The 3-mercaptopyruvate sulfurtransferase (MPST) is a protein persulfidase, occurring mainly in mitochondria. Although function of this protein in cancer cells has been already studied, no clear outcome can be postulated up to now. Therefore, we focused on the determination of function of MPST in colon (HCT116 cells)/colorectal (DLD1 cells) cancers. In silico analysis revealed that in gastrointestinal cancers, MPST together with its binding partners can be either of a high risk or might have a protective effect. Silencing of MPST gene resulted in decreased ATP, while acetyl-CoA levels were elevated. Increased apoptosis was detected in cells with silenced MPST gene, which was accompanied by decrease in mitochondrial membrane potential, but no changes in IP3 receptor's protein. Mitochondria underwent activation of fission and elevated DRP1 expression after MPST silencing. Proliferation and migration of DLD1 and HCT116 cells were markedly affected, showing the importance of MPST protein in colon/colorectal cancer development.
- Klíčová slova
- 3- mercaptopyruvate sulfurtransferase, Colon/colorectal cancer, Fission, Mitochondria,
- MeSH
- apoptóza MeSH
- dynaminy metabolismus genetika MeSH
- HCT116 buňky MeSH
- kolorektální nádory * metabolismus patologie enzymologie genetika MeSH
- lidé MeSH
- membránový potenciál mitochondrií MeSH
- mitochondrie metabolismus MeSH
- nádorové buněčné linie MeSH
- nádory tračníku metabolismus patologie genetika enzymologie MeSH
- pohyb buněk MeSH
- proliferace buněk MeSH
- sulfurtransferasy * metabolismus genetika MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- 3-mercaptopyruvate sulphurtransferase MeSH Prohlížeč
- DNM1L protein, human MeSH Prohlížeč
- dynaminy MeSH
- sulfurtransferasy * MeSH
Paclitaxel (PTX) is a chemotherapeutic agent affecting microtubule polymerization. The efficacy of PTX depends on the type of tumor, and its improvement would be beneficial in patients' treatment. Therefore, we tested the effect of slow sulfide donor GYY4137 on paclitaxel sensitivity in two different breast cancer cell lines, MDA-MB-231, derived from a triple negative cell line, and JIMT1, which overexpresses HER2 and is resistant to trastuzumab. In JIMT1 and MDA-MB-231 cells, we compared IC50 and some metabolic (apoptosis induction, lactate/pyruvate conversion, production of reactive oxygen species, etc.), morphologic (changes in cytoskeleton), and functional (migration, angiogenesis) parameters for PTX and PTX/GYY4137, aiming to determine the mechanism of the sensitization of PTX. We observed improved sensitivity to paclitaxel in the presence of GYY4137 in both cell lines, but also some differences in apoptosis induction and pyruvate/lactate conversion between these cells. In MDA-MB-231 cells, GYY4137 increased apoptosis without affecting the IP3R1 protein, changing the morphology of the cytoskeleton. A mechanism of PTX sensitization by GYY4137 in JIMT1 cells is distinct from MDA-MB-231, and remains to be further elucidated. We suggest different mechanisms of action for H2S on the paclitaxel treatment of MDA-MB-231 and JIMT1 breast cancer cell lines.
