SMAD4-201 transcript as a putative biomarker in colorectal cancer

Background: Transcripts with alternative 5'-untranslated regions (UTRs) result from the activity of alternative promoters and they can determine gene expression by influencing its stability and translational efficiency, thus executing complex regulation of developmental, physiological and pathological processes. Transcriptional regulation of human SMAD4, a key tumor suppressor deregulated in most gastrointestinal cancers, entails four alternative promoters. These promoters and alternative transcripts they generate remain unexplored as contributors to the SMAD4 deregulation in cancer. The aim of this study was to investigate the relative abundance of the transcript SMAD4-201 in colorectal cell lines and tissues in order to establish if its fluctuations may be associated with colorectal cancer (CRC). Methods: Relative abundance of SMAD4-201 in total SMAD4 mRNA was analyzed using quantitative PCR in a set of permanent human colon cell lines and tumor and corresponding healthy tissue samples from patients with CRC. Results: The relative abundance of SMAD4-201 in analyzed cell lines varied between 16 and 47%. A similar relative abundance of SMAD4-201 transcript was found in the majority of analyzed human tumor tissue samples, and it was averagely 20% lower in non-malignant in comparison to malignant tissue samples (p = 0.001). Transcript SMAD4-202 was not detectable in any of the analyzed samples, so the observed fluctuations in the composition of SMAD4 transcripts can be attributed to transcripts other than SMAD4-201 and SMAD4-202. Conclusion: The expression profile of SMAD4-201 in human tumor and non-tumor tissue samples may indicate the translational potential of this molecule in CRC, but further research is needed to clarify its usability as a potential biomarker for early diagnosis

Expression profile of CD81 gene transcripts in colorectal cancer

The activity profi le of alternative promoters may be an indicator of tumor characteristics. Alternative promoters of CD81 gene were shown to be differentially active in colon and rectal cancer tissue. The promoter active in colon and rectal cancer gives rise to transcripts CD81-205 and CD81-215, while the promoter active in normal gut mucosa gives rise to transcripts CD81-203 and CD81-213. This study aimed to analyze the relative abundance of the CD81 gene transcripts in colorectal cancer

Application of nanofat for treatment of traumatic faecal incontinence after sphincteroplasty – A pilot study

Aim: The aim of this study was to investigate whether the application of nanofat contain-ing stem cells improves continence in women who had previously undergone anal sphinc-teroplasty with unsatisfactory long-term outcomes.Method: This prospective pilot study included nine women with various degrees of anal incontinence who had previously undergone anal sphincteroplasty due to obstetric trauma. In all patients, the Wexner Incontinence Score (WS) and Faecal Incontinence Quality of Life Score (FIQLS), as well as anal manometry and endoanal ultrasound meas-urements, were performed before the procedure and during follow-up. In all patients, liposuction was performed and 50 ml of raw lipoaspirate was obtained and processed using a NanoFat Kit device. Approximately 20 ml of the mechanically emulsified and fil-trated fat was obtained and the anal sphincter complex was infiltrated with it. Patient fol-low-up was conducted in person or via telephone 6 and 12 months after the procedure.Results: The squeeze pressure was significantly increased 6 months after the proce-dure (p= 0.01). The external anal sphincter measured at the 12 o'clock position was sig-nificantly thicker (p= 0.04). A significant decrease in the WS was observed both 6 and 12 months after the procedure compared with baseline values (p< 0.05 for both).Conclusion: This study is the first to show that the application of nanofat as an inject-able product improves continence in patients with unsatisfactory results after sphinc-teroplasty, suggesting it to be a promising and effective therapeutic tool. The procedure is safe and can be easily performed as an ambulatory procedure

Analysis of the Prognostic Potential of Schlafen 11, Programmed Death Ligand 1, and Redox Status in Colorectal Cancer Patients

