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.     

Expression and clinical significance of Bmi-1 in colorectal cancer

AIM:This study was to investigate the expression and significance of Bmi-1 in colorectal carcinoma (CRC), and to explore the effect of Bmi-1 on Ki67 expression in human CRC.METHODS:The samples from sixty CRC, thirty adenomas and twenty normal colorectal mucosal tissues were used in this study.The expression of Bmi-1 protein was detected by immunohistochemistry.The clinicopathological features and survival rate of patients were also analyzed.RESULTS:The overexpression of Bmi-1 was respectively 25.0％, 6.7％and 0% in CRC and adenomas as well as normal colorectal mucosal tissues.The results showed that the expression of Bim-1 was significantly higher in CRC, compared with that in adenomas and normal colorectal mucosal tissues (P<0.05).The overexpression of Bmi-1 protein in CRC was obviously associated with distant metastasis and TNM stage (P<0.05), but not with gender, age, tumor size, tumor site, histological type, differentiation degree and lymph node metastasis (P>0.05).Kaplan-Meier survival analysis showed that the overexpression of Bmi-1 reduced significantly survival of CRC patients (P<0.05).No statistical relation between expression of Bmi-1 and Ki67 in CRC was observed.CONCLUSION:The overexpression of Bmi-1 protein is significantly correlated with tumorigenesis, metastasis and prognosis of CRC.Bmi-1 might be regarded as a parameter in evaluating prognosis of CRC.     

Relationship between RUNX3,cyclin E,P21 and survival in gastric cancer patients

AIM:To evaluate the relationship between RUNX3,cyclin E,P21,biological features and survival in gastric cancer patients.METHODS:RUNX3 was examined using immunohistochemical staining.Cyclin E and P21 were analyzed by flow cytometry.Survival was evaluated by Kaplan-Meier survival curves.RESULTS:The positive-expression rate of RUNX3,cyclin E and P21 in tumor tissue from 56 patients with gastric cancer were 44.6%,64.3% and 32.1%,respectively.RUNX3 expression was correlated with lymph node metastasis and distant metastasis (P<0.05).Cyclin E might be related to depth of invasion,lymph node metastasis and distant metastasis (P<0.05).P21 was correlated with lymph node metastasis (P<0.05).It was revealed that RUNX3 and P21 were correlated (r=0.57,P<0.05),no correlation between RUNX3 and cyclin E was observed (r=0.25,P>0.05).Using Kaplan-Meier survival curves and the Log-rank test,there was correlation between RUNX3,cyclin E and survival (P<0.05).No correlation between P21 and survival was observed (P>0.05).CONCLUSION:RUNX3 may be related with tumorigenesis and tumor progression by affecting P21 expression.The detection of RUNX3 and cyclin E may be helpful in evaluating the clinicopathological parameters and prognosis in gastric carcinoma patients.     

Expression of bFGF in malignant tumor and its clinical pathological significance

AIM: To detect basic fibroblast growth factor (bFGF) expression in clinical common malignant tumor (non-small-cell lung cancer,breast cancer, colon cancer and melanoma), and to identify relationship between the expression and tumor clinicopathological characteristics.METHODS: Immunohistochemical SP method was used to detect the expression of bFGF at protein level in 208 cases of paraffin-embedded tissue of primary malignant tumor patients (68 cases of lung cancer, 80 cases of breast carcinoma, 41 cases of colon cancer and 19 cases of melanoma).RESULTS: The bFGF protein expression levels were significantly higher in low differentiated non-small-cell lung cancer with lymph node metastasis, and were positively correlated with TNM. In addition, no significant influence of the bFGF protein expression on the patients with median survival period was observed. The protein expression of bFGF was higher in advanced breast cancer with lymph node metastasis and was commonly found in the middle/higher differentiated colon cancer with regional lymph node metastasis. Meanwhile, bFGF protein was highly expressed in advanced melanoma patients with lymph node metastasis.CONCLUSION: bFGF may participate in the process of occurrence and progression of malignant tumor. Expression of bFGF protein may be an effective parameter for evaluating metastasis and prognosis of malignant tumor.     

