Expression In Colorectal Carcinoma Research Articles

Prognostic significance of c-erbB-2 and p53 expression in colorectal carcinoma

Objective: Colorectal cancer is the fourth commonest form of cancer occurring worldwide. In order to recognize the prognosis in colorectal carcinoma, a lot of clinic and pathologic parameters such as; tumor size, microscopic type, microscopic grade, depth of invasion, lymph node involvement, invasion of vascular and perineural, tumor stage have been used. In this study, we aimed to compare expression of c-erbB-2, p53 and classical prognostic factors, and to investigate the correlation with prognosis. Methods: In our study, operation materials belonging to 50 colorectal cases were examined retrospectively. Results: Overall, 100 % of colorectal carcinomas had nuclear-staining for p53 protein and 78% of carcinomas cytoplasmic and membranous staining or the c-erbB-2 showing distribution in a heterogeneous fashion. p53 overexpression correlated with (p<0.005) pathologic stage, metastasis of lymph node, invasion of angiolymphatic, differentiation and degree of tumors. c-erbB-2 overexpressions correlated with (p<0,005) histologic differentiation and peritumoral lymphocytic infiltration but not correlated with tumor size, tumor growth pattern, localisation of tumors, pathologic stage, metastasis of lymph node, invasion of angiolymphatic. Conclusion: These results suggest that evaluation of c-erbB-2 and p53 Immunohistochemical expression in colorectal carcinomas may be of value in predicting the clinical course in these patients.

Immunohistochemistry expression of TCF4 protein on carcinoma, adenoma and non neoplastic colorectal mucosa

PurposeTo detect and quantify the immunoreactivity of TCF4 protein in colorectal carci- noma, colorectal adenoma and non-neoplasic colorectal epithelium. MethodsWe studied 129 individuals: 40 with colorectal cancer, 52 with colorectal ad- enoma and 37 with non-neoplastic colorectal epithelium. The colorectal adenoma and carcinoma samples were obtained from patients who underwent surgical procedures, and colonoscopies and samples of non-neoplastic colorectal epithelium were taken from patients who died from cardiovascular diseases, without diseases of the large intestine. Samples of different tissues were included in paraffin blocks, and the immunohistochem- ical expression of protein TCF4 was analyzed using the technique of tissue microarray (TMA) with polyclonal antibody TCF4. The immunoreactivity was analyzed and classified as positive and negative. ResultsThe immunohistochemical expression of TCF4 protein was significantly higher (p < 0.01) in colorectal carcinoma than in the non-neoplastic colorectal epithelium and adenoma. There was no difference (p = 0.76) between TCF4 protein immunohistochemical expression in colorectal adenoma and non-neoplastic colorectal tissue. ConclusionsTCF4 protein showed a more intense expression in colorectal carcinoma than in non-neoplastic colorectal epithelium and adenoma, indicating that this protein is in- volved in colorectal carcinogenesis.

HLA Class II Antigen Expression in Colorectal Carcinoma Tumors as a Favorable Prognostic Marker

The goal of this study was to determine the frequency of HLA class II antigen expression in colorectal carcinoma (CRC) tumors, its association with the clinical course of the disease, and the underlying mechanism(s). Two tissue microarrays constructed with 220 and 778 CRC tumors were stained with HLA-DR, DQ, and DP antigen-specific monoclonal antibody LGII-612.14, using the immunoperoxidase staining technique. The immunohistochemical staining results were correlated with the clinical course of the disease. The functional role of HLA class II antigens expressed on CRC cells was analyzed by investigating their in vitro interactions with immune cells. HLA class II antigens were expressed in about 25% of the 220 and 21% of the 778 tumors analyzed with an overall frequency of 23%. HLA class II antigens were detected in 19% of colorectal adenomas. Importantly, the percentage of stained cells and the staining intensity were significantly lower than those detected in CRC tumors. However, HLA class II antigen staining was weakly detected only in 5.4% of 37 normal mucosa tissues. HLA class II antigen expression was associated with a favorable clinical course of the disease. In vitro stimulation with interferon gamma (IFNγ) induced HLA class II antigen expression on two of the four CRC cell lines tested. HLA class II antigen expression on CRC cells triggered interleukin-1α (IL-1α) production by resting monocytes. HLA class II antigen expression in CRC tumors is a favorable prognostic marker. This association may reflect stimulation of IL-1α production by monocytes.

