Expression In Colorectal Tissues Research Articles

Wnt5a-induced M2 polarization of tumor-associated macrophages via IL-10 promotes colorectal cancer progression

BackgroundTumor-associated macrophages (TAMs) in the tumor microenvironment influence tumor initiation, invasion and metastasis. Several studies have shown that Wnt5a is mainly expressed in the tumor stroma, especially in TAMs. However, whether Wnt5a regulates the polarization and biological function of TAMs in colorectal cancer (CRC) is incompletely understood.MethodsImmunofluorescence staining was performed to detect CD68 and Wnt5a expression in colorectal tissues from patients (63 CRC specimens VS 20 normal tissues). RT-qPCR, flow cytometry, ELISA and inhibitors were carried out to explore the role of Wnt5a in the polarization of TAMs. Clone formation and transwell assays were performed to determine the effects of Wnt5a–treated macrophages on tumor proliferation, migration and invasion in vitro. Finally, a xenograft model was applied to confirm the effects of Wnt5a+ TAMs on CRC tumorigenesis.ResultsWe found that high Wnt5a+CD68+/CD68+ TAMs ratio was significantly associated with poor prognosis in CRC patients and Wnt5a+ TAM was an M2-like TAM subtype. Subsequently, we found that Wnt5a induced macrophages to secrete IL-10, which then acted as an autocrine cytokine to induce M2 polarization of these macrophages. IL-10 neutralizing antibody completely reversed the pro-M2 effect of Wnt5a. Mechanistically, the CaKMII-ERK1/2-STAT3 pathway was required for Wnt5a-mediated IL-10 expression in macrophages. Furthermore, Wnt5a-induced M2 macrophages promoted CRC cells proliferation, migration and invasion; knockdown of Wnt5a in TAMs significantly impaired the pro-tumor functions of TAMs.ConclusionsOur data indicate that Wnt5a could induce M2 polarization of TAMs by regulating CaKMII-ERK1/2-STAT3 pathway–mediated IL-10 secretion, ultimately promoting tumor growth and metastasis of CRC.

POM121 overexpression is related to a poor prognosis in colorectal cancer

ABSTRACTBackground: Nuclear pore membrane protein 121 (POM121) plays a crucial role in nucleocytoplasmic transport, but its significance in tumorigenesis and the progression of colorectal cancer (CRC) remains unknown. The aim of this study was to evaluate the relationship between POM121 and CRC.Methods: POM121 expression in colorectal tissues was analyzed at both the gene and protein levels. We investigated the connection between POM121 expression and clinicopathological features, as well as overall survival. A gene set enrichment analysis (GSEA) was performed, and a protein-protein interaction (PPI) network was constructed to determine the mechanism of POM121 in CRC.Results: CRC tissues displayed a striking increase in POM121 expression compared with colonitis and pericarcinomatous mucosa tissues (66.61% vs 24.36% vs 24.11%, respectively, p < 0.0167). POM121 overexpression was significantly associated with lymph node metastasis, distant metastasis, TNM stage, venous invasion, perineural invasion, preoperative CEA and CA19-9 levels, and Ki67 expression. CRC patients with high POM121 levels tended to have poor overall survival rates. POM121 may participate in the regulation of the cell cycle and DNA repair in CRC.Conclusions: Our results suggest that POM121 has the potential to serve as a novel prognostic biomarker in CRC patients.

Metastatic colorectal carcinomas with high SATB2 expression are associated with better prognosis and response to chemotherapy: a population-based Scandinavian study

