OBJECTIVES: Small extracellular vesicles (EVs) contain various signaling molecules, thus playing a crucial role in cell-to-cell communication and emerging as a promising source of biomarkers. However, the lack of standardized procedures impedes their translation to clinical practice. Thus, we compared different approaches for high-throughput analysis of small EVs transcriptome. METHODS: Small EVs were isolated from 150 μL of serum. Quality and quantity were assessed by dynamic light scattering, transmission electron microscopy, and Western blot. Comparison of RNA extraction efficiency was performed, and expression of selected genes was analyzed by RT-qPCR. Whole transcriptome analysis was done using microarrays. RESULTS: Obtained data confirmed the suitability of size exclusion chromatography for isolation of small EVs. Analyses of gene expression showed the best results in case of samples isolated by Monarch Total RNA Miniprep Kit. Totally, 7,182 transcripts were identified to be deregulated between colorectal cancer patients and healthy controls. The majority of them were non-coding RNAs with more than 70 % being lncRNAs, while protein-coding genes represented the second most common gene biotype. CONCLUSIONS: We have optimized the protocol for isolation of small EVs and their RNA from low volume of sera and confirmed the suitability of Clariom D Pico Assays for transcriptome profiling.
- MeSH
- extracelulární vezikuly * genetika metabolismus MeSH
- gelová chromatografie MeSH
- lidé MeSH
- RNA MeSH
- stanovení celkové genové exprese * metody MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Colorectal cancer (CRC) is the second most prevalent cancer type worldwide, which highlights the urgent need for non-invasive biomarkers for its early detection and improved prognosis. We aimed to investigate the patterns of long non-coding RNAs (lncRNAs) in small extracellular vesicles (sEVs) collected from low-volume blood serum specimens of CRC patients, focusing on their potential as diagnostic biomarkers. Our research comprised two phases: an initial exploratory phase involving RNA sequencing of sEVs from 76 CRC patients and 29 healthy controls, and a subsequent validation phase with a larger cohort of 159 CRC patients and 138 healthy controls. Techniques such as dynamic light scattering, transmission electron microscopy, and Western blotting were utilized for sEV characterization. Optimized protocol for sEV purification, RNA isolation and preamplification was applied to successfully sequence the RNA content of sEVs and validate the results by RT-qPCR. We successfully isolated sEVs from blood serum and prepared sequencing libraries from a low amount of RNA. High-throughput sequencing identified differential levels of 460 transcripts between CRC patients and healthy controls, including mRNAs, lncRNAs, and pseudogenes, with approximately 20% being lncRNAs, highlighting several tumor-specific lncRNAs that have not been associated with CRC development and progression. The validation phase confirmed the upregulation of three lncRNAs (NALT1, AL096828, and LINC01637) in blood serum of CRC patients. This study not only identified lncRNA profiles in a population of sEVs from low-volume blood serum specimens of CRC patients but also highlights the value of innovative techniques in biomolecular research, particularly for the detection and analysis of low-abundance biomolecules in clinical samples. The identification of specific lncRNAs associated with CRC provides a foundation for future research into their functional roles in cancer development and potential clinical applications.
Calcium channel blockers are among the most commonly used agents in the treatment of cardiovascular diseases. There are several known side-effects associated with their long-term use, whereas other potential adverse effects are yet to be proven. This study aims to evaluate the association between calcium channel blockers exposure and the incidence of second primary malignancy. We established a cohort of 1401 patients with colorectal cancer diagnosed in our institution between January 2003 and December 2016. Patients were followed-up until December 2020. The tumor characteristics and basic clinical data including medication information were obtained from the hospital information system database. Second malignancy was detected in 301 patients (21.5%), and occurred in 27.8% of patients who used calcium channel blockers compared to only 19.9% among non-users. Their use was associated with an increased incidence of bladder cancer in particular. Subanalysis of patients with second malignancy displayed a higher proportion of right-sided colon cancer compared to rectal carcinoma in non-users. Survival analysis revealed significantly better outcomes in early-stage colorectal cancer patients without a history of calcium channel blockers treatment or second primary malignancy.
