Loss of Smad4 protein expression and 18qLOH as molecular markers indicating lymph node metastasis in colorectal cancer-a study matched for tumor depth and pathology

Current researches have proposed a genetic model for colorectal cancer (CRC), in which the sequential accumulation of mutations in specific cancer-related genes, including adenomatous polyposis coli (APC), K- ras, and p53, drives the transition from normal epithelium through increasing adenomatous dysplasia to colorectal cancer. To identify patients with an increased risk of tumor recurrence or metastasis and evaluate the prognostic values of APC, K- ras, and p53 gene mutations, we investigated the frequency of these three mutated genes in tumors and sera of CRC patients. APC, K- ras, and p53 gene mutations in primary tumor tissues and their paired preoperative serum samples of 118 CRC patients were detected by using polymerase chain reaction– single strand conformation polymorphism (PCR-SSCP) analysis, followed by direct DNA sequencing of the PCR-amplified genomic DNA. Subsequently, serum molecular markers were analyzed for their correlation with patients’ clinicopathologic features and presence of postoperative recurrence/metastasis. We did not observe any significant difference in the association of APC or K- ras or p53 gene mutations in primary tumors with patients’ demographic data (all were P > 0.05). In contrast, both serum APC and p53 molecular markers were closely correlated with lymph node metastasis and TNM stage (both P < 0.05). Moreover, the serum overall molecular markers (at least one of the three markers) were prominently associated with depth of tumor invasion ( P = 0.033), lymph node metastasis ( P < 0.001), and TNM stage ( P < 0.001). In addition, a significantly higher postoperative metastasis/recurrence rate in patients positive for overall molecular markers compared to those negative for these molecular markers were also demonstrated ( P < 0.001). APC and K- ras molecular markers were more frequently observed in patients with locoregional metastasis (both P < 0.05), while p53 molecular marker was usually detected in the cases of peritoneal metastasis ( P = 0.004). Our findings suggest that serum molecular markers are potentially useful in the determination of colorectal cancer patients harboring gene mutations at high risk of metastasis. Serial analysis is warranted in order to assess their long-term prognostic significance and the therapeutic implications.

Download Full-text

Loss of ASXL1 protein is associated with lymph node metastasis in colorectal cancer

10.21203/rs.2.14131/v1 ◽

2019 ◽

Author(s):

Jun Ho Lee ◽

Ju-Hee Lee ◽

Byung Kyu Ahn ◽

Seung Sam Paik ◽

Hyunsung Kim ◽

...

Keyword(s):

Colorectal Cancer ◽

Lymph Node ◽

Lymph Node Metastasis ◽

Protein Expression ◽

Lymph Nodes ◽

Disease Free Survival ◽

Prognostic Impact ◽

Metastatic Lymph Nodes ◽

Node Metastasis ◽

Metastatic Lymph

Abstract Background The function of ASXL1 protein in colorectal cancer has not been investigated yet. The purpose of this study was to investigate the clinicopathological and prognostic impact of ASXL1 protein expression on colorectal cancer.Methods We performed immunohistochemical staining of ASXL1 protein using tissue microarrays of 408 colorectal cancers, 46 normal colonic mucosae, 48 adenomas, and 92 metastatic lymph nodes. The intensity of expression was scored as 0–3, and the extent of staining was scored as 0–4, based on the percentage of positive cells. The immunoreactivity score (IRS) was calculated by multiplying the two scores.Results ASXL1 protein expression rates were 89.1% in normal mucosae, 72.9% in tubular adenomas, 44.4% in adenocarcinomas, and 28.3% in metastatic lymph nodes ( p < 0.001). With respect to the IRS cut-off score, the mean tumor size was smaller in the IRS 0–6 group than in the IRS 8–12 group (4.9 ± 2.1 vs. 6.3 ± 2.7 cm, p = 0.002). Lymph node metastasis was more frequent in the IRS 0–6 group than in the IRS 8–12 group (56.3% vs. 33.3%, p = 0.034). Lymphatic invasion was more frequent in the 0–6 group than in the IRS 8–12 group (56.0% vs. 33.3%, p = 0.035). The 5-year disease-free survival rate did not differ between two groups at stage II and stage III.Conclusions ASXL1 protein might act as a tumor suppressor in colorectal cancer. The loss of ASXL1 expression might be associated with metastasis via the lymphatic system to the lymph nodes.