Background: Survival and response to therapy in patients with metastatic colorectal cancer (mCRC) are very heterogeneous. There is an unmet need for better markers of prognosis and treatment benefit for mCRC patients. The homeobox 2 gene SATB2 has a highly specific expression in colorectal tissue and is associated with better prognosis in non-metastatic CRC.Material and methods: A population-based cohort of 798 mCRC patients was analysed. From primary tumour material, protein expression was assessed by immunohistochemistry. BRAF and KRAS mutation status was also determined. Associations with clinicopathological data, overall and progression-free survival and response to first-line chemotherapy were analysed.Results: Tumour tissue and clinical data were available from 467 patients. SATB2 was strongly expressed in 58% of cases, significantly more in left-sided, low-grade and wild-type BRAF tumours. Patients with high SATB2 tumours had longer overall survival compared with low SATB2 tumours (median 13 vs 8 months respectively, p < .001). Chemotherapy was given to 282 patients (63%). Patients with high SATB2 tumours had longer OS (median 22 vs 15 months respectively, p = .001) and more often responded to chemotherapy than those with low SATB2 (objective response 43% vs 29%, p = .02; clinical response 83% vs 67%, p = .004). Progression-free survival on first-line irinotecan chemotherapy was longer in high SATB2 cases (median 8 vs 4 months respectively, p = .019). Patients with both low SATB2 expression and mutated BRAF (n = 69) had particularly poor survival compared to the rest (median 8 and 12 months respectively, p = .001). In multivariable analysis, the SATB2 findings were independent of known clinicopathological prognostic markers, including BRAF mutation status.Conclusion: Patients with mCRC expressing high level of SATB2 have better prognosis and response to chemotherapy than those with low SATB2 expression. Patients with both low SATB2 expression and mutated BRAF had particularly poor prognosis and could thus benefit from more aggressive therapies.

Dickkopf-4 gene expression is associated with differentiation and lymph node metastasis in colorectal cancer.

Background and AimAlthough high expression of Dickkopf‐4 (DKK‐4) in colorectal cancer has been reported in previous studies, its impact on clinicopathological features, including the prognosis and mechanism of expression, has not been well clarified to date.Methods(i) DKK‐4 protein expression was analyzed by immunohistochemical staining with anti‐DKK‐4 antibody using archived formalin‐fixed paraffin‐embedded specimens obtained at surgery from 122 patients with colorectal cancer, and its association with clinicopathological findings was also investigated. (ii) The association between intratumoral DKK‐4 protein expression and somatic hotspot mutations in cancer‐associated genes in 40 patients was investigated using a next‐generation sequencer.ResultsIn cross‐section, DKK‐4 protein expression in colorectal tissue was related to an adenocarcinoma of a histologically differentiated type (tub1/tub2, P = 0.032) and distant metastasis (P = 0.012). Longitudinally, however, DKK‐4 was not an independent prognostic factor determining overall survival (OS). On the other hand, in patients with metastasis, high DKK‐4 protein was independently associated with short OS (P = 0.013). In addition, colorectal cancer tissue with high DKK‐4 protein expression was associated with hotspot mutations in Wnt/β catenin‐signaling molecules (APC, CTNNB1, and FBXW7, P = 0.03).ConclusionIntratumoral DKK‐4 expression, by partly reflecting the Wnt/β catenin pathway, is strongly associated with the advancement of colorectal cancer and OS in colorectal cancer patients with metastasis, although further studies are needed.

Diet and lifestyle factors associated with miRNA expression in colorectal tissue.

MicroRNAs (miRNAs) are small non-protein-coding RNA molecules that regulate gene expression. Diet and lifestyle factors have been hypothesized to be involved in the regulation of miRNA expression. In this study it was hypothesized that diet and lifestyle factors are associated with miRNA expression. Data from 1,447 cases of colorectal cancer to evaluate 34 diet and lifestyle variables using miRNA expression in normal colorectal mucosa as well as for differential expression between paired carcinoma and normal tissue were used. miRNA data were obtained using an Agilent platform. Multiple comparisons were adjusted for using the false discovery rate q-value. There were 250 miRNAs differentially expressed between carcinoma and normal colonic tissue by level of carbohydrate intake and 198 miRNAs differentially expressed by the level of sucrose intake. Of these miRNAs, 166 miRNAs were differentially expressed for both carbohydrate intake and sucrose intake. Ninety-nine miRNAs were differentially expressed by the level of whole grain intake in normal colonic mucosa. Level of oxidative balance score was associated with 137 differentially expressed miRNAs between carcinoma and paired normal rectal mucosa. Additionally, 135 miRNAs were differentially expressed in colon tissue based on recent NSAID use. Other dietary factors, body mass index, waist and hip circumference, and long-term physical activity levels did not alter miRNA expression after adjustment for multiple comparisons. These results suggest that diet and lifestyle factors regulate miRNA level. They provide additional support for the influence of carbohydrate, sucrose, whole grains, NSAIDs, and oxidative balance score on colorectal cancer risk.

