Prognostic significance of CUEDC2 expression in hepatocellular carcinoma

AIM: To investigate the expression of CUE domain-containing 2 (CUEDC2) in hepatocellular carcinoma (HCC) and to analyze its clinical prognostic significance. METHODS: Total 186 formalin-fixed paraffin-embedded tissues obtained from surgical HCC with detailed clinicopathological and follow-up data were used. The expression of CUEDC2 was detected by immunohistochemistry. The relationships between the expression of CUEDC2 and clinicopathological characteristics and prognosis were analyzed. RESULTS: The positive rate of CUEDC2 in HCC was 85.5% (159/186), among which, the low expression was 52.2% (97/186) and the high expression was 47.8% (89/186). CUEDC2 expression was correlated with serum alpha-fetal protein (AFP) level, tumor size, tumor number, tumor differentiation and TNM stage (P<0.05). Kaplan-Meier survival curves showed that the patients with high expression of CUEDC2 were associated with significantly shorter overall survival and recurrence-free survival than those with low CUEDC2 expression (P<0.05). Multivariate Cox regression analysis revealed 3 independent prognostic factors including CUEDC2 expression, serum AFP and tumor number. CONCLUSION: CUEDC2 was expressed in most HCC tissues, which was relevant to tumor growth, tumor differentiation and prognosis. CUEDC2 could be a novel valuable molecular marker to predict the HCC prognosis.     

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.     

ACTL8 expression and its correlation with clinicopathological features and prognosis in breast cancer

AIM: To investigate the actin-like protein 8 (ACTL8) expression and its relationship with clinicopathological features and prognosis in breast cancer.METHODS: The expression of ACTL8 in human normal mammary epithelial cell line MCF-10A and 5 breast cancer cell lines was detected by Western blot. The expression of ACTL8 was also investigated by immunohistochemistry in 6 cases of breast cancer specimens with adjacent normal tissues. The data in 488 cases of breast specimens from TCGA dataset were downloaded, and the relationship between the mRNA expression of ACTL8 and the clinicopathological features and prognosis was analyzed.RESULTS: The expression of ACTL8 in 4 breast cancer cell lines was significantly higher than that in breast epithelial cell line MCF-10A.The level of the ACTL8 expression in breast tumors was significantly higher than that in the corresponding adjacent normal breast tissues. The mRNA expression of ACTL8 was correlated with age, tumor size, clinical TNM stage and lymph node metastasis of breast cancer patients (P < 0.05). The high expression level of ACTL8 mRNA indicated a poor prognosis of breast cancer patients. CONCLUSION: ACTL8 protein is highly expressed in breast cancer specimens and is closely correlated with the clinicopathological features and prognosis, suggesting that ACTL8 is a prognostic marker for breast cancer or a potential new target for treatment of breast cancer.     

Changes of MDR1 and MRP gene expression in primary breast cancer after neoadjuvant chemotherapy

AIM: To investigate the expression of drug resistance genes, MDR1 and MRP, in patients with primary breast cancer after neoadjuvant chemotherapy. METHODS: MDR1 and MRP gene expression were detected by semi-quantitative RT-PCR in 20 patients with primary breast cancer before and after chemotherapy. RESULTS: Before chemotherapy, MDR1 and MRPexpression could be detected in 15 cases (75%) and 18 cases (90%), respectively. After chemotherapy, expression of MDR1 was not significantly different from that before chemotherapy, but expression of MRPwas significantly different from that before chemotherapy. CONCLUSION: Drug resistance gene MRP, but not MDR1 expression is enhanced in patients with primary breast cancer subjected to neoadjuvant chemotherapy.     

Expression of ALEX1 in breast cancer and its clinical significance

AIM: To detect the expression of ALEX1 in the breast cancer tissues in order to verify whether ALEX1 has correlation with clinical pathological features in breast cancer.METHODS: Real-time PCR and immmunohistochemistry were applied to detect the expression of ALEX1 at mRNA and protein levels in the breast tissues. The statistical analysis were performed for determining the correlation with the level of ALEX1 and the clinical pathological features in breast cancer.RESULTS: The protein levels of ALEX1 in the breast cancer tissues were lower than that in the non-breast cancer tissues (P<0.01). The expression of ALEX1 had correlations with pathological grade, clinical stage, molecular type (P<0.05) but had no correlation with the patients' age, tumor size and tumor types in breast cancer. Furthermore, the result of real-time PCR showed that mRNA expression of ALEX1 was also significantly reduced in the breast cancer tissues (P<0.01).CONCLUSION: The expression of ALEX1 in the breast cancer tissues is lower than that in non-breast cancer tissues. The pathological grade and clinical stage in breast cancer are negatively correlated with the expression of ALEX1.     

