Introduction

Colorectal cancer (CRC) is a global health concern, with high mortality rates and limited success in early detection and effective treatment of metastatic disease. While conventional treatments such as surgery, chemotherapy, and radiation therapy have shown some success, there is still a need for new and more effective treatment options. Repurposing existing drugs for cancer treatment has gained increasing attention due to its cost-effective nature and the availability of a large number of approved drugs with established safety profiles. In this study, we investigate the potential of using metformin and celecoxib as adjuvant agents for CRC treatment. Both drugs have been shown to have anti-cancer properties and are well-established for the treatment of other conditions. By analyzing gene expression data sets for these drugs and performing in vitro assays, we aim to evaluate their effectiveness as combination therapy and explore the underlying mechanisms of action.

Aims & Methods

Gene expression data sets for these drugs were obtained from the GEO database (GSE11237 and GSE67342) and analyzed using the R Limma package. Fold-change (FC; value > 1) and p-value ≤ 0.05 are considered as DEG selection criteria. MTT assay, apoptosis, and cell cycle analysis will be performed to evaluate the cytotoxic effect of these drugs on CaCO2 and SW48 cell lines. Furthermore, to assess the expression of the candidate genes identified in the in-silico phase, we will perform real-time PCR analysis.

Results

After performing gene expression analysis using the R Limma package, we found that only one gene, ID1, was identified as commonly expressed in both datasets. STRING server was used to find the intermediate network between ID1 and other human genes. Finally, the resulting gene network was determined by using Gephi software. Based on this, three modules were obtained. The genes in the module containing ID1 are MYOD1, SMAD4, SMAD9, SMAD5, and TCF3. Among the resulting genes, considering that ID1 is a hub gene and the functional pathway of this gene is TGFB. We selected the SMAD4 gene, which plays a role in this pathway, as a companion gene to determine the expression level of these two genes in treatment with both drugs.
Our results suggest a potential functional role for ID1 in the cellular response to treatment with the drugs metformin and celecoxib in CRC.

Conclusion

In conclusion, our study provides evidence that the combination therapy of metformin and celecoxib may have the potential as adjuvant therapy for CRC. These findings contribute to the understanding of the molecular mechanisms underlying the therapeutic effects of these drugs in CRC and provide a basis for the development of new adjuvant therapies that could improve patient outcomes and reduce healthcare costs.

Introduction

Colorectal cancer (CRC) screening is well established worldwide. However, the suboptimal sensitivity of the Fecal Immunochemical Test (FIT) for precancerous lesions justifies the search for new tools, namely microRNAs (miRs). In recent years, the detection of these molecules in liquid biopsies has gained increasing interest, as they reflect CRC progression and enable the early detection of precancerous lesions. Moreover, using miRs as a blood-based screening test could potentially increase patient adherence.

Aims & Methods

In this context, we aimed to identify circulating miR profiles for the early detection of CRC. Based on our previous literature review, we selected four miRs (miR-21-5p, miR-92a-3p, miR-451a and miR-199a-5p) with the best performance for the detection of CRC and high-grade dysplasia lesions (HGD). MiR expression was evaluated using RT-qPCR in plasma samples from 80 individuals who underwent colonoscopy with different clinical outcomes, namely, individuals without lesions found on colonoscopy, low-grade dysplasia lesions, high-grade dysplasia lesions, and patients diagnosed with CRC.

Results

Our findings demonstrate a decreased expression of all miRs in CRC and HGD lesions. Regarding HGD identification, miR-21-5p demonstrate a sensitivity of 68% (95% CI: 46 – 85%) and a specificity of 100% (95% CI: 84 – 100). Moreover, miR-92a-3p, miR-451a and miR-199a-5p demonstrated a sensitivity higher than 85% for HGD lesions detection, despite low sensitivity (around 70%). Concerning CRC detection, miR-92a-3p and miR-199a-5p demonstrated sensitivity of approximately 65%, with specificities of 70%, for miR-92a-3p (95% CI: 48 – 85%), and 80% for miR-199a-5p (95% CI: 58 – 92%). Furthermore, miR-21-5p showed outstanding performance in CRC identification with a sensitivity of 95% (95% CI: 75 – 100%), for an AUC of 0.900 (95% CI: 0.807 – 0.993). Moreover, when a negative result was obtained for one of these miRs, the presence of colorectal cancer lesions could be accurately excluded (negative predictive value: 98%).