- Klíčová slova
- apoptosis, breast cancer cell lines, metabolism, paclitaxel, slow sulfide donor,
- MeSH
- apoptóza * účinky léků MeSH
- chemorezistence účinky léků MeSH
- lidé MeSH
- morfoliny * farmakologie MeSH
- nádorové buněčné linie MeSH
- nádory prsu * farmakoterapie patologie metabolismus MeSH
- organothiofosforové sloučeniny * farmakologie MeSH
- paclitaxel * farmakologie MeSH
- reaktivní formy kyslíku metabolismus MeSH
- sulfidy farmakologie MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- GYY 4137 MeSH Prohlížeč
- morfoliny * MeSH
- organothiofosforové sloučeniny * MeSH
- paclitaxel * MeSH
- reaktivní formy kyslíku MeSH
- sulfidy MeSH
Chronic hyperplastic candidiasis (CHC) presents a distinctive and relatively rare form of oral candidal infection characterized by the presence of white or white-red patches on the oral mucosa. Often mistaken for leukoplakia or erythroleukoplakia due to their appearance, these lesions display nonhomogeneous textures featuring combinations of white and red hyperplastic or nodular surfaces. Predominant locations for such lesions include the tongue, retro-angular mucosa, and buccal mucosa. This paper aims to investigate the potential influence of specific anatomical locations, retro-angular mucosa, on the development and occurrence of CHC. By examining the relationship between risk factors, we present an approach based on machine learning (ML) to predict the location of CHC occurrence. In this way, we employ Gradient Boosting Regression (GBR) to classify CHC lesion locations based on important risk factors. This estimator can serve both research and diagnostic purposes effectively. The findings underscore that the proposed ML technique can be used to predict the occurrence of CHC in retro-angular mucosa compared to other locations. The results also show a high rate of accuracy in predicting lesion locations. Performance assessment relies on Mean Squared Error (MSE), Root Mean Squared Error (RMSE), R-squared (R2), and Mean Absolute Error (MAE), consistently revealing favorable results that underscore the robustness and dependability of our classification method. Our research contributes valuable insights to the field, enhancing diagnostic accuracy and informing treatment strategies.
BACKGROUND: Cystathionine β-synthase (CBS), one of three enzymes that endogenously produce hydrogen sulfide, is extensively studied for its relevance in the cells of various tumors. In our previous work, we observed that the immunofluorescence pattern of CBS is very similar to that of tubulin and actin. Therefore, we focused on the potential interaction of CBS with cytoskeletal proteins β-actin and β-tubulin and the functional relevance of the potential interaction of these proteins in colorectal carcinoma cell lines. METHODS: To study the potential interaction of CBS with cytoskeletal proteins and its functional consequences, a CBS-knockout DLD1 (DLDx) cell line was established by using the CRISPR/Cas9 gene editing method. The interaction of the selected cytoskeletal protein with CBS was studied by immunoprecipitation, Western blot analysis, immunofluorescence, and proximity ligation assay. The functional consequences were studied by proliferation and migration assays and by generation of xenografts in SCID/bg mice. RESULTS: We have found that CBS, an enzyme that endogenously produces H2S, binds to cytoskeletal β-tubulin and, to a lesser extent, also to β-actin in colorectal carcinoma-derived cells. When CBS was knocked out by the CRISPR/Cas9 technique (DLDx), we observed a de-arranged cytoskeleton compared to the unmodified DLD1 cell line. Treatment of these cells with a slow sulfide donor GYY4137 resulted in normal organization of the cytoskeleton, thus pointing to the role of CBS in microtubule dynamics. To evaluate the physiological importance of this observation, both DLD1 and DLDx cells were injected into SCID/bg mice, and the size and mass of the developed xenografts were evaluated. Significantly larger tumors developed from DLDx compared to the DLD1 cells, which correlated with the increased proliferation of these cells. CONCLUSIONS: Taken together, in colorectal cancer DLD1 cells, CBS binds to the cytoskeleton, modulates microtubule dynamics, and thus affects the proliferation and migration in the colorectal carcinoma stable cell line.