Expression of Foxp3+ Tregs and PD1 in gastric cancer tissues and their correlation with clinicopathological factors and prognosis

AIM: To investigate the expression of Foxp3⁺ regulatory T cells (Foxp3⁺ Tregs) and programmed death receptor 1 (PD1) in gastric cancer tissues and their association with clinicopathological factors and prognosis of the patients. The correlation between the 2 molecules was also analyzed at the same time. METHODS: The tumor sections from 111 gastric cancer patients were stained for Foxp3 and PD1 by the method of immunohistochemistry. The associations of the expression levels of these 2 molecules with clinicopathological factors involved in the disease progression and prognosis were statistically analyzed. The relationship of their expression was detected. RESULTS: Foxp3⁺ Tregs and PD1 were expressed in the gastric cancer tissues, and PD1 was expressed in the tumor infiltrating lymphocytes (TILs). The expression of Foxp3 and PD1 was correlated with lymph node metastasis, clinicopathological stage and prognosis of gastric cancer patients. The expression of these 2 determinants in the patients with lymph node metastasis and an advanced clinicopathological stage was distinctly higher (P <0.05). The patients with positive expression of the 2 indexes presented a lower overall survival rate and worse prognosis (P <0.05). A significantly positive correlation between the infiltration of Foxp3⁺ Tregs and the expression of PD1⁺ TILs was also observed (P <0.01).CONCLUSION: Foxp3⁺ Tregs and PD1⁺ TILs co-infiltrate in the gastric cancer tissues, which can be used as biological markers to predict the disease progression and prognosis.     

Expression of miR-143 and its clinicopathologic significance in gastric cancer

AIM: To investigate the expression of miR-143 and its association with clinicopathologic features in gastric cancer.METHODS: The expression level of miR-143 in 32 cases of gastric cancer and matched non-tumor adjacent tissue specimens was examined using stem-loop real-time RT-PCR. The relationship between the expression of miR-143 and its clinicopathologic features of gastric cancer was analyzed.RESULTS: The expression level of miR-143 was significantly lower in the tumor tissues than that in the adjacent tissues (P<0.05). Down-regulated miR-143 expression was associated with the cell differentiation (P<0.05) and lymph node metastasis (P<0.05) in gastric cancer patients. No significant association was found between the expression of miR-143 and the status of gender, age, blood type, tumor location, tumor size, depth of tumor invasion and tumor node metastasis stage.CONCLUSION: It is possible that miR-143 plays an important role in the generation and progression of gastric cancer. The expression level of miR-143 may be a valuable adjuvant parameter for predicting the poorly differentiated gastric cancer.     

Study of annexin A2 expression in gastric cancer by proteomics and tissue microarray

AIM: To investigate the differential expression of annexin A2 (ANXA2) in gastric carcinoma and to analyze the relationship between ANXA2 expression and clinicopathological parameters of gastric carcinoma. METHODS: Pure gastric adenocarcinoma cells (GAC) and normal gastric epithelial cells (NGEC) in 15 patients with gastric cancer were acquired by laser capture microdissection (LCM). All peptide specimens after trypsin digestion were labeled with ¹⁸O/¹⁶O. Quantitatively identification of differential expression of the proteins betweem GAC and NGEC was performed by Nano-RPLC-MS/MS. The expression of ANXA2 in the 2 kinds of tissues was detected by Western blotting. Tissue microarray containing 75 pairs of gastric carcinoma and para-carcinoma tissues was used and the expression of ANXA2 in these specimens was detected by the method of immunohistochemistry (IHC). The relationship between ANXA2 expression and clinicopathological parameters of the pateints with gastric carcinoma was analyzed. RESULTS: A total of 78 differential proteins were identified and ANXA2 was up-expressed in GAC (2.32∶ 1), which was confirmed by Western blotting (P<0.01). The results of IHC showed that the correlations between the expression level of ANXA2 protein and invasive depth (T stage), lymph node metastasis (N stage), histological differentiation, TNM stage and the size of tumor were observed (P<0.01), but the correlations between the ANXA2 expression and sex, age and distant metastasis (M stage) were not found (P>0.05). CONCLUSION: The up-expressed ANXA2 may play an important role in the biological behavior of gastric cancer.     

