Carcinoma, Hepatocellular ; metabolism ; pathology ; Core Binding Factor Alpha 3 Subunit ; metabolism ; Female ; Hepatocytes ; metabolism ; Humans ; Liver ; metabolism ; pathology ; Liver Neoplasms ; metabolism ; pathology ; Male ; RNA, Messenger ; genetics

Adolescent ; Child ; Child, Preschool ; Core Binding Factor Alpha 3 Subunit ; genetics ; metabolism ; DNA Methylation ; Female ; Humans ; Infant ; Lymphoma ; genetics ; metabolism ; Male ; Promoter Regions, Genetic

OBJECTIVETo investigate the expression of Runt-related transcription factor gene 3(RUNX3) in gastric cancer and its impact on the outcome of gastric cancer patients.

METHODSBy using immunohistochemistry staining and western blot assay, the expression of RUNX3 protein in 66 cases of gastric cancer with various clinicopathologic characteristics were detected, and the effects of RUNX3 protein expression on the outcome of patients undergone surgical resection were evaluated.

RESULTS(1) The expression rate of RUNX3 protein in gastric cancer lesions was 60.6% (40/66), and RUNX3 protein was mainly expressed in the cytoplasm of cancer cells. RUNX3 protein expression in tumor tissues was significantly higher than that in non-tumor tissues. (2) RUNX3 protein expression was correlated with tumor differentiation (P=0.025) and Lauren's classification (P=0.034), but had no relationship with the TNM stage (P=0.085). (3) In sharp contrast, the median survival time of patients who had tumors with negative and positive RUNX3 protein expression were 2478 and 2187 days respectively (P=0.016).

CONCLUSIONSRUNX3 protein influences the differentiation of gastric cancer. The role of RUNX3 protein as a tumor-suppressor in tumorigenesis and differentiation of gastric carcinoma need to be further evaluated.

Aged ; Cell Differentiation ; Core Binding Factor Alpha 3 Subunit ; genetics ; metabolism ; Female ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Neoplasm Staging ; Prognosis ; Stomach Neoplasms ; genetics ; pathology

OBJECTIVETo analyze the relationships among the aberrant methylation of Runx3 gene promoter, the Runx3 protein expression and clinicopathological features in gastric cancer.

METHODSMethylation specific PCR was used to measure the promoter methylation status of Runx3 gene in tumor and the adjacent normal mucosal tissues from 40 patients with gastric cancer. Protein expression of Runx3 was measured by immunohistochemistry.

RESULTSThe frequency of promoter methylation of Runx3 gene in gastric cancer tissue(55.0%) was significantly higher compared to the adjacent normal tissues (12.5%)(P<0.01). The positive rate of protein expression of Runx3 in gastric cancer tissue(37.5%) was significantly lower compared to the adjacent normal tissues (100%). There was marked association between hypermethylation and negative protein expression (P<0.05). The frequency of Runx3 promoter methylation was associated with histological type, N grade, and tumor stage.

CONCLUSIONThe promoter hypermethylation is a main mechanism of reduced or loss expression of Runx3 gene, which may provide molecular diagnosis and stage evaluation of gastric cancer.

Adult ; Aged ; Core Binding Factor Alpha 3 Subunit ; genetics ; metabolism ; CpG Islands ; DNA Methylation ; Female ; Humans ; Middle Aged ; Neoplasm Staging ; Promoter Regions, Genetic ; Stomach Neoplasms ; diagnosis ; metabolism ; pathology

OBJECTIVETo set up a method of analyzing gene expression profile from mouse whole embryos.

METHODSMouse whole mount RNA in situ hybridization(WM-ISH) of E10.5-E14 embryos was carried out by using digoxigenin-labeled Runx1 and Runx3 RNA probes and their expression profile was observed by detecting the existence and status of corresponding mRNAs in the embryonic tissues.

RESULTSClear hybridization signals were observed in different tissues and organs hybridized by Runx1 or Runx3 RNA probe. Different probes and ages of embryos had need of their own optimal proteinase K treatment conditions.

CONCLUSIONMouse whole mount RNA in situ hybridization is an effective method of analyzing gene expression. It is useful for revealing whole gene expression profile and has a great potentiality in the era of functional genomics. It provides an alternative method of studies on gene expression which is at least as good as LacZ staining and immunohistochemistry. The key factor of the success to mouse whole mount RNA in situ hybridization is whether the proteinase K treatment conditions are optimal or not.