Abnormal expression and clinical significance of RASA1 in pancreatic carcinoma

AIM: To investigate the role of Ras p21 protein activator 1 (RASA1) in the development and progress of pancreatic cancer. METHODS: Different expression levels of RASA1 were measured by qRT-PCR and Western blotting in the pancreatic cancer cells Capan-2, CFPAC-1 and BxPC-3, and the pancreatic ductal cell line H6C7. Besides, the different expression levels between the pancreatic cancer and the pancreatic benign lesions, such as chronic pancreatitis or pancreatic cyst, were detected by the method of immunohistochemistry. The relationship between the clinicopathological feature and the RASA1 expression was analyzed. RESULTS: Both mRNA and protein expression levels of RASA1 decreased in pancreatic cancer cells compared with the ones in the pancreatic ductal cells (P<0.05). The protein level of RASA1 in the pancreatic cancer tissues was lower than that in the pancreatic benign lesion tissues (P<0.05). The pancreatic cancer samples with adjacent organ invasion had a significantly lower expression level of RASA1 than that in the pancreatic cancer samples limited in the pancreas (P<0.05). The expression levels of RASA1 were much higher in the cancers on stage I than the ones on stage II or Ⅲ (P<0.05). However, no relationship between the RASA1 expression level and the maximum diameter of cancer, the lymph node invasion and the survival time was observed. CONCLUSION: RASA1 plays an important role in the pancreatic cancer development as a potential tumor suppressor.     

Relationship between c-erbB-2, BCSG1 expression and recurrence,metastasis in breast cancer

AIM: To assess the significance of c-erbB-2, BCSG1 (breast cancer specific gene-1) expression and other parameters in recurrence or metastasis of breast cancer. METHODS: The expression of c-erbB-2, BCSG1, and ER, PR, MVD, VEGF, VEGF-C, FLT-4, LVD were determined with the SP immunohistochemical method in 58 cases of invasive breast cancer patients occurred over 5 years. The cases were used to analyze the effect of c-erbB-2, BCSG1, VEGF-C and ER, PR, MVD, VEGF, FLT-4, LVD expression on clinical-pathological manifestations and prognosis in breast cancer. RESULTS: The expression rates of c-erbB-2, BCSG1, VEGF-C, LVD were respectively 25.9%, 62.1%, 36.2%, 32.8% in association with the lymph node metastasis and recurrence of breast cancer (P<0.05), the expression rate of MVD was also increased significantly (P＜0.05). CONCLUSION: The c-erbB-2, BCSG1, VEGF-C, LVD are highly expressed and strongly correlated with the lymph node metastasis and recurrence of breast cancer, of which BCSG1 may be used as a predictor of prognosis.     

Expression and prognostic significance of SLP-2 in gastric cancer

AIM: To investigate the expression of the red cell membrane integration protein SLP-2 (stomatin-like protein 2) in gastric cancer tissues and to analyze its correlation with clinicopathological manifestations and prognosis. METHODS: One hundred and ninety gastric cancer tissue samples with detailed clinical information were collected from the Department of Pathology, Sun Yat-sen University Cancer Center. The protein expression of SLP-2 in ganstric cancer was detected by the method of immunohistochemistry. The relationships between SLP-2 expression and the clinicopathological manifestations were evaluated. RESULTS: The positive rate of SLP-2 in gastric cancer tissue was 63.2% (120/190). SLP-2 expression was relevant to infiltration depth, TNM stage and lymph node metastasis (P<0.05). However, no statistical difference was observed in the SLP-2 expression associated with sex, age, differentiation, tumor size and distant metastases. Kaplan-Meier survival curves revealed that increased expression of SLP-2 was associated with poor prognosis in gastric adenocarcinoma patients (P<0.01). Based on the univariate analysis, 7 factors were found to have statistical significance of associations with overall survival, including SLP-2 expression, lymph node metastasis, histological grade, tumor size, invasive depth, distant metastases and the 7th edition of the UICC TNM classification. Only the tumor size and the 7th edition of the UICC TNM classification were independent prognostic factors for overall survival in the multivariate analysis. CONCLUSION: SLP-2 is highly expressed in gastric adenocarcinoma tissues and may play an important role in tumor progression and metastasis. Although SLP-2 is not an independent prognostic factor, it may influence the prognosis of gastric cancer. Increased expression of SLP-2 can be used for predicting unfavorable prognosis in gastric adenocarcinoma patients.     