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.     

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 genetic instability of nm23H1 gene and clinical pathological behaviors in gastric cancer and colonic cancer

AIM: To study the relationship between genetic instability of nm23H1 gene and clinical pathological behaviors in Chinese with gastric cancer and colonic cancer, and provide experimental basis for the mechanism of nm23H1 gene and tumor metastasis. METHODS: This study was conducted on 40 gastric carcinomas and 30 colonic carcinomas. Techniques such as DNA extraction from formalin-fixed paraffin-embedded tissues, PCR-SSCP, ordinary silver stain were used to study microsatellite instability (MSI) and loss of heterozygosity (LOH) of locus D17S396. Envision immunohistochemistry and Leica-Qwin computer imaging techniques were used to assess the expression of nm23H1 protein. RESULTS: In both gastric cancer and colonic cancer, the frequency of MSI was higher in TNM stageⅠandⅡthan that in stage Ⅲ and Ⅳ, while LOH was just opposite. Moreover, the frequency of LOH in lymph node metastasis cases was significantly higher than that without lymph node metastasis cases. The positive frequency of nm23H1 protein with lymph node metastasis was lower than that without lymph node metastasis cases. TNM stage III and IV also exhibited lower nm23H1 protein positive frequency compared with stage I and II. CONCLUSION: MSI and LOH can control the carcinogenesis and metastasis of gastric cancer and colonic cancer through different approaches. MSI may be an early period molecule marker of gastric cancer and colonic cancer. In contrast, LOH appears mostly in the late period of gastric cancer and colon cancer, indicating a high aggressive and poor prognosis.     

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.     

Expression of CD44s in lung cancer and its significance

AIM: To investigate expression of CD44s in lung cancer and it's clinical significance. METHODS: A total of 117 primary lung cancer from patients were examined for CD44s expression by immunohistochemical staining. RESULTS: CD44s mostly expressed in non-small cell lung cancer (NSCLC) but not in small ecll lung cancer (SCLC), and squamous cell carcinoma(SCC) showed much stronger expression of CD44s than adenocarcinoma(ADC)(P＜0.05). In comparison of the lung cancer with/ without lymph node metastasis, the latter showed stronger expression of CD44s(P＜0.01). According to TNM, there was a distinct statistic difference between early stage and advanced stage(P＜0.05). CONCLUSION: CD44s might be a better indicant in histological classification of lung cancer, lymph node metastasis, clinical stage and prognosis.     

Central role of GRP78 in growth of gastric carcinoma cells

AIM: To explore the effect of glucose-regulated protein 78 (GRP78) on the gastric carcinogenesis. METHODS: GPR78 expression patterns were examined in 34 specimens from gastric carcinoma patients using the immunohistochemistry (IHC) assay, and in 10 specimens using Western blotting analysis. In addition, the expression of GPR78 and cyclin D1 was detected in human gastric cancer cell lines SGC7901 and SGC7901-H78 (overexpressing GRP78) by Western blotting. RESULTS: By IHC assay, GRP78 was found to be highly expressed in the cytoplasm of gastric carcinomas as compared with the adjacent non-malignant tissues and corresponding normal tissues. GRP78 expression was positively correlated with gender and histological differentiation (P<0.05), but not with age, tumor stage and lymph node metastasis (P>0.05). Furthermore, we found that with the increased expression of GRP78 in SGC7901-H78 cells, the expression of cyclin D1 was also elevated. CONCLUSION: GRP78 might be a key player to be involved in the growth of gastric cancer.     