- Klíčová slova
- colorectal carcinoma cells, cystathionine beta-synthase, cytoskeleton, xenografts, β-tubulin,
- Publikační typ
- časopisecké články MeSH
BACKGROUND: Paraneoplastic pemphigus (PNP) in the oral cavity is a rare variant of blistering pemphigus disease closely associated with mostly malignant tumors. The diagnosis may even precede an underlying malignancy enabling early detection. Here, we describe a previously unreported case of PNP associated with HPV-related tonsillar squamous cell carcinoma. METHODS AND RESULTS: A 50-year-old woman was referred to a dentist because of painful oral lesions resembling aphthae major and minor. Later, blisters appeared and an incisional biopsy was performed. Histological examination revealed an unusual coexistence of subepithelial and intraepithelial blisters raising suspicion of paraneoplastic pemphigus. The patient underwent 18F-FDG PET/MRI, showing a metabolically active process in the left palatal tonsil. Diagnostic biopsy revealed HPV type 16 associated tonsillar squamous cell carcinoma. A left tonsillectomy with elective left-sided neck dissection was performed. The postoperative period was complicated by bilateral fluidothorax. Two weeks after radical tumor removal, the mucosal and skin lesions of PNP disappeared. The patient currently shows no evidence of recurrence either of malignancy or PNP eight months after the surgery. CONCLUSION: PNP is a rare autoimmune blistering disease characterized by polymorphous cutaneous and mucosal lesions associated with internal neoplasms including HPV associated squamous cell carcinoma of a tonsil. In order to identify an occult malignancy, a whole-body PET/CT or PET/MRI scan is recommended. Rarely, accurate patient management may depend on the dentist being familiar with this entity and on interdisciplinary cooperation involving dermatologist, radiologist, pathologist, and pneumologist. A strict patient follow-up is indicated.
- Klíčová slova
- 18F-FDG PET/MRI, oral mucosa lesions; paraneoplastic autoimmune multiorgan syndrome, paraneoplastic pemphigus, pulmonary complications, tonsillar carcinoma HPV associated,
- MeSH
- autoimunitní nemoci * komplikace MeSH
- infekce papilomavirem * komplikace diagnóza MeSH
- krční mandle patologie MeSH
- lidé středního věku MeSH
- lidé MeSH
- paraneoplastické syndromy * etiologie diagnóza patologie MeSH
- pemfigus * diagnóza etiologie MeSH
- PET/CT škodlivé účinky MeSH
- puchýř komplikace MeSH
- spinocelulární karcinom * komplikace diagnóza MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- kazuistiky MeSH
Although paclitaxel (PTX) is potent chemotherapeutic agent commonly used in variety of cancers, in colorectal carcinoma its usage is excluded because of low effectivity. Up to now, some experimental attempts were utilized to improve sensitivity of colorectal carcinoma to PTX. We used a slow sulfide donor GYY4137 to increase sensitivity of colorectal carcinoma cells to PTX. As a model of colorectal carcinoma, we utilized three different cell lines - HCT116, SW620 and DLD1. We compared IC50 for PTX and PTX/GYY4137, cell cycle, apoptosis, ATP levels and changes in intracellular pH. We observed significant decrease in IC50 levels in PTX/GYY4137 groups compared to PTX in all three cell lines. PTX arrested cell cycle in G2/M phase. Differences in S phase were observed in HCT116 and DLD1 cells treated with 20 nM PTX/GYY4137, but not in SW620 cell. GYY4137 increased early, but not late phase of apoptosis. This increase was not detected in non-cancer EAHy926 cells. Upregulation of IP3R1 suggested involvement of these receptors in PTX and/or GYY4137 induced apoptosis. We also observed partial ATP depletion and intracellular acidification in PTX treated groups. In PTX/GYY4137 groups of all three cell lines no ATP depletion was detectable and intracellular acidification was lower than in PTX treated groups. Slight differences in all measured parameters were determined among HCT116, SW620 and DLD1 cells, which is probably due to physiological variations in these cells. Taking together, sensitivity of PTX to colorectal carcinoma cell lines could be increased by slow sulfide donor GYY4137, probably through potentiation of apoptosis.