Effect of curcumin on expression of p21 and CD44V6 in human breast cancer xenografts

AIM:To study the effect of curcumin on the expression of p21 and CD44V₆ in breast carcinoma in nude mice.METHODS:Nude mice were xenografted with human breast cancer cell line MCF-7 and randomly divided into 2 groups (n=4 in each group): control group and curcumin group. In latent period,the percentage of tumor development was observed. Tumors were measured and the surface areas were calculated. RT-PCR was performed to detect the expression level of cyclin D1,p21 and CD44V₆ mRNA. RESULTS:The tumor surface areas in the curcumin group were significantly lower than those in control group. In curcumin treatment group,the expression of p21 was up-regulated while cyclin D1 was nearly not changed. The expression of CD44V₆ was significantly down-regulated in curcumin group.CONCLUSION:Curcumin inhibits the expression of CD44V₆ and up-regulates the expression of p21 in nude mice bearing human breast cancer cell line MCF-7.     

FoxM1 promotes development of AML by regulating Bcl-2 expression

AIM: To investigate the role of Forkhead box M1 (FoxM1) and B-cell leukemia/lymphoma-2 (Bcl-2) in the pathogenesis of acute myeloid leukemia (AML). METHODS: RT-qPCR and immunofluorescence analysis were used to determine the expression of FoxM1 at mRNA and protein levels in AML-de novo patients, AML-complete remission (CR) patients, AML-refractoriness and relapse (RR) patients and healthy controls. HL60 cells and K562 cells were transfected with FoxM1 siRNA. The cell proliferation was detected by cell proliferation assay and colony formation assay on soft agar, and the cell apoptosis was determined by flow cytometry. The expression of FoxM1 and Bcl-2 at mRNA and protein levels was detected by RT-qPCR and Western blotting. The activity of bcl-2 promoter was examined by luciferase reporter assay with FoxM1 targetting. RESULTS: FoxM1 expression level in the AML-de novo patients was significantly higher than that in the healthy controls. As compared with the AML-de novo patients, FoxM1 expression in the AML-CR patients was reduced, and the FoxM1 expression level was the highest in the AML-RR patients. FoxM1 expression was inhibited in the HL60 cells and K562 cells transfected with FoxM1 siRNA. Transfection with FoxM1 siRNA in the HL60 cells and K562 cells inhibited the proliferation as compared with NC siRNA transfection, and impaired the colony formation ability. On the contrary, transfection with FoxM1 siRNA promoted the cell apoptosis. FoxM1 regulated bcl-2 expression positively. CONCLUSION: FoxM1 promotes the development of AML by regulating bcl-2 expression. Silencing of FoxM1 expression suppresses cell proliferation and promotes cell apoptosis. FoxM1 is a potential target for AML treatment.     

Relationship between expression of somatostatin and DNA ploidy in breast cancer

AIM: To investigate the relationship between somatostatin and the pathologic type, estrogen receptor,DNA ploidy of nuclei in tumor cells of breast cancer.METHODS: 67 cases of primary breast cancer and 25 cases of benign breast tumor were examined by immunohistochemical stretomyces avidin peroxidase method. 26 cases of breast cancer selected at random were analyzed by flow cytometry.RESULTS: Somatostatin expressed significantly higher in low malignant breast cancer than that in high malignant breast cancer (P<0.05). Most of cancers with positive staining of somatostatin were diploidy,most of cancers with negative staining were aneuploidy,there had significant difference between two groups (P<0.05).CONCLUSION: Somatostatin may delay the progress of breast cancer,and somatostatin levels in cancer tissues may become a useful indicator for assessing prognosis of patients with breast cancer.     

Expression and clinical significance of Bmi-1 in gastric carcinoma

AIM: To investigate the relationship between expression of Bmi-1 (B cell-specific MLV integration site-1) in gastric cancer and its clinicopathologic significance.METHODS: 146 surgical patients with gastric carcinoma were followed up at least 2 years.Expression of Bmi-1 protein was examined by immunohistochemistry in their archival paraffin embedded tissue specimens.RESULTS: The intensive positive rate of Bmi-1 expression in gastric cancer was 67.8% (99/146).Expression of Bmi-1 was highly correlated with tumor size,clinical stage,lymph node metastasis and T classification (P<0.05),but not with sex,age,tumor differentiation,etc.(P>0.05).The survival rate in the patients with Bmi-1 expression was much lower than that in those patients without Bmi-1 expression (P<0.01).Multivariate analysis indicated that Bmi-1 expression,T classification,lymph node metastasis,distant metastasis,tumor size and postoperative chemotherapy were all significantly prognostic factors of gastric carcinoma.CONCLUSION: Overexpression of Bmi-1 in patients with gastric carcinoma enhances the possibility of invasion and metastasis,implying a poor prognosis.Bmi-1 may serve as fairly a good prognostic factor to indicate biologic behavior and prognosis in gastric carcinoma.     