PTU-255 Characterisation of drug transporter gene expression in colorectal tissue and cell lines: induction with anti-retrovirals for microbicide optimisation

Introduction Drug transporter expression in the colorectal epithelium is likely to play a role in the mucosal disposition of anti-retrovirals (ARVs) in rectal microbicidal preparations and impact their efficacy in prevention of HIV-1 infection. However, there is limited information on expression levels available. This study aims to assess expression of 84 drug transporter genes in human colorectal tissue and representative cell lines pre and post ARV exposure. Method Drug transporter mRNA expression was quantified from colorectal biopsies (n = 12) and 6 colorectal cell lines using real time PCR. Relative mRNA expression was quantified in Caco-2 cells after induction with tenofovir (TFV; 5 mM), dapivirine (DPV; 10 µM) and darunavir (DRV; 250 µM) for up to 3 days. Data were analysed using Pearson’s correlation (r), hierarchical clustering and principal component analysis. Results Fifty-eight of the 84 transporters were expressed in colorectal tissue. SLC28A2/CNT2 was the most expressed uptake transporter (>25 fold increase) followed by efflux transporters ABCB1/P-gp and ABCC3/MRP3 (4–5 fold increase). No difference was noted between individual patients, either sexes or biopsy sites (rectum or sigmoid) (r = 0.95–0.99). Similarities between tissue and cell lines were low (r values 0.67–0.81). Principal component analysis showed distinct clustering of colorectal tissue and cell lines with Caco-2 cells showing least variance with tissues. Immunohistochemical staining of 7 colorectal biopsy sections confirmed high expression of Pgp and CNT2 drug transporters. TFV stimulation of Caco-2 cells resulted in > 2 fold increase in expression of MRP5, LAT2, OATPE and GLUT1 mRNA. DPV stimulation resulted in > 2 fold increase in expression of GLUT1, PEPT1 and ABCA1. DRV stimulation resulted in > 2 fold increase in expression of SLC3A2, OATPE and MCT3 mRNA. Conclusion This study has enumerated drug transporter mRNA expression in colorectal tissue and cell lines. There was partial correlation between cell lines and tissue limiting cell line use as in vivo surrogate. TFV and DPV did not induce majority of efflux transporters in Caco-2 cells. This would entail increased retention of drug within the mucosa and as a result enhanced delivery to CD4 T cells. Further studies will elucidate whether combination of ARVs will be similarly effective. Disclosure of interest None Declared.

Characterisation of Drug Transporter Gene Expression in Colorectal Tissue and Cell Lines: Induction with Anti-retrovirals for Microbicide Optimization

Characterisation of Drug Transporter Gene Expression in Colorectal Tissue and Cell Lines: Induction with Anti-retrovirals for Microbicide Optimization