- Klíčová slova
- Colorectal carcinoma cells, H(2)S, Paclitaxel, Slow sulfide donor GYY4137,
- MeSH
- adenosintrifosfát farmakologie MeSH
- apoptóza MeSH
- kolorektální nádory * patologie MeSH
- lidé MeSH
- morfoliny MeSH
- nádorové buněčné linie MeSH
- organothiofosforové sloučeniny MeSH
- paclitaxel farmakologie MeSH
- sulfan * metabolismus MeSH
- sulfidy farmakologie terapeutické užití MeSH
- Check Tag
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
- Názvy látek
- adenosintrifosfát MeSH
- GYY 4137 MeSH Prohlížeč
- morfoliny MeSH
- organothiofosforové sloučeniny MeSH
- paclitaxel MeSH
- sulfan * MeSH
- sulfidy MeSH
The retromandibular transparotid approach enables the most direct access to the central and posterior part of the mandibular ramus including the condylar region. So far it has not been widely used for the management of benign pathology of the mandible. The purpose of this study was to evaluate the utilization rate of this approach in the nontrauma setting including the determination of suitable indications for this access. In total, 105 patients with 107 retromandibular transparotid approaches performed in the 6.5 years (from May 2014 to November 2020) were evaluated. Patients suffering from nontrauma pathology accounted for 4.7% of all cases. The recurrences of different types of odontogenic cysts and secondary chronic osteomyelitis were surgically managed via this approach with aesthetically acceptable resultant scar achievement and no identified postoperative complications. All lesions resolved and no recurrences occurred during the follow-up 32.0 ± 20.7 months (range 6 to 59 months, median = 26 months). The retromandibular transparotid approach may be considered for the enucleation of benign bone lesions in selected patients. Another type of surgery for the management of benign nontraumatic conditions in an accessible area without requirements for continuity resection and jaw reconstruction may be also suitable for using this approach.
The sodium/calcium exchanger (NCX) is a unique calcium transport system, generally transporting calcium ions out of the cell in exchange for sodium ions. Nevertheless, under special conditions this transporter can also work in a reverse mode, in which direction of the ion transport is inverted-calcium ions are transported inside the cell and sodium ions are transported out of the cell. To date, three isoforms of the NCX have been identified and characterized in humans. Majority of information about the NCX function comes from isoform 1 (NCX1). Although knowledge about NCX function has evolved rapidly in recent years, little is known about these transport systems in cancer cells. This review aims to summarize current knowledge about NCX functions in individual types of cancer cells.
- Klíčová slova
- apoptosis, calcium, cancer cells, sodium-calcium exchanger,
- MeSH
- invazivní růst nádoru MeSH
- iontový transport MeSH
- lidé MeSH
- nádory metabolismus MeSH
- pumpa pro výměnu sodíku a vápníku metabolismus MeSH
- sodík metabolismus MeSH
- vápník metabolismus MeSH
- zvířata MeSH
- Check Tag
- lidé MeSH
- zvířata MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- přehledy MeSH
- Názvy látek
- pumpa pro výměnu sodíku a vápníku MeSH
- sodík MeSH
- vápník MeSH
Haloalkane dehalogenases can cleave a carbon-halogen bond in a broad range of halogenated aliphatic compounds. However, a highly conserved catalytic pentad composed of a nucleophile, a catalytic base, a catalytic acid, and two halide-stabilizing residues is required for their catalytic activity. Only a few family members, e.g., DsaA, DmxA, or DmrB, remain catalytically active while employing a single halide-stabilizing residue. Here, we describe a novel haloalkane dehalogenase, DsvA, from a mildly thermophilic bacterium, Saccharomonospora viridis strain DSM 43017, possessing one canonical halide-stabilizing tryptophan (W125). At the position of the second halide-stabilizing residue, DsvA contains the phenylalanine F165, which cannot stabilize the halogen anion released during the enzymatic reaction by a hydrogen bond. Based on the sequence and structural alignments, we identified a putative second halide-stabilizing tryptophan (W162) located on the same α-helix as F165, but on the opposite side of the active site. The potential involvement of this residue in DsvA catalysis was investigated by the construction and biochemical characterization of the three variants, DsvA01 (F165W), DsvA02 (W162F), and DsvA03 (W162F and F165W). Interestingly, DsvA exhibits a preference for the (S)- over the (R)-enantiomers of β-bromoalkanes, which has not been reported before for any characterized haloalkane dehalogenase. Moreover, DsvA shows remarkable operational stability at elevated temperatures. The present study illustrates that protein sequences possessing an unconventional composition of catalytic residues represent a valuable source of novel biocatalysts.IMPORTANCE The present study describes a novel haloalkane dehalogenase, DsvA, originating from a mildly thermophilic bacterium, Saccharomonospora viridis strain DSM 43017. We report its high thermostability, remarkable operational stability at high temperatures, and an (S)-enantiopreference, which makes this enzyme an attractive biocatalyst for practical applications. Sequence analysis revealed that DsvA possesses an unusual composition of halide-stabilizing tryptophan residues in its active site. We constructed and biochemically characterized two single point mutants and one double point mutant and identified the noncanonical halide-stabilizing residue. Our study underlines the importance of searching for noncanonical catalytic residues in protein sequences.
