Molecular and Cellular Probes最新文献

{"title":"DNA in fresh urine supernatant is not affected by additional centrifugation and is protected against deoxyribonuclease","authors":"Ľubica Janovičová ,&nbsp;Katarína Kmeťová ,&nbsp;Ľubomíra Tóthová ,&nbsp;Barbora Vlková ,&nbsp;Peter Celec","doi":"10.1016/j.mcp.2023.101900","DOIUrl":"10.1016/j.mcp.2023.101900","url":null,"abstract":"<div><p>Urinary DNA is widely studied as a non-invasive marker for monitoring of kidneys after transplantation or the progression of urinary tract tumors. The quantity of urinary DNA especially of mitochondrial origin has been reported to mirror kidney damage in various renal diseases and their models. Processing of samples might affect urinary DNA concentrations but the details are not clear.</p><p>Samples of urine were collected from fifteen healthy volunteers. DNA was extracted from the whole urine, but also from the supernatant after centrifugation at 1600 <em>g</em> and 16000 g. In addition, we have analyzed the DNA in the microparticles in the pellet after the last spin. DNA was measured using fluorometry and real time PCR targeting nuclear and mitochondrial sequences. Addition of deoxyribonuclease to aliquots of samples enabled the characterization of DNA protection.</p><p>Centrifugation at 1600 <em>g</em> decreased the concentration of extracted DNA by 66% at least in samples with higher DNA in whole urine. Interestingly, the additional spin at 16000 g did not result in a significant decrease in DNA concentration in the supernatant despite detectable microparticle-associated DNA. Deoxyribonuclease decreases total and nuclear DNA by 26% and 31% in whole urine. The majority of urinary mitochondrial DNA seems to be protected against deoxyribonuclease.</p><p>Our results indicate high variability in urinary DNA even in healthy probands. Extracellular urinary DNA is partially bound to cell debris or microparticles, but a considerable part is still in the supernatant and is protected against cleavage. Further research should identify the nature of the protection, especially for mitochondrial DNA. Better understanding of the biology of urinary DNA should help its clinical interpretation.</p></div>","PeriodicalId":49799,"journal":{"name":"Molecular and Cellular Probes","volume":"68 ","pages":"Article 101900"},"PeriodicalIF":3.3,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9292307","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Potential of metagenomic next-generation sequencing in detecting infections of ICU patients","authors":"Yanxu Liang,&nbsp;Qingguo Feng,&nbsp;Kai Wei,&nbsp;Xiaoming Hou,&nbsp;Xiaotao Song,&nbsp;Yuantao Li","doi":"10.1016/j.mcp.2023.101898","DOIUrl":"10.1016/j.mcp.2023.101898","url":null,"abstract":"<div><h3>Background</h3><p>Due to the limitations of traditional microbiological detection techniques in evaluating complicated infections in ICU patients, it is necessary to explore novel and effective methods to improve the clinical detection of ICU patients’ infections.</p></div><div><h3>Objective</h3><p>This study aimed to evaluate the efficiency and specificity of mNGS in screening pathogens in the blood, deep phlegm, urine, and other sample types of ICU patients exploring an effective method for infection detection.</p></div><div><h3>Methods</h3><p>A total of 56 ICU patients with 131 samples were included in this study. The sample types included blood, deep phlegm, urine, drainage, anal swabs, and other types. Samples were analyzed by both conventional detection method and mNGS tests. The diagnosis efficiency and consistency of the two methods were compared. The distribution of the identified pathogens was analyzed. Moreover, the clinical features of patients with mNGS-positive or mNGS-negative results were compared.</p></div><div><h3>Results</h3><p>The positive rate of mNGS was 81.7% (107/131) including 3.1% (4/131) weakly positive, while the positive rate of traditional detection was only 30.5%, including 29 strong positive results and 11 weak positive results. Additionally, there were 41 patients chose to adjust anti-infection strategies according to the results of mNGS, which significantly saved treatment costs. The mNGS-positive patients showed a shorter ICU hospitalization and higher intention to adjust anti-infection strategies than the mNGS-negative patients.