An evaluation of nuclear factor kappa B expression in colorectal carcinoma: an analysis of 50 cases.

There is a rising trend in the incidence of (colorectal carcinoma) colorectal cancer (CRC) in India. Nuclear factor kappa B (NFkB) is a transcription factor which belongs to the Rel family. It has an impact on phenomena such as apoptosis, tumor progression and differentiation. (1) To evaluate the grade and stage in 50 cases of colorectal carcinoma. (2) To evaluate the NFkB translocation into the nucleus of the cells. (3) To compare the benign and malignant areas with the degree of NFkB translocation and compare the translocation with the grade and pathological stage. The grade and stage of the tumors was evaluated. NFkB staining was performed on the tissues. The results of the immunostaining were analyzed semi quantitatively as a percentage of positive cells. Statistical Package for Social Sciences 13.0 statistical package program (SPSS, Lead Technologies Inc, USA) for windows was used. Correlation was carried out using the Pearson's correlation co-efficient and the Chi-square test. There were 29 males (58%) and 21 females (42%). Most of the cases were well-differentiated adenocarcinomas (58%). There was a significant difference between NFkB translocation in the epithelial cells and lymphocytes in the benign and malignant areas (P - 0.04 and P - 0.001 respectively). There was a significant correlation between the grade of NFkB staining in the malignant epithelial cells with the tumor and nodal status (P - 0.001 and P - 0.001). It is likely that NFkB is an important factor in the pathogenesis of CRC. Further studies including therapeutic intervention using strategies which prevent activation of NFkB in colorectal carcinoma patients will tell if we could alter the course of the disease favorably.

Expression and clinical significance of neuropilin-2 in colorectal cancer: correlation with lymphangiogenic factors

Purpose: Neuropilin-2 is expressed by venous and lymphatic endothelial cells and can bind the lymphangiogenesis-associated ligand vascular endothelial growth factor (VEGF)-C and D. Recently, neuropilin-2 was shown to play a role in lymph node metastasis and cancer promotion. The aim of this study was to determine the clinical relevance of neuropilin-2 expression in colorectal carcinomas. Methods: Immunohistochemical analysis of neuropilin-2 expression of 350 colorectal cancer samples was evaluated using tissue microarray. The relationship between neuropilin-2 expression and other clinicopathological variables were analyzed in colorectal carcinomas. To clarify the role of neuropilin-2 in colorectal cancer lymphangiogenesis, its expression was examined with VEGF-C, VEGF-D, and VEGF receptor 3 expression of the colorectal carcinomas. Results: Neuropilin-2 was detected in 176 of 350 colorectal carcinomas (50.3%). Overexpression of neuropilin-2 was correlated with lymph node metastasis (P=0.041) and VEGF-D expression (P=0.020), but was not correlated with TNM (tumor, node, metastasis) stage, other lymphangiogenic factor expression and survival colorectal cancer patients. Conclusion: Neuropilin-2 expression was correlated with lymph node metastasis and VEGF-D expression in colorectal carcinomas. Our results suggest a possible role of neuropilin-2 in lymphangiogenesis and cancer progression in colorectal carcinomas.

Trefoil factor 1 gene alternations and expression in colorectal carcinomas

Aberrant expression of the trefoil factor family (TFF) has been recognized to be involved in the development and/or progression of various solid tumors. Increased trefoil factor 1 (TFF1) expression is found associated with tumor progression in some tumors, and TFF1 missense mutations have been detected in gastric cancer. The aim of the study was to analyze TFF1 alternations and expression in colorectal carcinoma and their correlation with cancer progression and pathological aspects. TFF1 mutations were detected in colorectal carcinomas by DNA sequencing. TFF1 mRNA and protein levels in subsets of the primary tumors were determined using quantitative reverse transcription polymerase chain reaction and immunohistochemistry analyses. The serum level of TFF1 was also detected by enzyme-linked immunosorbent assay for patients with colorectal carcinoma. Five variants were detected in the 5'-untranslation region and intron 1 of TFF1. TFF1 expression was increased in colorectal carcinoma compared to paired distal colonic mucosa. Immunohistochemistry in primary colorectal carcinoma showed no significant differences in tumor TFF1 levels with respect to clinicopathological parameters such as the patient's sex, cancer differentiation, stage and lymph node metastasis. However, serum TFF1 levels were significantly elevated in patients with colorectal carcinoma compared to healthy individuals. The results indicate that TFF1 missense mutations seem to be a rare event in colorectal carcinogenesis. Serum TFF1 may be a potential useful marker for patients with colorectal carcinoma.