Animals ; Core Binding Factor Alpha 2 Subunit ; Core Binding Factor Alpha 3 Subunit ; DNA-Binding Proteins ; genetics ; Embryo, Mammalian ; metabolism ; Gene Expression Profiling ; methods ; Gene Expression Regulation, Developmental ; In Situ Hybridization ; methods ; Mice ; Proto-Oncogene Proteins ; genetics ; RNA ; genetics ; metabolism ; RNA, Messenger ; genetics ; metabolism ; Sensitivity and Specificity ; Transcription Factors ; genetics

OBJECTIVE To explore the role of runt-related transcription factor 3(RUNX3) in the tumorgenesis and progression of cervical carcinoma. METHODS The immunohistochemical staining technique was used to detect the expression of RUNX3 protein in 25 cases of normal cervix, 34 intraepithelia neoplasia (CIN), and 48 cervical carcinomas. SYBR Green I chimeric fluorescence Real-time PCR was applied to detect the expression of RUNX3 mRNA in 10 cases of normal cervix, 24 CIN, and 30 cervical carcinomas. RESULTS The expressions of RUNX3 protein and mRNA in normal cervix, CINI,CINII-III, and cervical carcinoma tissues tended to be down-regulated. There was significant difference among these groups (P<0.05). The expressions of RUNX3 protein and mRNA in the cervical carcinoma tissues were correlated with the histological differentiation, clinical stage, and lymphatic metastasis (P<0.05), but had no relationship with the age, high-risk human papillomavirus infection, and histological classification (P> 0.05). CONCLUSION RUNX3 may function as a tumor suppressor gene in the occurrence and progression of cervical carcinoma.
Adult ; Carcinoma, Squamous Cell ; genetics ; metabolism ; Cervical Intraepithelial Neoplasia ; genetics ; metabolism ; Core Binding Factor Alpha 3 Subunit ; genetics ; metabolism ; Disease Progression ; Female ; Humans ; Middle Aged ; RNA, Messenger ; genetics ; metabolism ; Uterine Cervical Neoplasms ; genetics ; metabolism

OBJECTIVETo investigate the relationship between hypermethylation of Runx3 gene promoter and estrogen receptor (ER) and the implications of Runx3 gene promoter hypermethylation in ER positive breast cancer.

METHODSWestern blot and RT-PCR were used to detect the protein and mRNA expression of Runx3 gene in breast cancer cell lines (MCF7 and SKBR3) and normal breast epithelium cell line (MCF10A). Immunohistochemical SP method was used to detect the expression of ER and Runx3 proteins in 113 tissue samples of breast cancer. Moreover, methylation specific PCR was used to detect RUN3 promoter methylation in cell lines MCF7, SKBR3, MCF10A and 113 tissue samples of breast cancer.

RESULTSOf the 3 cell lines, Runx3 protein and mRNA were detectable in MCF10A, but were absent in MCF7 and SKBR3. MCF7 had a high methylation status at Runx3 promoter, in contrast, MCF10A and SKBR3 showed unmethylated RUN3 promoter. Among the 113 cases of breast cancer, 68 cases were ER positive and 45 were negative. The positive rates of Runx3 protein expression in ER positive and negative tumors were 26.5% (18/68) and 66.7% (30/45), respectively (P<0.05). Runx3 promoter hypermethylation was seen in 82.4% (56/68) of ER positive breast cancer cases and 22.2% (10/45) of ER negative ones (P<0.05). Among 68 cases of ER positive cases, Runx3 promoter hypermethylation was positively correlated with the clinical tumor stage (OR=5.84, P<0.05).

CONCLUSIONSRunx3 gene promoter hypermethylation is present mainly in the ER positive breast cancers. Testing of Runx3 promoter methylation may provide additional reference for clinical stage and prognosis of breast cancer patients, especially in those with ER positive tumors.

Breast Neoplasms ; genetics ; metabolism ; Cell Line, Tumor ; Core Binding Factor Alpha 3 Subunit ; genetics ; metabolism ; DNA Methylation ; Female ; Humans ; Neoplasm Proteins ; genetics ; metabolism ; Polymerase Chain Reaction ; methods ; Prognosis ; Promoter Regions, Genetic ; RNA, Messenger ; metabolism ; Receptors, Estrogen ; genetics ; metabolism

OBJECTIVETo investigate the evolution pattern of the Runx3 gene 5'-CpG island ~3478 bp region methylation in human salivary gland adenoid cystic carcinoma (SGACC).

METHODSQuantitative MSP method was used to detect the methylation status of CpG island in various regions (No.1-10) of Runx3 promoter region, and Western blot was used for detection of the expression of Runx3 protein in 41 salivary gland SGACC samples and corresponding non-neoplastic salivary gland tissues. A Logistic model was used to analyze the risk ratio between the methylation status of CpG island in Runx3 gene and development of salivary SGACC, meanwhile, the possible association among the methylation of Runx3 gene, the clinicopathological parameters of SGACCs, and Runx3 protein expression was compared.