The Schlafen 11 (SLFN11) protein has recently emerged as pivotal in DNA damage conditions, with predictive potential for tumor response to cytotoxic chemotherapies. Recent discoveries also showed that the programmed death ligand 1 (PD-L1) protein can be found on malignant cells, providing an immune evasion mechanism exploited by different tumors. Additionally, excessive generation of free radicals, redox imbalance, and consequential DNA damage can affect intestinal cell homeostasis and lead to neoplastic transformation. Therefore, our study aimed to investigate the significance of SLFN11 and PD-L1 proteins and redox status parameters as prognostic biomarkers in CRC patients. This study included a total of 155 CRC patients. SLFN11 and PD-L1 serum levels were measured with ELISA and evaluated based on redox status parameters, sociodemographic and clinical characteristics, and survival. The following redox status parameters were investigated: spectrophotometrically measured superoxide dismutase (SOD), sulfhydryl (SH) groups, advanced oxidation protein products (AOPP), malondialdehyde (MDA), pro-oxidant–antioxidant balance (PAB), and superoxide anion (O2•–). The prooxidative score, antioxidative score, and OXY-SCORE were also calculated. The results showed significantly shorter survival in patients with higher OXY-SCOREs and higher levels of serum SLFN11, while only histopathology-analysis-related factors showed significant prognostic value. OXY-SCORE and SLFN11 levels may harbor prognostic potential in CRC patients

Long non-coding RNA H19 expression in rectal cancer and therapy response

Background & objectives: Long non coding RNA, H19 is an imprinted, maternally expressed gene, usually deregulated in diferent cancer types, including rectal cancer. This study aimed to investigate H19 role as a potential biomarker to predict therapy response in rectal cancer patients. Methods: The study included 14 patients diagnosed with rectal cancer, treated with neoadjuvant chemoradio therapy (nCRT). RNA was isolated by TRIzol reagent from samples of rectal cancer tissue before and after nCRT. Relative expression of H19 was normal- ized to housekeeping GAPDH gene, and expression was analysed by quantitative real-time PCR. Relative expression of H19 was calculated by 2-ΔCt method. Results: Relative expression of H19 was significantly increased in rectal cancer tissue after nCRT (0.244±0.408) compared to the tissue before nCRT (0.043±0.055), p=0.004, Wilcoxon test. According to tumour regression grade (TRG), 85.71% (12/14) of patients did not respond, while 14.28% (2/14) responded to pre- operative CRT. Responders (TRG1, TRG2) and non-responders (TRG3, TRG4) did not differ in H19 expression in tumour tissue before (p=0.659, Mann-Whitney U test) as well as after nCRT (p=0.999, Mann-Whitney U test). Receiver operating curve analy- sis indicates that H19 expression in colorectal tissue before nCRT can not be used as a biomarker for distinguishing responders from non-responders (AUC=0.625, 95%CI=0.257-0.992, p=0.583). Conclusion: Our study suggests H19 upregulation upon neoadju- vant chemoradiotherapy in rectal cancer. The potential predictive value of H19 as a biomarker of therapy response should be studied in a larger group of patients

Expression of long non-coding RNA HOTAIR in rectal cancer as a potential predictor of response to chemoradiotherapy

Patients with locally advanced rectal cancer are mainly treated with chemoradiotherapy (CRT) before surgery. Less than 20% of patients respond completely to neoadjuvant CRT. To avoid unnecessary treatment, biomarkers are being sought to identi fy pati ents with rectal cancer who do not respond to therapy. The HOX Transcript Anti sense Intergenic RNA (HOTAIR) is a long non-coding trans-acti ng RNA molecule that is frequently deregulated in cancers of the digestive tract and plays a role in chemoresistance. The aim of this study was to investigate HOTAIR as a potential biomarker for predicting treatment response in patients with rectal cancer

Genetic determinants of response to neoadjuvant chemoradiotherapy in locally advanced rectal cancer identified by whole exome sequencing