Clinical impacts of mammalian target of rapamycin expression in human colorectal cancers

This study investigated the clinicopathologic roles of mammalian target of rapamycin (mTOR) expression and its relationship to carcinogenesis and tumor progression in a colorectal adenoma-adenocarcinoma model. Two colon cancer cell lines with different pathologic stages (SW480 and SW48) and 1 normal colonic epithelial cell line (FHC) were used, in addition to 119 colorectal adenocarcinomas and 32 adenomas. mTOR expression profiles at messenger RNA (mRNA) and protein levels were investigated in the cells and tissues using real-time quantification polymerase chain reaction and immunohistochemistry. The findings were correlated with the clinicopathologic features of the tumors. The colon cell line from stage III cancer (SW48) showed higher expression of mTOR mRNA than that from stage II cancer (SW480). At the tissue level, mTOR showed higher mRNA and protein expression in colorectal carcinoma than in adenoma. The mRNA and protein expression was correlated with each other in approximately one-third of the carcinomas and adenomas. High levels of mTOR mRNA expression were noted more in carcinoma or adenoma arising from the distal portion of the large intestine (P = .025 and .019, respectively). Within the colorectal cancer population, a high level of expression of mTOR mRNA was related to the presence of lymph node metastases (P = .031), advanced pathologic stage (P = .05), and presence of persistent disease or tumor recurrence (P = .035). To conclude, the study has indicated that mTOR is likely to be involved in the development and progression of colorectal cancer and is linked to cancer initiation, invasiveness, and progression.

Physical Activity, Tumor PTGS2 Expression, and Survival in Patients with Colorectal Cancer

Higher levels of physical activity are associated with lower colorectal carcinoma incidence and mortality, perhaps through influencing energy balance, cellular prosta7 systemic inflammation. Although evidence suggests interactive effects of energetics, sedentary lifestyle, and tumor CTNNB1 (β-catenin) or CDKN1B (p27) status on colon cancer prognosis, interactive effects of physical activity and tumor PTGS2 (the official symbol for COX-2) status on clinical outcome remain unknown. Using molecular pathological epidemiology database of 605 stage I-III colon and rectal cancers in two prospective cohort studies (the Nurse's Health Study and the Health Professionals Follow-up Study), we examined patient survival according to postdiagnosis physical activity and tumor PTGS2 status (with 382 PTGS2-positive and 223 PTGS2-negative tumors by immunohistochemistry). Cox proportional hazards models were used to calculate colorectal cancer-specific mortality HR, adjusting for clinical and other tumor variables including microsatellite instability status. Among PTGS2-positive cases, compared with the least active first quartile, the multivariate HRs (95% confidence interval) were 0.30 (0.14-0.62) for the second, 0.38 (0.20-0.71) for the third, and 0.18 (0.08-0.41) for the fourth quartile of physical activity level (Ptrend = 0.0002). In contrast, among PTGS2-negative cases, physical activity level was not significantly associated with survival (Ptrend = 0.84; Pinteraction = 0.024, between physical activity and tumor PTGS2 status). Postdiagnosis physical activity is associated with better survival among patients with PTGS2-positive tumors but not among patients with PTGS2-negative tumors. Immunohistochemical PTGS2 expression in colorectal carcinoma may serve as a predictive biomarker in pathology practice, which may predict stronger benefit from exercise.

Research development of the relationship between thymidine phosphorylase expression and colorectal carcinoma.

