cancer cell lines MB435, ZR75.1, MB468, T47D and MCF 7 were detected by Western blotting. MB435 cells were transfected with pcDNA3.1-SYK(L), pcDNA3.1-SYK(S) or pcDNA3.1 respectively, G418-resistant stable clones were pooled and the SYK expression was measured by Western blotting. Chemoinvasion assays were performed to study the effects of SYK(L) and SYK(S) on breast cancer cell invasion.RESULTS: SYK(L) and SYK(S) were simultaneously expressed in most detected breast cancer cells. Pooled SYK(L) and SYK(S) stable clones were made by Fugene 6 transfection and G418 selection. The expression of SYK(L) suppressed the invasive ability of breast cancer cells, while SYK(S) didnt.CONCLUSION: The simultaneously expression of SYK(L) and SYK(S) in breast cancer cells is a common phenomenon. Among SYK(L) and SYK(S),only SYK(L) protein is capable of suppressing the invasion of breast cancer cells.