</p></div><div><h3>Conclusion</h3><p>mNGS is of great potential for the pathogen detection of ICU patients, and has a higher detection rate than traditional detection methods. Further clinical application investigations can be carried out to expand the application of mNGS.</p></div>","PeriodicalId":49799,"journal":{"name":"Molecular and Cellular Probes","volume":"68 ","pages":"Article 101898"},"PeriodicalIF":3.3,"publicationDate":"2023-04-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9344821","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Metagenomic next-generation sequencing for detection of pathogens in children with hematological diseases complicated with infection","authors":"Yating Zhang ,&nbsp;Dunhua Zhou ,&nbsp;Han Xia ,&nbsp;Jian Wang ,&nbsp;Huaqing Yang ,&nbsp;Luhong Xu ,&nbsp;Ke Huang ,&nbsp;Jianpei Fang","doi":"10.1016/j.mcp.2022.101889","DOIUrl":"10.1016/j.mcp.2022.101889","url":null,"abstract":"<div><h3>Objective</h3><p>Infection is one of the most common causes of death in children with hematological diseases. Here, we aim to investigate the value of metagenomic next-generation sequencing (mNGS) in the detection of causative pathogens in children with hematological diseases.</p></div><div><h3>Methods</h3><p>In this retrospective study, specimens from children with hematological diseases, who were admitted to Sun Yat-Sen University between June 2019 and September 2021, were collected for culture and mNGS.</p></div><div><h3>Results</h3><p>A total of 67 pediatric patients were enrolled, and 96 specimens were collected. The positive rate of mNGS was significantly higher than that of culture (57.2% vs 12.5%, <em>P</em> &lt; 0.01). The concordance (90.9%, 10/11) between the positive results of the two methods was high. mNGS detected more cases with <em>Pneumocystis jeroveci</em>, <em>Aspergillus flavus</em>, viruses, and some rare pathogens than culture. Mixed infections were detected by mNGS in 16 cases. Clinical anti-infective treatment was adjusted according to the results of mNGS, the conditions of most patients improved.</p></div><div><h3>Conclusion</h3><p>Compared to culture, mNGS shows great advantages in diagnosing bacterial, fungal, viral, and mixed infections in children with hematologic diseases, positively impacting clinical care. mNGS can be used as a complement to culture for pathogen detection.</p></div>","PeriodicalId":49799,"journal":{"name":"Molecular and Cellular Probes","volume":"67 ","pages":"Article 101889"},"PeriodicalIF":3.3,"publicationDate":"2023-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9498411","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Diagnostic and prognostic value of plasma cell-free DNA combined with VEGF-C in laryngeal squamous cell carcinoma","authors":"Qiang Huang ,&nbsp;Mengyou Ji ,&nbsp;Feiran Li ,&nbsp;Yufeng Li ,&nbsp;Xuehua Zhou ,&nbsp;Chi-yao Hsueh ,&nbsp;Liang Zhou","doi":"10.1016/j.mcp.2023.101895","DOIUrl":"10.1016/j.mcp.2023.101895","url":null,"abstract":"<div><h3>Background</h3><p>Circulating cell-free DNA (cfDNA) and vascular endothelial growth factor-C (VEGF-C) can be utilized to detect cancer and predict its prognosis. However, their potential application in laryngeal squamous cell carcinoma (LSCC) is unclear.</p></div><div><h3>Purpose</h3><p>This study aimed to identify the diagnostic and prognostic value of cfDNA and VEGF-C in LSCC patients<strong>.</strong></p></div><div><h3>Methods</h3><p>The plasma cfDNA of 148 LSCC patients and 43 non-tumor patients were isolated. Quantitative real-time PCR (qRT-PCR) was performed to assess long and short DNA fragments in plasma by amplifying the ALU repeats. ALU-qPCR results (ALU247/ALU115) were used to calculate cfDNA integrity index. Vascular endothelial growth factor-C (VEGF-C) level was detected by ELISA assay. Correlation between cfDNA and clinical features was analyzed. For detecting the sensitivity and specificity of cfDNA and VEGF-C alone or in combination for diagnosing LSCC, receiver operator characteristic (ROC) was established. For evaluating the overall survival (OS) of LSCC, Kaplan-Meier curves were established.