Thymidine phosphorylase (TP) is a key enzyme that contributes to the composition and decomposition of pyrimidine nucleotides. TP seems homologous to platelet-derived endothelial cell growth factor, and its effects on inducing vascularization and anti-apoptosis are closely related to growth and metastasis of colorectal carcinoma. In addition, TP is a key enzyme that catalyzes the transformation from 5-fluorouracil (FU) prodrugs of 5′-deoxy-5-fluorouridine (5′-DFUR) to 5-FU. The activity of TP is closely related to the sensitivity of colorectal carcinoma cells to fluorouracil drugs and targeted therapy. Given the important functions of TP in growth, metastasis, tumor treatment, and prognosis, determining its expression mechanism is significant. This article summarizes the research development of TP expression in colorectal carcinoma, tumor neovascularization, cytotoxicity activation of 5′-DFUR, and colorectal carcinoma therapy.

Significance of Secreted Protein Acidic and Rich in Cysteine Expression in Colorectal Carcinoma

See Article on Page 93-99 The five hallmark capabilities of cancer cell for invasion and metastasis are self sufficiency in growth signal, insensitivity to growth inhibitory signals, evasion of apoptosis, limitless replicative potential and sustained angiogenesis [1]. There were several consistently appeared metastsasis risk factors of primary tumor with poor prognosis such as poor histologic grade, depth of invasion, and lymphovascular invasion. The stromal compartment is a complex arrangement of stromal cells and extracellular matrix, plus associated growth factors, regulatory molecules, and remodeling enzymes. Blood vessels, nerves, and immune cells are also integral parts of the stroma. These stromal components regulate cell function and maintain overall tissue homeostasis [2]. In cancer, the tumor microenvironment directly influences the growth of the tumor and its ability to progress and metastasis. Several proteins such as E-cadherin, matrix metalloproteinase were changed during invasion and metastasis. Down regulation of E-cadherin stimulated by many growth factors such as epidermal growth factor receptor, c-MET, fibroblast growth factor receptor, Src-family kinase, insulinlike growth factor-I receptor in tumor cells promote tumor invasion and metastasis. SPARC is one of the matricellular proteins and has the important role in interaction between the cells and matrix. The apparent contradictory functions of SPARC were reported in angiogenesis, epithelial-to-mesenchymal transition and invasion through matrix metalloproteinase (MMP) according to different sites of cancers. Expression of SPARC was significantly correlated with the expression of vascular endothelial growth factor (VEGF) and microvessel density in colon cancer tissues [3]. In ovarian cancer, the absence of SPARC upregulated the expressions of VEGF, VEGFR2, MMP-2, and MMP-9, thereby promoting the angiogenic and metastatic potential of these cancers [4]. SPARC is also able to inhibit VEGF-induced integrin activation and down regulation of MMP-2 and MMP-9. There were several reports on the role of SPARC as a prognostic factor of colon cancer patients. A survival analysis of colorectal carcinoma patients revealed a poorer prognosis for patients lacking SPARC expression than for patients with normal SPARC expression [5]. Patients with low or absence expressing SPARC had significantly worse overall survival and disease-free survival. The role of SPARC in colorectal cancer cell and stroma as prognostic makers need further study after evaluation of monoclonal antibodies of SPARC because of different results [6]. SPARC has been shown to modulate response to chemotherapy. Addition of exogenous SPARC and/or forced expression of endogenous SPARC improved colon cancer cell response to chemotherapy in vitro [7]. Lower levels of SPARC expression in colorectal cancers may represent a therapy-resistant phenotype, and that biologically, low levels of SPARC negatively impacts a tumor's response to chemotherapy [8]. In the future, manipulating host-tumor interactions with the matricellular proteins such as SPARC may be important in preventing carcinogenesis or metastasis, and in treating carcinoma by re-establishing normal control mechanisms [9] or reducing drug resistance.