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.     

Silencing of FoxM1 induces apoptosis of oral squamous-cell carcinoma cells by promoting mitochondrial release of cytochrome C

AIM: To study the effect of forkhead box protein M1 (FoxM1) silencing on apoptosis of oral squamous-cell carcinoma. METHODS: Oral squamous-cell carcinoma SCC9 cells were infected with FoxM1-shRNA lentivirus or negative control lentivirus. The silencing effect was measured by RT-qPCR and Western blot. The changes of cell viability effect was measured by MTT saay. The cell colony formation ability was measured by plate experiment. Flow cytometry was used to analyze the changes of apoptotic rate. Western blot was used to measure the protein levels of cleaved caspase-3 and cleaved caspase-9 in the cells. The changes of mitochondrial membrane potential were measured by JC-1 method. Western blot was used to measure the protein level of cytochrome C in the mitochondria and cytoplasm. RESULTS: Infection with FoxM1-shRNA lentivirus successfully reduced the expression of FoxM1 in oral squamous-cell carcinoma cells (P<0.05). Negative control lentivirus had no effect on the expression level of FoxM1 in the cells. The cell viability was reduced by FoxM1 silencing, and the ability of cell colony formation was also decreased. The apoptotic rate and the protein levels of cleaved caspase-3 and cleaved caspase-9 were all increased (P<0.05), and the mitochondrial membrane potential was decreased. The protein level of cytochrome C in the cytoplasm was increased, while the protein level of cytochrome C in the mitochondria was decreased (P<0.05). CONCLUSION: Silencing of FoxM1 induces the apoptosis of oral squamous-cell carcinoma cells by decreasing the mitochondrial membrane potential and promoting the release of cytochrome C from mitochondria.     

Inhibition of FoxM1 sensitizes leukemia K562 cells to homoharringtonine

AIM: To study whether inhibition of forkhead box protein M1(FoxM1) sensitizes leukemia K562 cells to homoharringtonine (HHT). METHODS: K562 cells were incubated with HHT at different concentrations (0μmol/L, 0.015μmol/L, 0.030μmol/L and 0.045μmol/L) for different time (0 h, 24 h, 48 h and 72 h). The mRNA and protein levels of FoxM1 were detected by real-time PCR and Western blot. FoxM1 siRNA was transfected into K562 cells with 0.015μmol/L HHT after 6 h. After 72 h incubation, the cell proliferation was detected by cell counting and soft agar assay, and the proportion of apoptotic K562 cells was determined by flow cytometry. The expression of c-Myc and Sp1 were detected by real-time PCR and Western blot. RESULTS: FoxM1 expression was reduced time-dependently and dose-dependently, suggesting that HHT mediated the downregulation of FoxM1 in K562 cells. In K562 cells, treatment with FoxM1 siRNA and HHT inhibited the cell proliferation and promoted the apoptosis significantly. Therefore, inhibition of FoxM1 sensitized leukemia K562 cells to HHT. The expression of c-Myc and Sp1 was positively regulated by FoxM1. CONCLUSION: HHT inhibits Forkhead box protein M1 expression in K562 cells. Inhibition of FoxM1 sensitizes K562 cells to HHT.     

Relationship between zinc and zinc transporter expression in breast cancer MCF-7 cells

AIM: To study the changes of zinc transporter gene expression in MCF-7 cell line exposed to ZnCl₂ and TPEN. METHODS: Human breast cancer cell line MCF-7 was exposed to different concentrations of ZnCl₂ (0, 50, 100, 150, 200 μmol/L) and TPEN (0, 5, 10, 15 μmol/L), respectively. Twelve hours later, the cell viability was measured by MTT and levels of zinc transporter mRNA by RT-PCR. Zinquin was used to estimate the intracellular zinc concentrations. RESULTS: MCF-7 cells viability rate was significantly decreased when exposed to ZnCl₂ (with 150 μmol/L and 200 μmol/L) and TPEN. The intracellular zinc concentration was significantly increased when exposed to ZnCl₂ and decreased when exposed to TPEN. ZnT-1 mRNA level was increased along with the increasing concentration of ZnCl₂ but decreased when exposed to TPEN. The expressions of ZIP2 and ZIP10 were increased along with the increasing concentration of TPEN. CONCLUSION: ZnT-1 gene expression is induced by zinc supplement and repressed by zinc deficiency. ZIP2 and ZIP10 gene expressions are induced by zinc deficiency.     