Expression and localization of apolipoprotein M in human colorectal tissues

BackgroundIt has been well documented that apolipoprotein M (apoM) is principally expressed in the liver and kidney. However we found that there was weak apoM expression in other tissues or organs too, which could not be ignored. In the present study, we therefore examined apoM expression in human colorectal tissues including cancer tissues, cancer adjacent normal tissues, polyp tissues and normal mucosa as well as inflammatory mucosa.MethodsTissue samples were collected from patients who underwent surgical resection or endoscopic examination. ApoM mRNA levels were determined by the real-time RT-PCR and apoM protein mass were examined by the immunohistochemistry.ResultsApoM protein can be detected in all colorectal tissues. However, apoM protein mass were significantly lower in the cancer tissues than its matched adjacent normal tissues, polyp tissues, normal mucosa and inflammatory mucosa. In parallel, apoM mRNA levels in the colorectal cancer tissues (0.0536 ± 0.0131) were also significantly lower than those in their adjacent normal tissues (0.1907 ± 0.0563) (P = 0.033). Interestingly, apoM mRNA levels in colorectal cancer tissues were statistic significant higher in the patients with lymph node metastasis than the patients without lymph node metastasis (P = 0.008). Patients under Dukes' C and D stages had much higher apoM mRNA levels than patients under Dukes' A and B stages (P = 0.034).ConclusionIt is concluded that apoM could also be expressed in human colorectal tissues besides liver and kidney. ApoM mRNA levels in the colorectal cancer tissues were significantly increased in the patients with lymph node metastasis. Whether increased apoM expression in the patients with lymph node metastasis being related to patients' prognosis and the physiopathological importance of apoM expression in colorectal tissues need further investigation.

Clinicopathologic significance of survivin expression in colorectal adenocarcinoma

ABSTRACT Background and aim: Survivin, an antiapoptotic protein, is involved in both the control of the cell division and the inhibition of apoptosis. The regulation of apoptotic cell death may have a profound effect on the carcinogenesis and progression of colorectal cancer. This study was designed to evaluate the survivin expression in colorectal tissues and to investigate whether the survivin expression is associated with the clinicopathologic characteristics and the patient survival. Methods: Immunohistochemical analysis of survivin was performed using the tissue microarrays of 622 colorectal tissue samples from 529 colorectal adenocarcinomas, 40 normal mucosa, 34 adenomatous polyps, and 59 metastatic lymph nodes. Results: Normal colorectal mucosa were completely negative for survivin (0/40, 0%), but survivin was expressed in adenomatous polyps (8/34, 23.5%), adenocarcinomas (327/529, 61.8%), and metastatic lymph nodes (27/59, 45.8%). In the analyses between the survivin expression and clinicopathologic parameters, the survivin expression was correlated with tumor location in colon (P= 0.026), lymph node metastasis (P= 0.003) and American Joint Committee on Cancer (AJCC) stage (P= 0.040). The patient survival was significantly associated with age, histologic grade, AJCC stage, and lymphovascular invasion. On stage wise analyses of patient survival, the survivin expression was significantly correlated with overall survival (P &lt; 0.001) and disease free survival (P &lt; 0.001), especially in stage II colorectal adenocarcinomas. Conclusions: The results suggest that the survivin expression is a relatively frequent early event of colorectal carcinogenesis and may play an important role in the adenoma‐carcinoma transition sequence. Survivin expression provides important prognostic information and an apoptosis‐based therapeutic target in colorectal adenocarcinomas.

Immunohistochemical expression of folate receptor α in colorectal carcinoma: patterns and biological significance

Folate receptor alpha (FRalpha) has emerged as a potential cancer therapy target with several folate-linked therapeutic agents currently undergoing clinical trials. In addition, FRalpha expression in tumors may offer prognostic significance. Most studies on FRalpha expression used reverse transcriptase polymerase chain reaction or cytofluorimetric assays. The applicability of such methods to paraffin-embedded tissues is limited. The aims of this study were to assess the feasibility of immunohistochemistry in detecting FRalpha expression and to assess the patterns and clinical significance of FRalpha expression in colorectal tissues. We used tissue microarrays containing 152 normal colorectal mucosa samples, 42 adenomas, 177 primary, and 52 metastatic colorectal carcinomas. Our results showed that staining for FRalpha on colorectal tissues was simple and easy to read. FRalpha positivity was more frequent in carcinomas (33% in primaries and 44% in metastases) than in normal mucosa or adenoma (7% in both) (P < .001). Positive staining in primary carcinomas correlated with younger age (n = 130) (P = .008), presence of distant metastasis (n = 130) (P = .043), and non-high-frequency microsatellite instability status (as detected by the standard polymerase chain reaction method using the 5 National Cancer Institute-recommended markers) (n = 77) (P = .006). Positive staining in primary carcinomas also correlated with a worse 5-year disease-specific survival (P = .04) on univariate but not multivariate analysis. Thus, our data show that there is selective expression of FRalpha in some colorectal cancers, providing a foundation for investigating the use of folate conjugates for imaging and therapy of colorectal tumors. Furthermore, our results suggest that a possible association exists between FRalpha expression and the microsatellite instability status in colorectal carcinoma. The significance of such an association as well as the prognostic value of FRalpha expression deserves further exploration.

