Methylation of TIMP-3 gene and their relationships with their clinical-pathological characteristics of colorectal cancers

AIM: To investigate whether methylation of the TIMP-3 gene is associated with clinical-pathological characteristics, recurrence and metastasis of the colorectal cancer. METHODS: Nest methylation specific PCR (nMSP) and RT-PCR techniques were used to detect methylation of TIMP-3 gene and its mRNA expression in the colorectal cancer specimen and adjacent non-cancerous tissues. RESULTS: The expression of TIMP-3 mRNA in tumor tissues was distinctly reduced (P<0.01). The expression of TIMP-3 mRNA in those without lymph node metastasis was higher than those with lymph node metastasis (P<0.01). The patients with Duke's C, D and lymph node metastasis were more to contain methylated TIMP-3 compared to those with Duke's A, B and no lymph node metastasis (P<0.05). Statistical differences in pathological characteristics such as tumor site, Duke's stage, histological differentiation and type between TIMP-3 methylation positive group and negative group were observed (P<0.05). CONCLUSION: Methylation of the TIMP-3 gene promoter usually occurs in the proximal site, infiltrating type, poor cellular differentiation, lymph node metastasis and advanced stage of colorectal cancers patients.     

Hypermethylation of SFRP genes in esophageal squamous cell cancer

AIM: To investigate the promoter methylation of secreted frizzled-related protein(SFRP) genes in esophageal squamous cell cancer(ESCC). METHODS: The methods of methylation-specific PCR(MS-PCR) and RT-PCR were applied to examine the CpG methylation of the SFRP promoter and the mRNA expression of SFRP genes,respectively, in 78 samples of ESCC and corresponding adjacent non-cancer tissues. The protein expression of β-catenin was determined by immunohistochemistry.RESULTS: In the ESCC tissues, the frequencies of promoter methylation in SFRP1 , SFRP2 , SFRP4 and SFRP5 genes were 65.4%(51/78), 69.2%(54/78), 62.8%(49/78) and 52.6%(41/78),respectively, significantly higher than those in the adjacent tissues(P<0.01). The hypermethylation of these genes had no correlation with clinical stage and pathological classification in ESCC tissues(P>0.05). The frequency of simultaneous methylation of the 4 genes was correlated with the clinical stage(P<0.05). The positive rates of mRNA expression of the 4 genes in ESCC tissues were 42.3%(33/78), 46.2%(36/78), 50.0%(39/78) and 39.7%(31/78), respectively lower than those in the adjacent tissues(P<0.01). The mRNA expression of SFRP genes and the ectopic expression of β-catenin were correlated with the methylation frequency of SFRP genes(P<0.01).CONCLUSION: Promoter methylation of SFRP1 , SFRP2 , SFRP4 and SFRP5 genes was a frequent event in ESCC, indicating a contribution to the pathogenesis of ESCC through aberrant canonical Wnt/β-catenin signaling pathway. Combination analysis of methylation status in SFRP genes may has definite value on estimating prognosis of ESCC.     

Expression and clinical significance of RhoC and Ki-67 in human esophageal squamous cell carcinoma tissues

AIM: To investigate the expression and significance of RhoC and Ki-67 in human esophageal squamous cell carcinoma (ESCC) tissues.METHODS: The expression of RhoC and Ki-67 was detected in 52 specimens of ESCC by the method of immunohistochemistry. The clinicopathological features were also analyzed.RESULTS: The expression of RhoC was detected in 32 of the total 52 (61.5%) cases of human ESCC tissues, significantly higher than that in the adjacent histologically normal epithelium, which was only in 11 of 37 cases (29.7%, P<0.05). RhoC expression was closely correlated with clinical tumor-node-metastasis (TNM) stage (P<0.05) and lymph node metastasis (P<0.05) in ESCC. The over-expression of RhoC was positively correlated with Ki-67 in ESCC (r=0.322, P<0.05).CONCLUSION: The over-expression of RhoC protein significantly correlates with advanced TNM stage, lymph node metastasis and cell proliferation ability of ESCC. Therefore, RhoC may be a new auxiliary parameter for early diagnosis and prognostic evaluation of ESCC.     