Download Full-text

High Level of ASXL1 Protein Is Associated With Better Disease-Free Survival in Colorectal Cancer

Journal of Global Oncology ◽

10.1200/jgo.18.80800 ◽

2018 ◽

Vol 4 (Supplement 2) ◽

pp. 200s-200s

Author(s):

K. Lee

Keyword(s):

Colorectal Cancer ◽

Lymph Node ◽

Lymph Node Metastasis ◽

Protein Expression ◽

Disease Free Survival ◽

Colorectal Cancers ◽

Free Survival ◽

Node Metastasis ◽

Positive Expression ◽

Disease Free

Background: ASXL1 gene is on chromosome region 20q11.21. Either amplification in cervical cancer or truncation mutations in colorectal cancers with microsatellite instability (MSI), malignant myeloid diseases, chronic lymphocytic leukemia, liver, prostate and breast cancers occurred. The functional and the prognostic roles of ASXL1 mutations and the expression of protein in colorectal cancer are still unknown. Aim: The aim of this study is to investigate the functional roles of ASXL1 mutations and the expression of protein in colorectal cancer. Methods: We performed NGS of 10 colorectal cancer with peritoneal seeding to find genetic markers for aggressive phenotype. All showed a frameshift deletion at codon 1934delG. To clinically validate the functional and the prognostic roles of the mutations, we performed an immunohistochemical staining (IHC) on tissue microarrays of 414 consecutive colorectal cancers. Results: The ASXL1 protein expression was strong positive in 5.8% (24 patients), moderate positive in 38.5% (157 patients) and negative in 55.6% (227 patients). The patients with negative ASXL1 expression had more lymph node metastasis than the patients with strong positive expression [59.0% (134/227 patients) vs 33.3% (8/24 patients), P = 0.038]. None of the patients with strong positive expression had recurrent disease in the stage I-III cancers [0% (0/21 patients) vs 19.4% (27/139 patients) vs 18.9% (34/180 patients)] and the disease-free survival rate of the patients with strong positive expression was significantly better than that of the patients with moderate positive or negative expression ( P = 0.037; P = 0.031). Conclusion: The decreased level of the expression of the ASXL1 protein was associated with lymph node metastasis in its progression of cancer. Strong positive ASXL1 protein expression was a 'good' prognostic factor of colorectal cancers. The ASXL1 protein might be tumor suppressive in colorectal cancer.

Download Full-text

Expression of TMEM16A in Colorectal Cancer and Its Correlation With Clinical and Pathological Parameters

Frontiers in Oncology ◽

10.3389/fonc.2021.652262 ◽

2021 ◽

Vol 11 ◽

Author(s):

Hongxia Li ◽

Qiwei Yang ◽

Sibo Huo ◽

Zhenwu Du ◽

Fei Wu ◽

...

Keyword(s):

Colorectal Cancer ◽

Lymph Node ◽

Lymph Node Metastasis ◽

Protein Expression ◽

Mrna Expression ◽

P53 Protein ◽

Clinicopathological Parameters ◽

Strong Positive Correlation ◽

Independent Predictive Factor ◽

Node Metastasis

TMEM16A is a recently identified calcium-activated chloride channel (CaCC) and its overexpression contributes to tumorigenesis and progression in several human malignancies. However, little is known about expression of TMEM16A and its clinical significance in colorectal cancer (CRC). TMEM16A mRNA expression was determined by quantitative real time-PCR (qRT-PCR) in 67 CRC tissues and 24 para-carcinoma tissues. TMEM16A protein expression was performed by immunohistochemistry in 80 CRC tissues. The correlation between TMEM16A expression and clinicopathological parameters, and known genes and proteins involved in CRC was analyzed. The results showed that TMEM16A mRNA expression was frequently detected in 51 CRC tissues (76%), whereas TMEM16A protein expression was determined at a relatively lower frequency (26%). TMEM16A mRNA expression in tumor tissues was higher than its expression in normal para-carcinoma tissues (P < 0.05). TMEM16A mRNA expression was significantly correlated with TNM stage (p = 0.039) and status of lymph node metastasis (p = 0.047). In addition, there was a strong positive correlation between TMEM16A mRNA expression and MSH2 protein. More importantly, TMEM16A protein expression was positively associated with KRAS mutation, and negatively correlated with mutant p53 protein. Logistic regression analysis demonstrated that TMEM16A mRNA expression was an important independent predictive factor of lymph node metastasis (OR = 16.38, CI: 1.91–140.27, p = 0.01). TMEM16A mRNA and protein expression was not significantly related with patient survival. Our findings provide original evidence demonstrating TMEM16A mRNA expression can be a novel predictive marker of lymph node metastasis and TMEM16A protein expression may be an important regulator of tumor proliferation and metastasis in CRC.

Download Full-text

RacGAP1 expression, increasing tumor malignant potential, as a predictive biomarker for lymph node metastasis and poor prognosis in colorectal cancer.

Journal of Clinical Oncology ◽

10.1200/jco.2015.33.3_suppl.555 ◽

2015 ◽

Vol 33 (3_suppl) ◽

pp. 555-555

Author(s):

Hiroki Imaoka ◽

Yuji Toiyama ◽

Susumu Saigusa ◽

Koichiro Mori ◽

Tomofumi Noguchi ◽

...