Expression of Cathepsin D in Colorectal Adenocarcinomas: Correlation with Clinicopathologic Features

Expression of Cathepsin D in Colorectal Adenocarcinomas: Correlation with Clinicopathologic Features

Expression of 15-lipoxygenase-1 is regulated by histone acetylation in human colorectal carcinoma.

15-Lipoxygenase-1 (15-LO-1) is expressed at higher levels in human colorectal tumors compared with normal tissue. 15-LO-1 is expressed in cultured human colorectal cells, but only after treatment with sodium butyrate (NaBT), which also stimulates apoptosis and cell differentiation. We examined the regulation of 15-LO-1 in human tissue and the colorectal carcinoma cell lines Caco-2 and SW-480 by treatment with histone deacetylase (HDAC) inhibitors: NaBT, trichostatin A (TSA) and HC toxin. Northern and western analysis showed that expression of 15-LO-1 was up-regulated by these HDAC inhibitors. Furthermore, HDAC inhibitors stimulated promoter activity of the 15-LO-1 gene approximately 12-to 21-fold using the -331/-23 region of the 15-LO-1 promoter, as measured with a luciferase-15-LO-1 promoter-reporter system, suggesting that 15-LO-1 is regulated at the transcriptional level by HDAC inhibitors. Histone proteins in colorectal cells were acetylated after treatment with HDAC inhibitors. Histone acetylation was also measured in human colorectal tissue and a correlation was observed between increased histone acetylation and 15-LO-1 expression. Thus, regulation of 15-LO-1 expression in colorectal tissues appears to occur by a novel and new mechanism associated with histone acetylation. Moreover, these results suggest that 15-LO-1 is a marker that reflects histone acetylation in colorectal carcinoma.

Biochemistry and pathological importance of mucin-associated antigens in gastrointestinal neoplasia

Publisher Summary This chapter focuses on mechanisms of alterations of Mucin-Associated Antigens in gastrointestinal neoplasia and their implications in clinical diagnostic and therapeutic oncology. The chapter focuses on real mucins (MUC), taking MUC1 and MUC2 as models. In addition, the chapter defines mucins and discusses the biochemistry and molecular biology of mucins, such as structural aspects of mucins, biosynthesis of mucins, glycosylation site specificity and biosynthetic routes of O-glycosylation, and antigenicity of mucins. During carcinogenesis, alterations of mucin peptide and mucin glycosylation occur. Although glycosylation of proteins should be dependent on the rate of synthesis, and overexpression of mucin species in certain cancers is associated with reduced glycosylation, there is no clear-cut evidence for a causal relationship between these events. The chapter provides an account for the expression of mucin-asssociated antigens in gastric tissues (in non-neoplastic and neoplastic gastric tissues); and expression in colorectal tissues (in non-neoplastic colorectal tissues and colorectal adenoma-carcinoma sequence). Mucin-associated peptide and carbohydrate antigens play an important role in different aspects of proliferation, local invasion, and metastasis of cancer cells. Thus, the carbohydrate antigens gained clinical importance as prognostic markers in a great variety of human carcinomas. Serum tumor markers (circulating antigens) also help in clinical tumor diagnostics. The chapter concludes with the perspectives in tumor immunology and cancer therapy. Mucin peptide cores, in addition to substituting carbohydrate structures, are involved in different types of cellular and humoral immune response in cancer. Biosynthesized glycopeptides containing clustered saccharide domains and peptides that mimic carbohydrates may be applied in future studies on therapy of gastrointestinal carcinomas.