Mechanism of microRNA-138-5p inhibiting proliferation,migration and invasion of lung cancer cells

AIM: To investigate the mechanism of microRNA-138-5p (miR-138-5p) inhibiting the proliferation, migration and invasion abilities of lung cancer cells.METHODS: The lung cancer A549 and H460 cells were transfected with miR-NC (control group) or miR-138-5p (experimental group). The bioinformatic analysis was performed to predict the target genes of miR-138-5p.The expression levels of miR-138-5p, forkhead box protein C1 (FOXC1) mRNA and vimentin mRNA were detected by RT-qPCR. The protein expression of FOXC1, vimentin, E-cadherin, N-cadherin and β-catenin was determined by Western blot. MTS method and colony formation assay were used to detect cell viability and proliferation ability. Wound healing assay and Transwell assay were used to detect cell migration and invasion ability.RESULTS: Over-expression of miR-138-5p significantly reduced the expression of FOXC1 and vimentin at mRNA and protein levels (P<0.05). The expression of E-cadherin and β-catenin were up-regulated and the expression of N-cadherin was down-regulated. The proliferation, migration and invasion abilities of the lung cancer cells were inhibited by the over-expression of miR-138-5p.CONCLUSION: miR-138-5p inhibits the proliferation, migration and invasion abilities of lung cancer cells by targeting FOXC1 and vimentin. It may be a potential target for lung cancer gene therapy.     

Correlation between E-cadherin and FOXO3a expression in gastric can-cer tissues and cells

AIM: To investigate the expression of E-cadherin and forkhead box protein O3a (FOXO3a) in gastric cancer tissues and cells, and its correlation with cell viability. METHODS: The expression of E-cadherin and FOXO3a was detected by immunohistochemical staining in 53 specimens of gastric cancer tissues and their adjacent tissues, and the relationship between their expression and clinicopathological characteristics were analyzed. E-cadherin-over-expressing gastric cancer AGS cells were constructed by lentivirus-mediated cell transfection, and the protein expression of E-cadherin and FOXO3a was detected by immunocytochemistry method. The expression of E-cadherin, FOXO3a, Akt, Bcl-2 and Bax was determined by Western blot. The cell viability was detected by CCK-8 assay. RESULTS: The positive expression rates of E-cadherin and FOXO3a proteins in gastric cancer tissues were both significantly lower than those in their adjacent tissues (P<0.05). E-cadherin positive expression in gastric cancer tissues was significantly related to tumor grade and TNM stage (P<0.05), but not related to age, sex, location, T stage or lymph node metastasis. FOXO3a positive expression was significantly related to tumor grade (P<0.05), but not related to age, sex, location, TNM stage, T stage or lymph node metastasis. The expression of E-cadherin was positively correlated with FOXO3a expression in gastric cancer tissues (r=0.376, P=0.003). After over-expression of E-cadherin, the viability of gastric cancer AGS cells was significantly inhibited, the expression of FOXO3a, Bcl-2 and Bax was significantly increased, and the expression of Akt was significantly decreased. CONCLUSION: E-cadherin and FOXO3a are involved in the development of gastric cancer, and E-cadherin may affect the viability of gastric cancer cells by regulating Akt/FOXO3a signaling pathway.     

Relationship between apoptosis,proliferation and expression,mutation of related genes in breast cancer

AIM:To study the relationship between apoptosis, proliferation and expression,mutation of related genes in breast cancer.METHODS:Methods of TUNEL, immunohistochemical S-P and PCR-SSCP were used respectively to study apoptotic index (AI), mitotic index(MI), expression of Bcl-2,p53,c-erbB-2,PCNA,Ki67,TopoⅡ and mutation of p53 in 54 cases of breast cancer.RESULTS:AI and MI were 9.40±3.78 and 5.96±2.36, respectively. There was a significant direct correlation between them(r=0.46.P＜0.01).High expression of Bcl-2,PCNA,Ki67,TopoⅡ coincided with high AI,MI(P＜0.01). High expression of p53,c-erbB-2 and mutation of p53 coincided with high MI(P＜0.01). Type of p53 mutation coincided with AI(P＜0.05).CONCLUSION:Disturbance of gene control between apoptosis and proliferation is related with expression,mutation of related genes in breast cancer.