</p></div><div><h3>Results</h3><p>LSCC patients had significantly higher levels of plasma cfDNA (ALU115, ALU247, and cfDNA integrity index) and VEGF-C than those without cancer (p &lt; 0.05), showing area under the curve (AUC) values of 0.79, 0.74, 0.62 and 0.80, when cutoff value was correspondingly defined at 2.14 ng/mL, 1.39 ng/mL, 0.73 and 412.90 pg/mL, respectively. The AUC for distinguishing LSCC patients from non-tumor patients by plasma cfDNA combined with VEGF-C was 0.89 (95% CI: 0.83–0.94). A significant correlation was found between plasma cfDNA levels and Ki-67, tumor size, pT stage, and smoking history (p &lt; 0.05). Based on survival analysis, low VEGF-C concentration groups had longer OS than those with high VEGF-C concentration (p = 0.02).</p></div><div><h3>Conclusion</h3><p>Indicators such as plasma cfDNA and VEGF-C may be used to diagnose and monitor LSCC for its noninvasiveness and rapid accessibility.</p></div>","PeriodicalId":49799,"journal":{"name":"Molecular and Cellular Probes","volume":"67 ","pages":"Article 101895"},"PeriodicalIF":3.3,"publicationDate":"2023-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9129727","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Liquid biopsy in the clinical management of cancer patients","authors":"Orsolya Biró","doi":"10.1016/j.mcp.2023.101892","DOIUrl":"10.1016/j.mcp.2023.101892","url":null,"abstract":"","PeriodicalId":49799,"journal":{"name":"Molecular and Cellular Probes","volume":"67 ","pages":"Article 101892"},"PeriodicalIF":3.3,"publicationDate":"2023-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9498435","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"MiR-128-3p as blood based liquid biopsy biomarker in childhood acute lymphoblastic leukemia","authors":"Andrea Rzepiel ,&nbsp;Anna Horváth ,&nbsp;Nóra Kutszegi ,&nbsp;András Gézsi ,&nbsp;Judit C. Sági ,&nbsp;Laura Almási ,&nbsp;Bálint Egyed ,&nbsp;Péter Lőrincz ,&nbsp;Tamás Visnovitz ,&nbsp;Gábor T. Kovács ,&nbsp;Csaba Szalai ,&nbsp;Ágnes F. Semsei ,&nbsp;Dániel J. Erdélyi","doi":"10.1016/j.mcp.2023.101893","DOIUrl":"10.1016/j.mcp.2023.101893","url":null,"abstract":"<div><h3>Background</h3><p>Minimal residual disease (MRD) is one of the most valuable independent prognostic factors in acute lymphoblastic leukemia (ALL). Bone marrow (BM) aspiration, however, is an invasive process. Previous studies have shown that microRNAs (miR) and extracellular vesicle (EV)-related miRs show different expression profiles at the presence of malignant cells compared to healthy controls. In our previous project, we have reported that two miRs previously described to be overexpressed in blasts were significantly decreased over the first week of the therapy of patients with ALL in the platelet free plasma fraction (PFP) of peripheral blood samples (PB). The aim of the current study was to assess the relation between day 15 flow cytometry (FC) MRD and expression of miR-128-3p and miR-222-3p miRs in exosome-enriched fraction (EEF) of PFP to evaluate whether their expression in EEF correlates with day 15 FC MRD more precisely.</p></div><div><h3>Methods</h3><p>PB was collected from 13 patients diagnosed with pediatric pre-B ALL at 4 time points. Expression of miR-128-3p and miR-222-3p was measured by qPCR in PFP and EEF.</p></div><div><h3>Results</h3><p>Positive correlation was found between changes of miR-128-3p expression in EEF or PFP by day 8 of chemotherapy and day 15 FC MRD (r_EEF = 0.99, p_EEF = 1.13E-9 and r_PFP = 0.99, p_PFP = 4.75E-9, respectively). Furthermore, the decrease of miR-128-3p in EEF by day 15 of treatment also showed a positive correlation with day 15 FC MRD (r_EEF = 0.96; p_EEF = 4.89E-5).</p></div><div><h3>Conclusion</h3><p>Our results show that circulating miRs are potential biomarkers of ALL MRD, asmiR-128-3p level both in PFP and EEF predicts day 15 FC MRD. In addition, the assessment of the EEF gave a more promising result.