The cornerstone in the treatment of locally advanced rectal cancer (LARC) is neoadjuvant chemoradiotherapy (nCRT) followed by total mesorectal excision. Reliable predictors of response to nCRT in LARC remain an unmet need in colorectal cancer research. This study used high throughput DNA analysis to investigate genetic differences between highly responsive tumors and tumors resistant to nCRT.European Human Genetics Conference Hybrid Conference Glasgow, Scotland, UK JUNE 10–13, 202

Long non-coding RNA NEAT1 cannot be used as a diagnostic and prognostic biomarker in patients with locally advanced rectal cancer

Background & objectives: The NEAT1 (Nuclear Paraspeckle Assembly Transcript 1) gene encodes a long non-coding RNA that is deregulated in carcinomas of the gastrointestinal tract. Diagnostic and predictive potential of NEAT1 was investigated in patients with locally advanced rectal cancer. Methods: The study group consisted of 19 patients with rectal cancer treated with neoadjuvant chemoradiotherapy (nCRT). RNA was isolated with TRIzol reagent from samples of rectal cancer and noncancerous tissue before and after nCRT. The relative expression level of NEAT1 normalised to GAPDH was determined by qRT-PCR method. Results: Expression of NEAT1 did not difer between rectal cancer and noncancerous tissue before nCRT (p=0.953) and cancer and noncancerous tissue after nCRT (p=0.210). There was no diference in NEAT1 expression between tumour tissue before and after nCRT (p=0.079). NEAT1 was signifcantly higher in noncancerous tissue before than after nCRT (p=0.005). Therapy responders (TRG1, TRG2) and nonresponders (TRG3, TRG4) did not difer in NEAT1 levels in tumour tissue before (p=0.790) and after nCRT (p=0.352). NEAT1 expression in rectal cancer tissue before nCRT cannot be used as a biomarker to distinguish responders from non-responders (AUC=0.559, 95%CI=0- 1, p=0.790). Demographic and clinicopathological characteristics were not associated with NEAT1 expression in rectal cancer tissue. Conclusion: The obtained results suggest that the long noncoding RNA NEAT1 cannot be considered as a biomarker with diagnostic potential or for predicting response to nCRT in patients with rectal cancer. Validation of the current results in a larger group of patients with locally advanced rectal cancer is warranted

SMAD7 and SMAD4 expression in colorectal cancer progression and therapy response

Inhibitory SMAD7 and common mediator SMAD4 play crucial roles in SMAD-dependent TGF-I3 signaling that is often disrupted in colorectal cancer (CRC). This study aimed to profile the expression of SMAD7 and SMAD4 in primary and metastatic CRC and to evaluate their significance in disease progression and therapy response. The expression of SMAD7 and SMAD4 genes was analyzed by quantitative real-time PCR in tissues from 35 primary and metastatic CRC patients and in vitro in 7 human cell lines originating from colon tissue. Expression levels of SMAD7 and SMAD4, as well as their ratio, were determined and their association with tumor characteristics and response to therapy were evaluated. SMAD4 level was significantly lower in tumors compared to non-tumor tissues in both primary (p = 0.001) and metastatic (p = 0.001) CRC patients, while tumor expression of SMAD7 was significantly lower from non-tumor tissue only in metastatic patients (p = 0.017). SMAD7/SMAD4 ratio was elevated in CRC primary tumor tissues and cell lines compared to corresponding non-tumor tissues and cell line, respectively (p = 0.003). SMAD7 expression was significantly elevated in primary tumor tissues obtained from responders to neoadjuvant chemoradiotherapy (nCRT) compared to non-responders (p = 0.014). Alterations of expression and ratio of SMAD7 and SMAD4 in CRC cell lines, primary rectal cancer, and liver metastasis emphasize the importance of these genes in different stages of disease progression. Differential expression of SMAD7 in responders versus non-responders to nCRT should be further investigated for its potential predictive value