《中国蔬菜品种志》

本书由中国农业科学院蔬菜花卉研究所主编,已于2002年9月出版发行。全书分上、下卷,1～6章为上卷,包括根菜类、白菜类、芥菜类、甘蓝类、绿叶菜类及葱蒜类,计2263个品种,1347页;7～12章为下卷,包括瓜类、茄果类、豆类、薯芋类、水生蔬菜类和多年生蔬菜类,计2550个品种,1177页。入志的品种中,地方品种占     

Expression and significance of Mnk2 and eIF4E in esophageal squamous cell carcinoma

AIM: To investigate the expression and significance of MAPK-interacting kinase-2 (Mnk2) and eukaryotic initiation factor 4E (eIF4E) in the patients with resected esophageal squamous cell carcinoma (ESCC).METHODS: The protein expression of Mnk2 and eIF4E in ESCC tissues (98 cases) and normal esophageal tissues (20 cases) were assessed by immunohistochemistry (IHC), and their correlations with clinicopathological features were statistically analyzed.RESULTS: The over-expression rate of Mnk2 and eIF4E was 68.4% (67/98) and 61.2% (60/98), respectively. The expression of Mnk2 had a positive correlation with eIF4E (P<0.05). Clinicopathologic analysis showed that Mnk2 expression was significantly correlated with T classification (P<0.05) and clinical stage (P<0.05).CONCLUSION: The over-expression of Mnk2 was significantly related to the tumor invasive depth, TNM stages and expression of eIF4E in ESCC. Expression of Mnk2 and eIF4E may have a cooperative formation mechanism in the development of ESCC.     

Clinical significance of STMN1 expression in cervical cancer and effect of inhibition of its expression on viability and apoptosis of cervical cancer SiHa cells

AIM: To investigate the clinical significance of stathmin 1 (STMN1) expression in cervical cancer and the influence of its expression on the viability and apoptosis of cervical cancer cells. METHODS: Western blot was used to detect the protein expression of STMN1 in cervical cancer tissues, and the relationship between the expression and clinical characteristics of cervical cancer was analyzed. STMN1-siRNA was transfected into cervical squamous-cell carcinoma SiHa cells. The protein levels of STMN1, STAT3, p-STAT3 and survivin were determined by Western blot after transfection for 48 h. The cell viability was measured by MTT assay. The apoptosis was analyzed by flow cytometry. DCFH-DA probe was used to detect the level of reactive oxygen species (ROS). RESULTS: The protein expression of STMN1 in cervical cancer tissues was significantly higher than that in paracancerous tissues (P<0.01). The STMN1 protein expression level was not correlated with age and histological types of cervical cancer patients, but was related to clinical stage, histological differentiation and lymph node metastasis (P<0.01). Transfection with STMN1-siRNA significantly reduced the expression of STMN1 in SiHa cells. Compared with control group, the cell viability in STMN1-siRNA group was significantly decreased, the apoptotic rate and ROS content were increased, and the protein levels of p-STAT3 and survivin were down-regulated (P<0.01). However, no significant difference of the STAT3 protein level was observed between STMN1-siRNA group and control group. CONCLUSION: STMN1 is highly expressed in cervical cancer, and its expression is related to clinical stage, histological differentiation and lymph node metastasis. Inhibition of STMN1 expression reduces the viability and promotes apoptosis of cancer cells by down-regulating STAT3 signaling pathway.     