</p></div>","PeriodicalId":49799,"journal":{"name":"Molecular and Cellular Probes","volume":"67 ","pages":"Article 101893"},"PeriodicalIF":3.3,"publicationDate":"2023-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9128201","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Development of a specific and sensitive diagnostic PCR for rapid molecular authentication of the medicinal plant Portulaca oleracea","authors":"Mo-Rong Xu ,&nbsp;Meng-Shiou Lee ,&nbsp;Bo-Cheng Yang ,&nbsp;Hsiu-Chi Chang ,&nbsp;Chao-Lin Kuo ,&nbsp;Chia-Hsin Lin ,&nbsp;Hsi-Jien Chen ,&nbsp;Jai-Hong Cheng ,&nbsp;Fang-Chun Sun","doi":"10.1016/j.mcp.2022.101890","DOIUrl":"10.1016/j.mcp.2022.101890","url":null,"abstract":"<div><p>Adulteration by <em>Bacopa monnieri</em> (BM) in <em>Portulaca oleracea</em> (PO) plants frequently occurs; it decreases the efficacy of traditional Chinese medicine (TCM) and leads to fraud in the herbal marketplace. In this study, a diagnostic PCR assay was established for the rapid authentication of PO and BM in the herbal market. The sequence divergences in internal transcribed spacer 2 (ITS2) between PO and its adulterant species were used to design diagnostic PCR primers. The specific designed primer sets were evaluated and show that the diagnostic PCR assay can be used to verify the authenticity of PO and BM. The detection limits of the primer set for PO and BM identification were 10 pg and 1 pg, respectively. The reactivity of diagnostic PCR was 0.1% PO genomic DNA and 0.01% BM genomic DNA in the test sample during DNA amplification. In addition, multiplex PCR (mPCR) for PO and BM identification was also established. The samples were more susceptible to the effect of steaming in authentication by singleplex PCR and mPCR than boiling and drying treatment. Furthermore, commercial samples from the market were used to demonstrate the applicability of the developed diagnostic PCR for PO authentication and diagnose BM adulteration, and the investigation found that approximately 72.2% (13/18) of PO plants in the herbal market were adulterated. In conclusion, the diagnostic PCR assay was successfully developed and its specificity, sensitivity and reactivity for PO and BM authentication were proven. These developed PCR-based molecular methods can be applied as an identification tool for PO authenticity and can be practically applied for inspection of BM adulteration in the herbal market in the future.</p></div>","PeriodicalId":49799,"journal":{"name":"Molecular and Cellular Probes","volume":"67 ","pages":"Article 101890"},"PeriodicalIF":3.3,"publicationDate":"2023-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9129310","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"TYMSOS-miR-101-3p-NETO2 axis promotes osteosarcoma progression","authors":"Zun Zhang ,&nbsp;Jin Wang ,&nbsp;Xiaoyan Zhang ,&nbsp;Bo Ran ,&nbsp;Jie Wen ,&nbsp;Hong Zhang","doi":"10.1016/j.mcp.2022.101887","DOIUrl":"10.1016/j.mcp.2022.101887","url":null,"abstract":"<div><h3>Background</h3><p>Osteosarcoma (OS) is a type of bone cancer most often affects pre-teens and teens, but it is still a rare disorder. Neuropilin and tolloid-like 2 (NETO2) has been reported to promote OS progression, but its upstream mechanism in OS cells remains obscure.</p></div><div><h3>Methods</h3><p>Quantitative real-time PCR (RT-qPCR) and Western blot were conducted to examine RNA and protein levels, separately. Functional assays were performed to assess the impact of NETO2 on OS cell malignancy. Moreover, bioinformatics analyses and mechanism experiments were performed to identify the upstream mechanism of NETO2 in OS cells.</p></div><div><h3>Results</h3><p>Functionally, NETO2 depletion repressed cell proliferation, migration and invasion as well as epithelial-mesenchymal transition (EMT) but triggered the apoptosis of OS cells. NETO2 is directly targeted and negatively regulated by microRNA-101-3p (miR-101-3p). Mechanically, miR-101-3p could combine with long noncoding RNA (lncRNA) TYMS opposite strand RNA (TYMSOS) in OS cells. In addition, our study proved that TYMSOS promotes the malignancy of OS via elevating NETO2 expression as miR-101-3p sponge.</p></div><div><h3>Conclusion</h3><p>TYMSOS-miR-101-3p-NETO2 axis promotes the malignant behaviors of OS cells, which might offer a novel sight for OS treatment.