Prognostic relevance of BRD7 expression in colorectal carcinoma

BRD7 is a member of bromodomain-containing protein and was found to be a cofactor of P53. Down-regulation of BRD7 has been shown in nasopharyngeal carcinoma cell lines and tissues. However, the clinical role of BRD7 in colorectal cancer remains unknown. Real-time PCR, Western blotting analysis and immunohistochemistry were employed to examine BRD7 expression in CRC cell lines/tissues compared with normal epithelia cells/adjacent non-tumorous tissues. In addition, statistical analyses were applied to evaluate the diagnostic value and associations of BRD7 expression with clinical parameters of patient samples. BRD7 was down-regulated in colorectal cancer cell lines and cancerous tissues compared with that in normal colon epithelial cells and adjacent noncancerous tissue samples. BRD7 protein expression was positively correlated with clinical stage (P < 0·001), T classification (P = 0·001), N classification (P < 0·001), M classification (P< 0·001) and pathologic differentiation (P= 0·008). Patients with low/none BRD7 expression had shorter overall survival time than those with higher BRD7 expression. Univariate and multivariate analyses indicated BRD7 expression was an independent prognostic factor (P<0·001). BRD7 may serve as a potential prognostic biomarker of human colorectal cancer.

Expression of Cyclin D1 Is Associated with β-Catenin Expression and Correlates with Good Prognosis in Colorectal Adenocarcinoma

The aim of this study is to investigate the prevalence and prognostic impact of β-catenin and cyclin D1 expression in colorectal carcinoma (CRC) patients. We evaluated immunohistochemial expression of β-catenin and cyclin D1 using 2-mm cores from 220 CRC patients for tissue microarray, and its significance was statistically evaluated. Positive expression of β-catenin and cyclin D1 was found in 72.5% (158 of 218 cases) and 59.4% (129 of 217 cases) of CRC patients, respectively. Expression of β-catenin was significantly correlated with tumor location (P = .017), differentiation (P = .010), lymph node metastasis (P = .032), preoperative carcinoembryonic antigen level (P = .032), and cyclin D1 expression (P = .005). Expression of cyclin D1 was significantly correlated with recurrence and/or metastasis (P = .004). In univariate analysis, β-catenin expression predicted more favorable overall survival (P = .022) and cyclin D1 expression predicted both more favorable overall survival and relapse-free survival (P = .004 and P = .006, respectively). Multivariate analysis showed that tumor stage and expression of cyclin D1 were independent prognostic factors significantly associated with overall survival and relapse-free survival. This study shows that expression of β-catenin and cyclin D1 is associated with favorable clinicopathologic variables and it is a clinically significant prognostic indicator for CRC patients.

Usefulness of epithelial cell adhesion molecule expression in the algorithmic approach to Lynch syndrome identification

Lynch syndrome (LS), the most frequent form of hereditary colorectal cancer syndrome, is caused by germ-line mutations in the mismatch repair system genes. Recently, a new mechanism involving the epithelial cell adhesion molecule (EPCAM)/TACSTD1 gene has been shown to be responsible in cases with abnormal MSH2 expression. Of interest, 3' exons deletions of the EPCAM gene, which is located upstream of MSH2 in chromosome 2, are associated with MSH2 promoter hypermethylation. EPCAM protein, expressed in epithelial tissues, is encoded by the EPCAM/TACSTD1 gene. Our study's aim was to explore EPCAM expression in colorectal carcinomas of MSH2-associated LS cases to evaluate the usefulness of EPCAM protein expression in the algorithm approach to LS population screening. We included a total of 19 MSH2-negative colorectal carcinomas from 14 different patients in whom we were able to perform a complete germ-line analysis. Nine patients showed a deleterious germ-line mutation that involved the MSH2 gene in 3 instances and the EPCAM gene exon 9 in 6 instances. All patients harboring the EPCAM mutation belonged to the same family. Of the 19 colorectal carcinomas, EPCAM expression loss was seen in only 5 tumors, all of them from patients showing a germ-line EPCAM deletion. Of interest, 6 tumors from 3 different patients carrying the same germ-line EPCAM deletion showed normal EPCAM expression. In conclusion, owing to the high specificity of EPCAM protein expression to identify LS patients carrying an EPCAM deletion, we recommend adding EPCAM immunohistochemistry to the LS diagnostic algorithm in MSH2-negative colorectal carcinoma.

C-Kit and Musashi-1 expression in colorectal carcinoma and its association with etiological factors

c-Kit and Musashi-1 expression in colorectal carcinoma and its association with etiological factors

Clinicopathological and immunohistochemical study of mismatch repair gene (hMLH1 and hMSH2) expression in colorectal carcinoma of young Egyptian patients

Clinicopathological and immunohistochemical study of mismatch repair gene (hMLH1 and hMSH2) expression in colorectal carcinoma of young Egyptian patients

The clinicopathological significance of REIC expression in colorectal carcinomas.