Expression and significance of T-cell immunoglobulin and ITIM domain in colorectal cancer

AIM: To study the expression and clinical significance of T-cell immunoglobulin and ITIM domain (TIGIT) in colorectal cancer. METHODS: The patients with colorectal cancer (n=80) from January 2016 to June 2018 were selected. The expression of TIGIT and CD155 in the colorectal cancer tissues and adjacent normal tissues were detected by immunohistochemical staining method. The expression of TIGIT and CD155 was also determined by Western blot and ELISA. RESULTS: The positive expression rates of TIGIT and CD155 were 78.8% (63/80) and 83.8% (67/80) in the colorectal cancer tissues, significantly higher than that in the paracancerous tissues of 8.8% (7/80) and 18.8% (15/80), respectively (P<0.05). There was a positive correlation between TIGIT and CD155 expression (r=0.867, P<0.01). The expression levels of TIGIT and CD155 were increased as the stage evolved. The positive rates of TIGIT and CD155 in the colorectal cancer tissues were correlated with the degree of differentiation, pathological stage and lymph node metastasis (P<0.05). CONCLUSION: TIGIT and CD155 are correlated with the occurrence and development of colorectal cancer, and can be used as one of the prognostic indicators of colorectal cancer.     

Protein and mRNA expression of KAI1 metastasis suppressor gene and its clinical significance in cervical carcinoma

AIM: To study the mRNA and protein expression of Kang ai1 (KAI1) tumor suppressor gene and to determine the relationship between KAI1 and invasiveness and metastasis of cervical cancer. METHODS: The expression of KAI1 metastasis suppressor was detected by immunohistochemistry in paraffin slides and by real-time quantitative polymerase chain reaction (RT-PCR) in fresh tissue. The samples included 20 cases of normal cervical tissues, 20 cases of cervical intraepithelial neoplasia (CIN) and 40 cases of cervical carcinoma. The results of the gene expression combined with the pathological and clinical data were also analyzed. RESULTS: The expression of KAI1 protein and mRNA was related to the tissue differentiation of cervix. The positive rates of KAI1 expression were the highest in the normal cervical tissue, the middle in CIN and the lowest in cervical carcinoma with significant difference among three groups (P<0.01). The expression of KAI1 protein was not related with the grade of CIN (P>0.05). However, both mRNA and protein expression of KAI1 were related to the differentiation and the clinical stages of cervical cancer (P<0.01) and also related to the metastasis of the cancer. The positive rates between the non-lymphatic metastasis and lymphatic metastasis (P<0.05) were significant different. Cox regression and logistic regression showed that the tissue differentiation, clinical stages, lymphatic metastasis and expression of KAI1 were all related factors with recurrence and prognosis of cervical cancer. CONCLUSION: The down-regulation of KAI1 tumor suppressor gene at both mRNA and protein levels is related to the differentiation, clinical stages and metastasis of cervical cancer, indicating that the expression of KAI1 is a prognostic factor for cervical cancer.     

Expression and clinical significance of bone morphogenetic protein 3 in hilar cholangiocarcinoma tissues

AIM: To investigate the expression and clinical significance of bone morphogenetic protein 3 (BMP3) in hilar cholangiocarcinoma tissues.METHODS: Thirty cases of hilar cholangiocarcinoma specimens were collected. The expression of BMP3 at mRNA and protein levels in the tumor tissues and paracancerous tissues was detected by real-time PCR and Western blot. The hilar cholangiocarcinoma paraffin-embedded specimens (n=103) were collected. The protein expression of BMP3 was determined by immunohistochemical method, and the relationship of BMP3 protein expression with clinical pathological characteristics was evaluated.RESULTS: In the 30 patients with hilar cholangiocarcinoma, the expressions of BMP3 protein and mRNA in 22 cases of tumor tissues were significantly decreased compared with the adjacent normal tissues. The results of immunohistochemistry showed that 87 cases were negative and 16 cases were weakly positive in all 103 cases of hilar cholangiocarcinoma. The expression of BMP3 protein was associated with the tumor TNM staging, lymph node metastasis and tumor differentiation (P<0.05).CONCLUSION: BMP3 gene might be inhibited in human hilar cholangiocarcinoma. The down-regulation of BMP3 gene might be associated with the carcinogenesis and development of hilar cholangiocarcinoma.     