Conclusion

These findings suggest that circulating miRs have the potential to improve the detection of precancerous lesions, surpassing the performance of FIT. Moreover, given their accuracy in identifying individuals without lesions, they could serve as effective confirmatory screening tools, reducing the number of unnecessary colonoscopies.
​

Introduction

Colorectal cancer (CRC) remains a significant public health concern, with its incidence steadily rising worldwide. Despite notable strides in treatment modalities, predicting disease progression and tailoring therapeutic approaches remains a challenge. In this context, identifying reliable and simple prognostic factors is crucial for optimizing disease outcomes.

Aims & Methods

The aim of this study was to investigate the predictive and prognostic value of C-reactive protein-to-albumin ratio (CAR) in CRC. We conducted a retrospective and descriptive study including patients diagnosed with primary colon or rectal cancer, over an 8-year period [January 2014-June 2022]. Patient demographics, inflammatory markers, tumor characteristics, treatment and follow-up data were collected. Baseline CAR was calculated for each patient. Data were entered and analyzed by SPSS software version 26.

Results

Sixty-five patients were included, with a mean age at disease diagnosis of 61 ± 14 years and a sex ratio M/F=1.82. The median level of CAR was 0.4 [0.1-1.3]. No significant association was noted of CAR neither with age of disease onset (p=0.26), nor gender (p=0.86), nor smoking (p=0.88) nor family history of CRC (p= 0.54). For biological data, high levels of carcinoembryonic antigen and CA19-9 were significantly associated with a higher level of CAR (p=0.002 and p=0.01 respectively). We noted a significant association between lower levels of CAR and hematochezia (p=0.01) and so, rectal tumors were associated with lower levels of CAR compared with colonic tumors (26.2 vs 36; p=0.05). Circumferential, stenoting and friable tumors with hemorrhagic appearance were significantly associated with higher CAR levels (p=0.01; p=0.001 and p=0.01 respectively). We noted a significant association between high levels of CAR and metastases (50.1 vs 29.8; p=0.002) and mainly hepatic metastases (p=0.004). Regarding treatment, neither primary surgery nor urgent surgery nor adjuvant chemotherapy were associated with CAR (p=0.11; p=0.12 and p=0.85 respectively). However, an increased CAR was significantly associated with early readmission within 30 days subsequent to surgery (41.3 vs 26.7; p=0.01). Lower overall survival was associated with higher CAR rates (38.8 vs 29; p=0.04). Area under the receiver operating characteristic curve analysis showed that higher CAR was predictive of a shorter progression-free survival (AUROC = 0.65). The optimal cutoff value was 0.3 (sensitivity, 70%; specificity, 62%). A total of 36 (55%) patients had a CAR above this threshold. These patients presented more frequently with lymphopenia (p=0.03) and higher carcinoembryonic antigen rates (p=0.02). We noted no association between advanced TNM stage 3 or 4 and CAR levels, regardless of this threshold (p=0.36), and overall disease recurrence was similar for CAR above and below 0.3 (p=0.46).

Conclusion

C-reactive protein-to-albumin ratio is a strong predictor of progression-free survival and is associated with clinicopathological factors in CRC. It serves as a valuable tool for straightforward prognostic assessment, aiding in more personalized treatment decisions.

Introduction

Colorectal cancer (CRC) is the third most diagnosed cancer worldwide¹, with a similar prevalence observed in Denmark, where it ranks as the third most frequent malignancy². In recent decades, the incidence of early-onset colorectal cancer—defined as CRC diagnosed in individuals under 50 years of age—has been rising^3,4. Since CRC is often asymptomatic in the early stages, late-stage diagnosis is associated with high mortality rates⁵. Therefore, it is crucial to develop new strategies to identify risk factors and early tumour formation, enabling timely prevention, treatment, and improved patient outcomes⁶. While colonoscopy remains the gold standard, it is invasive and often unnecessary; data from Region Zealand show that nearly 60% of individuals undergoing colonoscopy after a positive faecal immunochemical test (FIT) do not have lesions requiring treatment⁷. The development of molecular diagnostics, such as faecal DNA-based screening, offers a promising alternative that could significantly improve early detection while reducing the number of unnecessary colonoscopies, particularly in high-risk populations such as those with inflammatory bowel disease (IBD), previous CRC history, or genetic predispositions.