- MeSH
- blokátory kalciových kanálů MeSH
- kardiovaskulární nemoci * MeSH
- kolon MeSH
- lidé MeSH
- nádory rekta * MeSH
- nežádoucí účinky léčiv * MeSH
- sekundární malignity * MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
An increasing number of studies has brought evidence of the protective role of statin use against different types of cancer. However, data on their association with second primary malignancies (SPMs) are lacking. The purpose of this study was to determine the role of hypolipidemic treatment in the prevention of second primary cancer in colorectal cancer (CRC) survivors. We conducted a retrospective single-institution study of 1401 patients with newly diagnosed colorectal cancer from January 2003 to December 2016, with follow-up until December 2020. An SPM was detected in 301 patients (21%), and the incidence was significantly lower in patients with statin medication. However, stratification by cancer types revealed an increased incidence of bladder and gastric cancer in hypolipidemic users. A Kaplan-Meier analysis of early-stage CRC survivors with an SPM showed a significant survival benefit in patients without a history of hypolipidemic treatment. Despite the protective role of statins on overall second cancer incidence, these data indicate that CRC survivors treated with hypolipidemic drugs should be screened more cautiously for SPMs, especially for gastric and bladder cancer.
- Publikační typ
- časopisecké články MeSH
Úvod: Poškození kůže s hlubšími podkožními strukturami jsou častými nežádoucími účinky radioterapie. Limitují pak možnosti jejího pokračování a léčba následných komplikací je rutinní náplní praxe radiačního onkologa. Většinově se jedná o projevy akutní s perspektivou zhojení v řádech týdnů ad integrum. Méně často, avšak nezřídka s vážnými projevy, se objevuje toxicita chronická v horizontu měsíců, nebo dokonce mnoha let po ozařování, mající podobu postradiačního vředu s potenciálem vzniku sekundární malignity. Stran chirurgie se může jednat o jednu z nejobtížněji léčitelných chronických lézí, kdy je v extrémních případech k jejich vyléčení potřebná extenzivní resekce celé oblasti defektu a hlubších struktur s navazujícím komplexním, plasticko-chirurgickým, výkonem. Kazuistika: Tato kazuistika prezentuje rychle progredující zhoršení chronických postradiačních změn v oblasti levého prsu, k němuž došlo po 33 let od radioterapie, a to po zcela nezávažné traumatizaci kůže. Před tímto zvratem byly u pacientky v rámci dispenzarizace popsány pouze dlouhodobě stacionární změny běžného charakteru. Po vleklém ambulantním ošetřování defektu s neuspokojivými výsledky, kdy pacientka opakovaně odmítala radikální výkon ve smyslu sanační mastektomie, nakonec stav pro rychlou progresi nálezu vyústil v komplexní chirurgický výkon s nutnou spoluprací mammárního, plastického a hrudního chirurga. Závěr: Ačkoli se změny podobného charakteru vzácněji vyskytují i s odstupem mnoha let s podobnou závažností, nenacházíme v literatuře popsán podobně dramatický vývoj a následnou progresi po třech desetiletích od terapie, vyžadující takto rozsáhlou intervenci.
Introduction: Damage of the skin and its underlying structures is a common side effect of radiotherapy. These conditions limit further treatment and dealing with these complications is a routine practice of clinical oncologist. The majority of the complications are immediate, with a perspective of healing ad integrum within a few weeks. Less frequently, but sometimes with severe manifestations, chronic toxicity occurs belatedly after months, or even many years after irradiation, in form of post-radiation ulcer, for instance with potential of secondary malignant transformation. Regarding surgery, it might be one of the most challenging chronic wounds to treat. In extreme cases, extensive resection of the entire affected area is needed, inevitably ending with demanding reconstruction of the resulting defect. Case report: This case report presents a female patient with rapidly progressive post-radiation chest wall defect 33 years after the irradiation, when relatively insignificant skin injury occured. Prior to this sudden deterioration, only long-term, non-progressive changes, without a cutaneous defect, were described during the dispensarisation. After a protracted outpatient treatment with unsatisfactory results, when the patient repeatedly refused mastectomy, the condition inevitably led to the complex surgical procedure with necessary cooperation of breast, plastic and thoracic surgeons. Conclusion: Although changes of the similar severity rarely occur even after many years following the treatment, we haven‘t found such a dramatic change of the patient’s condition three decades after the therapy with urgency of this type of complex, surgical intervention in current literature.
- Klíčová slova
- postradiační defekt,
- MeSH
- lidé MeSH
- mastektomie rozsáhlá radikální metody MeSH
- nádory prsu * chirurgie radioterapie MeSH
- plastická chirurgie metody MeSH
- radiační poranění chirurgie etiologie MeSH
- radioterapie * škodlivé účinky MeSH
- senioři nad 80 let MeSH
- výsledek terapie MeSH
- Check Tag
- lidé MeSH
- senioři nad 80 let MeSH
- ženské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
- práce podpořená grantem MeSH