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.     

Correlation between Mnk2 protein expression and prognosis of patients with esophageal squamous cell carcinoma

AIM: To investigate the protein expression of mitogen-activated protein kinase-interacting kinase-2 (Mnk2) and its prognostic effect in the patients with resected esophageal squamous cell carcinoma (ESCC). METHODS: A total of 86 informative patients with surgically resected ESCC and 54 normal esophageal tissues were enrolled. Western blot and immunohistochemistry (IHC) were utilized to assess the protein expression of Mnk2, and its correlation with prognosis was statistically analyzed by the methods of Kaplan-Meier curve and Cox proportional hazard mode. RESULTS: The protein expression of Mnk2 was elevated in most of tumor tissues compared with the adjacent tissues. Clinicopathologic analysis showed that Mnk2 expression was significantly correlated with the TNM stage (P<0.05). Both disease-free survival (DFS) and overall survival (OS) of Mnk2 over-expression patients were shorter than those in Mnk2 negative expression group. Multivariate analysis confirmed that Mnk2 expression, as an independent and significant factor for both DFS and OS, predicted a poor prognosis of the patients with resected ESCC (P<0.05). CONCLUSION: The expression of Mnk2 was significantly related to the TNM stages, and might be a novel predictor for prognosis in ESCC.     

Relationship between genetic instability of nm23H1 gene and clinical pathological characteristic in hepatocellular carcinoma

AIM: The aim of this study was to examine the microstatellite instability (MSI) and loss of heterozygosity(LOH) of locus D17S396 on chromosome 17 and their influence on the expression of nm23H1 in hepatocellular carcinoma (HCC),which may provide experimental evidence for the mechanism of nm23H1 gene and tumor metastasis.METHODS: Techniques such as DNA extraction from formalin-fixed paraffin-embedded tissues,PCR-SSCP,ordinary silver stain were used to study MSI and LOH of locus D17S396.Envision immunohistochemistry and Leica-Qwin computer imaging techniques were used to assess the expression of nm23H1.RESULTS: ① The frequency of heredity instability of HCCs was 35.42%.The frequency of LOH in the cases with lymph node or distant organs metastasis or not and with intrahepatic metastasis or embolus of portal vein or not was significantly different (P<0.01),it was higher in stage TNM Ⅲ than that in stageⅠ and Ⅱ.Moreover,it was higher in high tendency to invasion or metastasis cases than that in the low tendency cases (P<0.01).② The expression of nm23H1 was 56.25%.It was significantly different in Edmondson grade,TNM stage and in lymph node or distant organ metastasis cases (P<0.01).The cases with high tendency of invasion or metastasis exhibited lower nm23H1 expression compared with low tendency cases (P<0.01).③ The positive rate of nm23H1 protein in LOH positive group was lower than that in LOH negative group (P<0.05).CONCLUSION: The results indicate that both MSI and LOH of nm23H1 gene control the development of HCC independently in different pathways.LOH inhibits the expression of nm23H1,which endows it with high aggressive and poor prognosis.Increase in the amount of nm23H1 protein expression effectively restrains the tendency to invasion or metastasis of HCCs and improves prognosis of patients.     