RESULTSThe results of qMSP showed that the hypermethylation initially occurred at the most 5' region of the Runx3 CpG island and spread to the transcription start site. The methylation rate was highest in region No. 1 and No. 2 among the successive ten regions ranging from the 5' region to the transcription start site within the Runx3 CpG island, and lowest in the transcription start site both in SGACCs and normal salivary glands. Furthermore, there was no methylation in the transcription start site in nomal salivary glands tissues. Together with the results of Logistic model analysis, those results indicate that the transcription start site within the Runx3 promoter CpG island is critical for gene silencing. Western blot results revealed that the Runx3 protein level in SGACC was significantly lower than that in normal salivary glands (P < 0.01). In combination of the results of qMSP, it is presumed that the Runx3 gene methylation is one of the reason inducing the down-regulation of Runx3 in SGACCs.

CONCLUSIONSMethylation of the Runx3 CpG island spreads from the most 5'-region to the transcription start site in human salivary gland adenoid cystic carcinoma, and the transcription start site may be a critical region for the methylation of Runx3. The evolution pattern of Runx3 gene methylation is related to the tumorigenesis of SGACCs.

Adult ; Aged ; Aged, 80 and over ; Carcinoma, Adenoid Cystic ; genetics ; metabolism ; pathology ; Core Binding Factor Alpha 3 Subunit ; genetics ; metabolism ; CpG Islands ; genetics ; DNA Methylation ; Female ; Humans ; Logistic Models ; Male ; Middle Aged ; Salivary Gland Neoplasms ; genetics ; metabolism ; pathology ; Salivary Glands ; metabolism

OBJECTIVETo investigate the expression of RUNX3 protein in hepatic cell carcinoma (HCC) and its relationship with the clinicopathological factors of HCC.

METHODSImmunohistochemistry was performed to detect the expression of RUNX3 protein in HCC and the surrounding normal tissues, and the relation of RUNX3 with the clinicopathological factors of HCC was analyzed.

RESULTSThe positivity rate of RUNX3 protein expression was 49.02% (25/51) in HCC tissues, significantly lower than that in the surrounding normal tissues [82.35% (42/51), Χ(2)=12.5706, P<0.01). RUNX3 protein expression varied significantly with such pathological factors as the differentiation degree, cancer-associated thrombosis in the portal vein and intrahepatic metastasis (P<0.05), but not with tumor diameter, location, tumoral hemorrhage, necrosis or histotypes of the tumor (P>0.05).

CONCLUSIONRUNX3 protein expression is lowered in HCC as compared with that in the surrounding normal tissue, suggesting an important role of RUNX3 in the tumorigenesis and development of HCC and the possible identity of RUNX3 gene as an anti-oncogene of HCC.

Carcinoma, Hepatocellular ; genetics ; metabolism ; pathology ; Core Binding Factor Alpha 3 Subunit ; genetics ; metabolism ; Down-Regulation ; Female ; Genes, Tumor Suppressor ; Humans ; Liver Neoplasms ; genetics ; metabolism ; pathology ; Male ; Tumor Suppressor Proteins ; genetics ; metabolism

OBJECTIVE: To explore the relationships between hypermethylation of human runt-related transcription factor 3 (Runx3) gene promoter and laryngeal squamous cell cancer. METHOD: Promoter hypermethylation and mRNA expression were detected by methylation-specific PCR and RT-PCR. RESULT: The expression of Runx3 gene mRNA detected in laryngeal carcinoma (1.62 +/- 1.01) was lower than that in adjacent tissues samples (5.66 +/- 2.07) (t = 10.72, P < 0.01). No methylation of Runx3 promoter was found in adjacent tissues samples. But hypermethylation was found in 95.0% (38/40) of the laryngeal carcinoma specimens. The rate of methylation of Runx3 promoter in laryngeal carcinoma was higher than that in adjacent tissues (P < 0.01). The Runx3 mRNA were down-regulated in lymphnode metastasis or poorly differentiated groups, but the Runx3 promoter methylation were detected in those groups markedly. CONCLUSION Hypermethylation of Runx3 promoter is one of the inactivation re-seasons in laryngeal squamous cell carcinoma, and the decreasing of Runx3 mRNA expression may be related to lymph node metastasis and development of laryngeal squamous cell carcinoma.
Adult ; Aged ; Carcinoma, Squamous Cell ; genetics ; metabolism ; pathology ; Core Binding Factor Alpha 3 Subunit ; genetics ; metabolism ; DNA Methylation ; Female ; Gene Expression Regulation, Neoplastic ; Humans ; Laryngeal Neoplasms ; genetics ; metabolism ; pathology ; Lymphatic Metastasis ; Male ; Middle Aged ; RNA, Messenger ; genetics