Expression of chemokine receptor CCR7 and VEGF-C is associated with prognosis of breast carcinoma

AIM: To explore the protein levels of chemokine receptor 7 (CCR7) and vascular endothelial growth factor (VEGF)-C in breast carcinoma, and to investigate the effects of CCR7 and VEGF-C on prognosis of breast carcinoma. METHODS: The protein expression levels of CCR7 and VEGF-C in the breast carcinoma tissues and normal breast tissues were detected by the method of immunohistochemistry. At the same time, the relationship between clinicopathologic characteristics and the protein expression of CCR7 and VEGF-C in the breast carcinoma tissues was analyzed. The relationship between the protein expression of CCR7 and VEGF-C and survival time of the breast cancer patients was estimated by Kaplan-Meier method.RESULTS: The positive expression rates of CCR7 and VEGF-C in the breast carcinoma tissues were significantly higher than those in the normal breast tissues (P<0.01). A positive correlation was observed between the protein expression of CCR7 and the protein expression of VEGF-C in the breast carcinoma tissues (r=0.613, P<0.01). The protein expression of CCR7 and VEGF-C was correlated with lymph node metastasis and TNM stage (P<0.05), but both were not related to patients' age, primary tumor size, estrogen receptor and progesterone receptor. The survival time of the patients with CCR7 and VEGF-C positive expression was significantly shorter than that of the patients without the expression (P<0.05).CONCLUSION: The positive expression of CCR7 and VEGF-C proteins is associated with the prognosis of breast cancer, and combined detection of CCR7 and VEGF-C protein expression levels may be helpful to judge the prognosis of breast cancer.     

Analyzing clinical characteristic of miR-196b in human colorectal cancer using TCGA and effect of miR-196b on 5-FU resistance of CRC cell line

AIM: To investigate the clinical characteristics of microRNA (miR)-196b in colorectal cancer (CRC) and to study its biological function in 5-fluorouracil (5-FU) resistance. METHODS: miRNA sequence dataset and the corresponding clinical data of CRC patients were downloaded from The Cancer Genome Atlas (TCGA). Expression level and clinical characteristics of miR-196b in CRC patients were analyzed using SPSS 17.0. CRC cell line overexpres-sing miR-196b was established using transient transfection method. MTS test was used to evaluate the effect of miR-196b overexpression on 5-FU resistance. RESULTS: miR-196b expression was associated with lymph node metastasis and TNM stage (P<0.05), but not related with age and sex. Lymph node metastasis and distant metastasis were independent prognostic factors for rectal patients (P<0.05). The expression level of miR-196b was not associated with survival condition of rectal patients. The viability of the cells overexpressing miR-196b treated with different concentrations of 5-FU was significantly higher than that in control group (P<0.05). CONCLUSION: miR-196b may be a potential biomarker of TNM stage and lymph node metastasis in CRC. miR-196b increases the 5-FU resistance of CRC cells.     

Quantitative perfusion parameters of dynamic contrast-enhanced magne-tic resonance imaging in patients with rectal cancer: estimation of the microvascular perfusion and permeability

AIM: To investigate the perfusion parameters using dynamic contrast-enhanced magnetic resonance imaging (DCE-MRI) in rectal cancer patients so as to explore its potential value in estimating the microvascular condition including perfusion and permeability. METHODS: The data of 38 rectal cancer patients examined with DCE-MRI was retrospectively analyzed. The perfusion parameters of carcinoma and normal rectal wall in each case were calculated, including volume transfer constant (K^trans), rate constant of back flux (K_ep), extravascular extracellular space fractional volume (V_e) and initial area under curve (iAUC). The mean values of tumor and normal rectal wall, mucinous and nonmucinous carcinoma, poorly and moderately-to-well differentiated carcinoma, case with or without lymph node metastasis were compared. RESULTS: All the parameters of rectal cancer were higher than normal rectal wall (P<0.01). No significant difference was found between poorly and moderately-to-well differentiated carcinoma in terms of K^trans, K_ep and V_e, neither was the case with or without lymph node metastasis. The cases with lymph node metastasis had lower iAUC than those without (P<0.05). CONCLUSION: Quantitative perfusion DCE-MRI answered the microvascular perfusion and permeability change of rectal cancer compared with normal rectal wall, besides it could be used to distinguish between mucinous and nonmucinous carcinoma, which demonstrated its value in the evaluation of rectal cancer. However, it should not be recommended to predict the degrees of tumor cell differentiation and lymph node metastasis just according to the perfusion parameters.