REIC is down-regulated in immortalized cell lines compared with the parental normal counterparts, and could inhibit colony formation, tumor growth and induce apoptosis. Here, its expression was examined by immunohistochemistry on tissue microarray containing colorectal non-neoplastic mucosa (NNM), adenoma and adenocarcinoma. Colorectal carcinoma tissue and cell lines were studied for REIC expression or its secretory level by Western blot, RT-PCR or enzyme-linked immunosorbent assay (ELISA). The results demonstrated that REIC was differentially expressed in Colo201, Colo205, DLD-1, HCT-15, HCT-116, HT-29, KM-12, SW480, SW620, and WiDr with its secretion concentration less than 300 pg/mL. Carcinomas showed statistically lower REIC expression than matched NNM with no difference for protein content. Immunohistochemically, REIC expression was significantly decreased from NNM, adenoma to adenocarcinoma (p<0.05). REIC expression was negatively correlated with depth of invasion, TNM staging, dedifferentiation, Capase-3 and nuclear inhibitor of growth 5 (ING5) expression (p<0.05), while not with age, sex, tumor size, lymphatic or venous invasion, or lymph node metastasis (p>0.05). Kaplan-Meier analysis indicated that REIC expression was not associated with the prognosis of colorectal carcinomas (p>0.05). Cox's analysis demonstrated that lymphatic and venous invasion, lymph node metastasis, and UICC staging were independent prognostic factors for carcinoma (p<0.05). Our study indicated that down-regulated REIC expression might play an important role in colorectal adenoma-adenocarcinoma sequence and subsequent progression. Aberrant REIC expression might be employed as a good marker of pathogenesis and development of colorectal carcinomas.

IL-23 directly enhances the proliferative and invasive activities of colorectal carcinoma

Interleukin-23 (IL-23) plays an essential role in the mucosal immune system. It has been suggested that IL-23 is able to induce carcinogenesis as well as inflammation and a recent study revealed that IL-23R is expressed in colorectal carcinoma cells. However, neither the differences in the IL-23R expression among the patients nor the concrete functions of IL-23 in colorectal carcinoma cells have been revealed. The aim of the present study was to examine the characteristics of IL-23R expression in colorectal carcinoma and the direct effects of IL-23 on colorectal cancer cells. We examined the IL-23R expression in human colorectal cancer tissue samples by immunohistochemistry. Cell proliferation and invasion assays under IL-23 stimulation were performed using cultured cells derived from colorectal cancer. ELISA and real-time PCR were used to evaluate the transforming growth factor (TGF)-β production due to IL-23 stimulation. All of the TNM stage IV patients were positive for IL-23R. IL-23R expression in the carcinoma tissue was also relatively high at the deepest point of invasion in certain cases. The proliferative and invasive activities and/or TGF-β production of DLD-1 cells increased by IL-23 stimulation, whereas no change was observed in the activities of MIP101 and KM12c cells. IL-23 directly enhanced the malignancy of the colon carcinoma cells. An autocrine mechanism via TGF-β production may underlie these effects. IL-23 is therefore a potential target for cancer immunotherapy. However, the homogeneity in IL-23R expression and the effects of IL-23 on colorectal carcinoma cells should be considered.

A novel sialyl LeX expression score as a potential prognostic tool in colorectal cancer

BackgroundTreatment decisions in colorectal cancer subsequent to surgery are based mainly on the TNM system. There is a need to establish novel prognostic markers based on the molecular characterization of tumor cells. Evidence exists that sialyl LeX expression is correlated with an unfavorable outcome in colorectal cancer. The aim of this study was to establish a simple sialyl LeX staining score and to determine a potential correlation with the prognosis in a series of advanced colorectal carcinoma patients.MethodsIn order to implement routine use of sialyl LeX immunohistology, we established a new, easily reproducible score and defined a cutoff which discriminated groups with better or worse outcome, respectively. We then correlated sialyl LeX expression of 215 UICC stage III and IV patients with disease-free and cancer-related survival.ResultsA five-stage score could be established based on automated immunohistochemical stainings. Using a statistical model, we calculated a cutoff to discriminate between weak and strong staining positivity of sialyl LeX. Patients with strong positive specimens had a worse cancer-related survival (P = 0.004) but no difference was observed for disease-free survival (P = 0.352).ConclusionsThese results demonstrate a strong correlation between high sialyl LeX-expression in colorectal carcinomas and cancer-related survival. Our highly standardized and easy-to-use staining score is suitable for routine use and hence it could be recommended to evaluate sialyl LeX-expression as part of the standard histopathological analysis of colorectal carcinomas and to validate the score prospectively based on a larger population.