Expression of Wnt/β-catenin signaling pathway-related proteins in breast cancer and its clinical significance

AIM: To evaluate the expression of Wnt/β-catenin signaling pathway-related proteins in breast cancer and the significance. METHODS: The patients with breast cancer (n=150) in our hospital from January 2015 to January 2017 were selected as study object. The tumor tissue samples of these patients were obtained from paraffin section of breast cancer by surgical resection with complete clinicopathological data. The corresponding paracancerous tissue sam-ples were taken from the non-tumor tissue samples from the above breast cancer patients, which were 0.5~1 cm away from the tumor tissue. The methods of real-time PCR and Western blot were performed to examine the expression of Wnt-1 and β-catenin at mRNA and protein levels. Human breat cancer MCF-7 cells were divided into 3 groups:control group (MCF-7 cells without treatment), agonist group[MCF-7 cells+Wnt3a (1 mg/L)] and antagonit group[MCF-7 cells+DKK1 (16 μmol/L)]. The expression of Wnt-1 and β-catenin at mRNA and protein levels was detected by real-time PCR and Western blot. RESULTS: Compared with the paracancerous tissues, the expression levels of Wnt-1 and β-catenin were higher in tumor tissues at mRNA and proteins levels (P<0.05). Notably, the positive expression rates of Wnt-1 and β-catenin were significantly higher in tumor tissues than that in the paracancerous tissues. Furthermore, Wnt-1 expression was associated with tumor metastasis (χ²=5.352, P=0.021), tumor stage (χ²=9.412, P=0.002) and tumor size (χ²=9.412, P=0.002). In addition, β-catenin expression was also associated with tumor metastasis (χ²=9.851, P=0.002) and tumor stage (χ²=5.661, P=0.017). Compared with control group, the expression of Wnt-1 and β-catenin at mRNA and protein levels in agonist group was increased (P<0.05),while that in antagonist group was decreased (P<0.05). CONCLUSION: The expression levels of Wnt-1 and β-catenin related with Wnt/β-catenin signaling pathway are increased in the breast cancer, which are closely related to the malignant state of the tumor.     

Methylation status of multiple genes in colorectal cancer tissues and its clinical significance

AIM: The aim of the present study was to detect the methylation status of multiple genes (Syk, CDH1 and TIMP-3) in colorectal cancer (CRC) tissues, to investigate the roles in the pathogenesis of CRC, to investigate whether methylation of multiple genes was associated with recurrence and metastasis of CRC, and to evaluate its ability to predict the prognosis of CRC. METHODS: Nested methylation-specific PCR (n-MSP) were used to detect methylation status of spleen tyrosine kinase (Syk) from 120 CRC tissues and the methylation status of CDH1 and tissue inhibitor of metalloproteinase-3 (TIMP-3) from 100 CRC tissues. RESULTS: We found that methylation in one or more genes, co-methylation in two genes and three genes were detected in 74%, 21%, and 8% of CRC tissues. No significant correlation between three genes co-methylation and lymph node metastasis (2=0.725,P>0.05) was observed. The methylation of CDH1 gene, but not Syk or TIMP-3 gene, was significantly related with liver and lung metastasis of CRC (2=6.938,P<0.01). The methylation of Syk or TIMP-3, and co-methylation of CDH1 and TIMP-3 were risk factors in the prognosis of colorectal cancer after surgical treatment, and three genes co-methylation was not a risk factor. CONCLUSION: Our results indicate that methylation of multiple genes exists in CRC tissues. The methylation of multiple genes is not a necessary factor in the pathogenesis, metastasis and recurrence of CRC, and also it is not a necessary prognostic factor in CRC.     

Correlation between expression of ALDH1/ABCG2 and microvessel formation in epithelial ovarian cancer