Aims & Methods

We evaluated two alternative approaches for detecting colorectal cancer-related mutations from faecal-derived human DNA: short-read next-generation sequencing (NGS) and digital PCR (dPCR). The aim was to assess whether these methods could support a non-invasive screening strategy that does not rely on the detection of occult blood. Faecal samples were collected from patients in Region Zealand who tested positive in the Danish national colorectal cancer (CRC) screening programme between January 2022 and January 2023 (FIT ≥100 ng/mL).
Faecal DNA was extracted using the QIAamp Fast DNA Stool Mini Kit (Qiagen) with RNase treatment. Formalin-fixed paraffin-embedded (FFPE) tumour samples were processed with the GeneRead DNA FFPE Kit (Qiagen). For next-generation sequencing (NGS), libraries were prepared with the QIAseq Targeted DNA Pro kit (Qiagen), targeting regions commonly mutated in CRC. Digital PCR (dPCR) was performed using the QIAcuity Probe PCR Kit and a panel targeting six validated single-nucleotide variants (SNVs) in CRC driver genes.

Results

Of the 71 sequencing DNA libraries generated from faecal samples, only three yielded more than 100 high-quality human reads, highlighting the challenge of recovering host DNA from stool material due to high gut microbiome content and inhibitory substances. Across the 33 patients successfully analysed, a total of 163 faecal DNA variants were detected by either NGS or dPCR. Of these, 21 were classified as oncogenic or likely oncogenic in colorectal cancer, and five were pathogenic variants not directly linked to CRC. These combined results demonstrate the feasibility of faecal DNA-based mutation detection as a non-invasive screening tool and underscore the importance of adequate DNA recovery for reliable variant identification.

Conclusion

Our study provides compelling evidence that mutation-based faecal DNA screening is feasible and may augment current CRC detection strategies. While current sensitivity remains suboptimal, the high PPV of dPCR under stringent quality control is encouraging. Our findings contribute to a growing body of literature supporting the transition toward precision diagnostics in cancer screening. When implemented within a personalised framework, faecal DNA mutation analysis may reduce unnecessary colonoscopies and enable early detection of high-risk lesions.

References

1. Sung, H., et al. (2021). Global Cancer Statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA: A Cancer Journal for Clinicians 71(3), 209–249. DOI: 10.3322/caac.21660.
2. Njor, S. H., et al. (2018). Three years of colorectal cancer screening in Denmark. Cancer Epidemiology 57, 39–44. DOI: 10.1016/j.canep.2018.09.003.
3. Vuik, F. E., et al. (2019). Increasing incidence of colorectal cancer in young adults in Europe over the last 25 years. Gut 68(10), 1820–1826. DOI: 10.1136/gutjnl-2018-317592.
4. Akimoto, N., et al. (2021). Rising incidence of early-onset colorectal cancer – a call to action. Nature Reviews Clinical Oncology 18(4), 230–243. DOI: 10.1038/s41571-020-00445-1.
5. Ilyas, M. I. M. (2023). Epidemiology of Stage IV Colorectal Cancer: Trends in the Incidence, Prevalence, Age Distribution, and Impact on Life Span. Clinics in Colon and Rectal Surgery 37(2), 57–61. DOI: 10.1055/s-0043-1761447.
6. Wolf, A. M. D., et al. (2018). Colorectal cancer screening for average-risk adults: 2018 guideline update from the American Cancer Society. CA: A Cancer Journal for Clinicians 68(4), 250–281. DOI: 10.3322/caac.21457.
7. Nielson, C. M., et al. (2018). Positive predictive values of fecal immunochemical tests used in the STOP CRC pragmatic trial. Cancer Medicine 7(9), 4781–4790. DOI: 10.1002/cam4.1727.

Disclosure

The authors declare no competing interests.