Keyword(s):

Colorectal Cancer ◽

Lymph Node ◽

Lymph Node Metastasis ◽

Protein Expression ◽

Clinical Significance ◽

Poor Prognosis ◽

Cellular Proliferation ◽

Migration And Invasion ◽

Node Metastasis ◽

Potential Biomarker

555 Background: Rac GTPase activating protein (RacGAP) 1 plays a key role in controlling various cellular phenomena including cytokinesis, transformation and migration. Recently, the clinical significance of RacGAP1 expression has been reported in several malignancies. However, direct association between the RacGAP1 expression and colorectal cancer (CRC) has not been fully investigated. The aim of this study is to elucidate the function and clinical significance of RacGAP1 expression in CRC. Methods: The intrinsic functions of RacGAP1 in CRC cells were analyzed using small interfering RNA (siRNA). We analyzed RacGAP1 mRNA expression in surgical specimens from 193 CRC patients (Cohort 1) by real-time polymerase chain reaction. Then, we validated RacGAP1 protein expression using formalin-fixed paraffin-embedded samples from 298 CRC patients (Cohort 2) by immunohistochemistry. Finally, we evaluated the association between RacGAP1 mRNA and protein expression and clinicopathological data. Results: Reduced RacGAP1 expression by siRNA in CRC cell lines showed significantly decreased cellular proliferation, migration, and invasion. In Cohort 1, RacGAP1 expression in CRC was significantly higher than in adjacent normal mucosa, and increased according to TNM stage progression. High RacGAP1 expression in tumors was significantly associated with progression and prognosis. In Cohort 2, RacGAP1 protein was overexpressed mainly in the nuclei of CRC cells; however, its expression was scarcely observed in normal colorectal mucosa. RacGAP1 protein expression was significantly higher in CRC patients with higher T stage, vessel invasion, and lymph node and distant metastasis. Increased expression of RacGAP1 protein was significantly associated with poor disease-free and overall survival. Multivariate analyses revealed that high RacGAP1 expression was an independent predictive marker for lymph node metastasis, recurrence, and poor prognosis in CRC. Conclusions: Our data provide novel evidence for the biological and clinical significance of RacGAP1 as a potential biomarker for identifying patients with lymph node metastasis and poor prognosis in CRC.

Download Full-text

Microenvironmental markers are correlated with lymph node metastasis in invasive submucosal colorectal cancer

Histopathology ◽

10.1111/his.14388 ◽

2021 ◽

Author(s):

Tamotsu Sugai ◽

Noriyuki Yamada ◽

Mitsumasa Osakabe ◽

Mai Hashimoto ◽

Noriyuki Uesugi ◽

...

Keyword(s):

Colorectal Cancer ◽

Lymph Node ◽

Lymph Node Metastasis ◽

Node Metastasis ◽

Submucosal Colorectal Cancer

Download Full-text

Increased Expression of VANGL1 is Predictive of Lymph Node Metastasis in Colorectal Cancer: Results from a 20-Gene Expression Signature

Journal of Personalized Medicine ◽

10.3390/jpm11020126 ◽

2021 ◽

Vol 11 (2) ◽

pp. 126

Author(s):

Noshad Peyravian ◽

Stefania Nobili ◽

Zahra Pezeshkian ◽

Meysam Olfatifar ◽

Afshin Moradi ◽

...

Keyword(s):

Gene Expression ◽

Colorectal Cancer ◽

Lymph Node ◽

Lymph Node Metastasis ◽

Candidate Genes ◽

Case Series ◽

Gene Expression Signature ◽

Gene Panel ◽

Expression Levels ◽

Node Metastasis

This study aimed at building a prognostic signature based on a candidate gene panel whose expression may be associated with lymph node metastasis (LNM), thus potentially able to predict colorectal cancer (CRC) progression and patient survival. The mRNA expression levels of 20 candidate genes were evaluated by RT-qPCR in cancer and normal mucosa formalin-fixed paraffin-embedded (FFPE) tissues of CRC patients. Receiver operating characteristic curves were used to evaluate the prognosis performance of our model by calculating the area under the curve (AUC) values corresponding to stage and metastasis. A total of 100 FFPE primary tumor tissues from stage I–IV CRC patients were collected and analyzed. Among the 20 candidate genes we studied, only the expression levels of VANGL1 significantly varied between patients with and without LNMs (p = 0.02). Additionally, the AUC value of the 20-gene panel was found to have the highest predictive performance (i.e., AUC = 79.84%) for LNMs compared with that of two subpanels including 5 and 10 genes. According to our results, VANGL1 gene expression levels are able to estimate LNMs in different stages of CRC. After a proper validation in a wider case series, the evaluation of VANGL1 gene expression and that of the 20-gene panel signature could help in the future in the prediction of CRC progression.

Download Full-text