AIM: To elucidate the correlation between the expression of aldehyde dehydrogenase 1 (ALDH1)/ATP-binding cassette subfaminly G member 2 (ABCG2) and microvessel density (MVD) in epithelial ovarian cancer (EOC).METHODS: In 198 specimens of EOC and 60 specimens of ovarian benign epithelial tumor tissues, the protein expression of ALDH1/ABCG2 and CD105 (microvessel marker) was detected by immunohistochemical staining.RESULTS: The positive rates of ALDH1 and ABCG2 in the EOC were 64.1% and 61.6%, respectively, while the positive rates in benign epithelial tumor tissues were 8.3% and 6.7%, respectively, and there were significant differences between them (P<0.05). In EOC and benign epithelial tumor tissues, the MVD were 22.6±9.7 and 5.03±3.35, respectively, and the difference was also significant (P<0.05). The expression of ALDH1 and ABCG2 in EOC was significantly related to differentiation, FIGO stage,and abdominal organ and lymph node metastasis (P<0.05). MVD had correlation with differentiation, FIGO stage, ascite, and abdominal organ and lymph node metastasis (P<0.05). MVD had positive correlation with the expression of ALDH1 and ABCG2 (P<0.01). There was also a positive correlation between the expression of ALDH1 and ABCG2 (P<0.01). Over-expression of ALDH1/ABCG2 and MVD≥23 were related to the poor prognosis. The survival rates in ALDH1/ABCG2 positive and MVD≥23 groups were significantly lower than those in ALDH1/ABCG2 negative and MVD<23 groups (P<0.05). The FIGO stage, the expression of ALDH1/ABCG2 and MVD were indepen-dent prognosis factors of EOC (P<0.05).CONCLUSION: The results suggest that the expression of ALDH1/ABCG2 and MVD in EOC are related to differentiation, lymph node metastasis, clinical stage and prognosis. Combined detection of these indexes may play an important role in predicting the progression and prognosis of EOC.     

Relationship between expression of COX-2 and clinicopathological features in esophageal carcinoma

AIM:To examine COX-2 expression in esophageal carcinoma, and to study relationships between COX-2 expression and clinicopathological features and prognosis of esophageal carcinoma patients. METHODS:89 paraffin - embedded tissue samples from patients with esophageal carcinoma were collected, its clinicopathological features such as tumour differentiation, depth of invasion, length and site of the tumor, regional lymph node metastases, distant metastasis were recorded. Survival time of 81 cases were also recorded. By SP immunohistochemistry method, the expression of COX-2 in tumor samples was examined. RESULTS:COX-2 expression in esophageal carcinoma was markedly higher than that in nomal esophagus, the expression was higher in less differentiated and deeper invaded cases (P<0.05), but it had no correlations with other clinicopathological features such as age,sex, length and site of the tumor, regional lymph node metastases, and distant metastasis (P>0.05). Cases of esophageal carcinoma with lower COX-2 expression had longer survival time than those with higher COX-2 expression (P<0.01). CONCLUSIONS:COX-2 expression is higher in esophageal carcinoma than normal esophagus. COX-2 expression of esophageal carcinoma is higher in less differentiated and deeper invaded cases, but it has no correlation with age, sex, length and site of the tumor, regional lymph node metastases, and distant metastasis. Patients with lower COX-2 expression have longer survival time than those with higher COX-2 expression.     

Abnormalities of miR-21 expression in human breast cancer is associated with distinctive pathologic features and shortened postoperative survival

AIM: To investigate the potential relevance of miR-21 expression level to clinicopathological characteristics and patient survival. METHODS: 113 BRCA cases with more then 5 years fallow-up data were selected. Total RNA from formalin-fixed paraffin-embedded (FFPE) tissues of 113 breast cancer (BRCA) and normal adjacent tissues (NATs) were isolated for miR-21 quantitative analysis by real-time RT-PCR. RESULTS: The miR-21 expression levels in BRCA were significantly higher than those in NATs (P<0.01) with average up-regulated level of 1.74 ± 0.48. Interestingly, high level expression of miR-21 was significantly correlated with advanced clinical stage (P<0.01), lymph node metastasis (P<0.01), and shorter survival of the patients ［hazard ratio (HR)=5.476, P<0.01］. Multivariate Cox regression analysis revealed that miR-21 was one of independent prognostic impacts (HR=4.133, P<0.01) on BRCA. CONCLUSION: Over-expression of miR-21 is associated with poor prognosis of BRCA and may serve as an independent prognostic marker for BRCA.