</p></div>","PeriodicalId":49799,"journal":{"name":"Molecular and Cellular Probes","volume":"67 ","pages":"Article 101887"},"PeriodicalIF":3.3,"publicationDate":"2023-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9127744","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Identification of liquid biopsy-based mutations in colorectal cancer by targeted sequencing assays","authors":"István Szász ,&nbsp;Tímea Kiss ,&nbsp;Attila Mokánszki ,&nbsp;Viktória Koroknai ,&nbsp;János Deák ,&nbsp;Vikas Patel ,&nbsp;Krisztina Jámbor ,&nbsp;Róza Ádány ,&nbsp;Margit Balázs","doi":"10.1016/j.mcp.2022.101888","DOIUrl":"10.1016/j.mcp.2022.101888","url":null,"abstract":"<div><p>Recently, liquid biopsy, as a promising approach was introduced for the analysis of different tumor-derived circulating markers including tumor DNA and cell free DNA (ct/cfDNA). Identification of mutations in cfDNA may allow the early detection of tumors, as well as predicting and monitoring treatment responses in a minimally invasive way. In the present study, we used commercially available gene panels to verify the mutation overlap between liquid biopsy and abnormalities detected in colorectal tumor tissue. The two panels (Archer®VariantPlex®Solid Tumor and LIQUIDPlexTM ctDNA) overlap in 23 genes, which enables a comprehensive view of tumor-plasma mutational status by next generation sequencing. We successfully analyzed 16 plasma and 16 tumor samples. We found that 87% of tumor tissues contained 44 mutations in 12 genes and 43.8% of cfDNA harbored 13 mutations in 5 genes. To verify whether the mutation pattern of the tumor DNA could be consistently detected in plasma cfDNA, we compared the alterations between cfDNA and matched tissue DNA in nine patients. Six of the 9 tumor tissues harbored mutations in <em>TP53, KRAS</em> or <em>MET</em> genes, those were not detectable by the ctDNA kit, even eventhough the exons of these genes overlap in both panels. Comparing the mutational patterns of the matched samples, we found that only one cfDNA had the same mutations (<em>KRAS, SMAD4</em> and <em>TP53</em>) in the paired tissue. The results of the comparison between tumor tissue DNA and matched plasma cfDNA underline the importance of studying the paired solid tumor and plasma samples together.</p></div>","PeriodicalId":49799,"journal":{"name":"Molecular and Cellular Probes","volume":"67 ","pages":"Article 101888"},"PeriodicalIF":3.3,"publicationDate":"2023-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9498409","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}

{"title":"Extracellular vesicles derived from dental mesenchymal stem/stromal cells with gemcitabine as a cargo have an inhibitory effect on the growth of pancreatic carcinoma cell lines in vitro","authors":"Daniela Klimova ,&nbsp;Jana Jakubechova ,&nbsp;Ursula Altanerova ,&nbsp;Andreas Nicodemou ,&nbsp;Jakub Styk ,&nbsp;Tomas Szemes ,&nbsp;Vanda Repiska ,&nbsp;Cestmir Altaner","doi":"10.1016/j.mcp.2023.101894","DOIUrl":"10.1016/j.mcp.2023.101894","url":null,"abstract":"<div><p>Extracellular vesicles (EVs) are nowadays a target of interest in cancer therapy as a successful drug delivering tool. Based on their many beneficial biocompatible properties are designed to transport nucleic acids, proteins, various nanomaterials or chemotherapeutics. Extracellular vesicles derived from mesenchymal stem/stromal cells (MSCs) possess their tumor-homing abilities. This inspired us to engineer the MSC's EVs to be packed with chemotherapeutic agents and deliver it as a Trojan horse directly into tumor cells. In our study, human dental pulp MSCs (DP-MSCs) were cultivated with gemcitabine (GCB), which led to its absorption by the cells and subsequent secretion of the drug out into conditioned media in EVs. Concentrated conditioned media containing small EVs (potentially exosomes) significantly inhibited the cell growth of pancreatic carcinoma cell lines <em>in vitro</em>. DP-MSCs were simultaneously engineered to express a suicide gene fused yeast cytosinedeaminase:uracilphosphoribosyltransferase (<em>yCD::UPRT).</em> The product of the suicide gene converts non-toxic prodrug 5-fluorocytosine (5-FC) to highly cytotoxic chemotherapeutic drug 5-fluorouracil (5-FU) in the recipient cancer cells. Conversion of 5-FC to 5-FU had an additional effect on cancer cell's growth inhibition. Our results showed a therapeutic potential for DP-MSC-EVs to be designed for successful delivering of chemotherapeutic drugs, together with prodrug suicide gene therapy system.</p></div>","PeriodicalId":49799,"journal":{"name":"Molecular and Cellular Probes","volume":"67 ","pages":"Article 101894"},"PeriodicalIF":3.3,"publicationDate":"2023-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"9483128","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":3,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}