Downregulation of Prominin 1/CD133 expression in colorectal cancer by NSAIDs following short-term preoperative treatment

Expression of Prominin 1/CD133 is associated with poor prognosis and chemoresistance in several types of solid tumors. The aim of the present study was, therefore, to evaluate Prominin 1/CD133 expression in colorectal carcinoma after short-term preoperative treatment by non-steroidal anti-inflammatory drugs (NSAIDs). Patients aimed at curative operation for colon cancer were randomized to receive NSAIDs (indomethacin 50 mg x2 or celecoxib 100 mg x2) three days preoperatively. Antisecretory prophylaxis (esomeprasol 40 mg x1) was provided to all patients and served as sham intake. CD133 expression in tumor tissue was also assessed in tumors from Dukes' B patients selected for either long or short postoperative survival. No patients received perioperative chemoradiotherapy. Tumor tissue was collected at surgery for quantification of mRNAs (Prom1 and Wnt4) by qPCR. Immunohistochemistry stained for CD133, Ep-CAM, CD34 and CD45. PGE2 content in tumor tissue was determined. Transcript of CD133 in tumor tissue was lower in patients treated with NSAIDs (0.28 ± 0.07 vs. 0.51 ± 0.08; p<0.03) as well as some other stem cell-related transcripts. In treated patients 36% of all tumors stained positive for CD133 compared to 71% in sham-treated control patients (p<0.05). Short survivors with Dukes' B tumors displayed 78% CD133 expression as compared to 33% of tumors in long-term survivors (p<0.002). Tumor tissue PGE(2) content was negatively related to patient survival. Our results show that short-term preoperative NSAID treatment downregulates colon cancer tissue expression of Prominin 1/CD133, a stem cell marker indicative of survival prognosis as confirmed.

The Extra Domain A of Fibronectin Increases VEGF-C Expression in Colorectal Carcinoma Involving the PI3K/AKT Signaling Pathway

The extra domain A (EDA)-containing fibronectin (EDA-FN), an alternatively spliced form of the extracellular matrix protein fibronectin, is predominantly expressed in various malignancies but not in normal tissues. In the present study, we investigated the potential pro-lymphangiogenesis effects of extra domain A (EDA)-mediated vascular endothelial growth factor-C (VEGF-C) secretion in colorectal carcinoma (CRC). We detected the expressions of EDA and VEGF-C in 52 human colorectal tumor tissues and their surrounding mucosae by immunohistochemical analysis, and further tested the correlation between the expressions of these two proteins in aforementioned CRC tissues. Both EDA and VEGF-C were abundantly expressed in the specimens of human CRC tissues. And VEGF-C was associated with increased expression of EDA in human CRC according to linear regression analysis. Besides, EDA expression was significantly correlated with lymph node metastasis, tumor differentiation and clinical stage by clinicopathological analysis of tissue microarrays containing tumor tissues of 115 CRC patients. Then, human CRC cell SW480 was transfected with lentivectors to elicit expression of shRNA against EDA (shRNA-EDA), and SW620 was transfected with a lentiviral vector to overexpress EDA (pGC-FU-EDA), respectively. We confirmed that VEGF-C was upregulated in EDA-overexpressed cells, and downregulated in shRNA-EDA cells. Moreover, a PI3K-dependent signaling pathway was found to be involved in EDA-mediated VEGF-C secretion. The in vivo result demonstrated that EDA could promote tumor growth and tumor-induced lymphangiogenesis in mouse xenograft models. Our findings provide evidence that EDA could play a role in tumor-induced lymphangiogenesis via upregulating autocrine secretion of VEGF-C in colorectal cancer, which is associated with the PI3K/Akt-dependent pathway.