- Klíčová slova
- (S)-enantiopreference, catalytic residues, dehalogenase, enantioselectivity, halide-stabilizing residues, haloalkane, haloalkane dehalogenase, kinetics, mutagenesis, structure, substrate specificity, thermophilic bacterium, thermostability,
- MeSH
- Actinobacteria chemie genetika metabolismus MeSH
- bakteriální proteiny chemie genetika metabolismus MeSH
- hydrolasy chemie genetika metabolismus MeSH
- katalýza MeSH
- substrátová specifita MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- bakteriální proteiny MeSH
- haloalkane dehalogenase MeSH Prohlížeč
- hydrolasy MeSH
Several papers have reported that calcium channel blocking drugs were associated with increased breast cancer risk and worsened prognosis. One of the most common signs of breast tumors is the presence of small deposits of calcium, known as microcalcifications. Therefore, we studied the effect of dihydropyridine nifedipine on selected calcium transport systems in MDA-MB-231 cells, originating from triple negative breast tumor and JIMT1 cells that represent a model of HER2-positive breast cancer, which possesses amplification of HER2 receptor, but cells do not response to HER2 inhibition treatment with trastuzumab. Also, we compared the effect of nifedipine on colorectal DLD1 and ovarian A2780 cancer cells. Both, inositol 1,4,5-trisphosphate receptor type 1 (IP3R1) and type 1 sodium calcium exchanger (NCX1) were upregulated due to nifedipine in DLD1 and A2780 cells, but not in breast cancer MDA-MB-231 and JIMT1 cells. On contrary to MDA-MB-231 and JIMT1 cells, in DLD1 and A2780 cells nifedipine induced apoptosis in a concentration-dependent manner. After NCX1 silencing and subsequent treatment with nifedipine, proliferation was decreased in MDA-MB-231, increased in DLD1 cells, and not changed in JIMT1 cells. Silencing of IP3R1 revealed increase in proliferation in DLD1 and JIMT1 cells, but caused decrease in proliferation in MDA-MB-231 cell line after nifedipine treatment. Interestingly, after nifedipine treatment migration was not significantly affected in any of tested cell lines after NCX1 silencing. Due to IP3R1 silencing, significant decrease in migration occurred in MDA-MB-231 cells after nifedipine treatment, but not in other tested cells. These results support different function of the NCX1 and IP3R1 in the invasiveness of various cancer cells due to nifedipine treatment.
- Klíčová slova
- Apoptosis, Breast cancer, Inositol 1,4,5-trisphosphate receptor, Migration, Sodium calcium exchanger 1,
- MeSH
- apoptóza účinky léků genetika MeSH
- blokátory kalciových kanálů farmakologie MeSH
- inositol-1,4,5-trisfosfát - receptory genetika metabolismus MeSH
- kolorektální nádory genetika metabolismus patologie MeSH
- lidé MeSH
- nádorové buněčné linie MeSH
- nádory prsu genetika metabolismus patologie MeSH
- nádory vaječníků genetika metabolismus patologie MeSH
- nifedipin farmakologie MeSH
- pohyb buněk účinky léků genetika MeSH
- proliferace buněk účinky léků genetika MeSH
- protinádorové látky imunologicky aktivní farmakologie MeSH
- pumpa pro výměnu sodíku a vápníku genetika metabolismus MeSH
- receptor erbB-2 genetika metabolismus MeSH
- regulace genové exprese u nádorů účinky léků MeSH
- RNA interference MeSH
- trastuzumab farmakologie MeSH
- triple-negativní karcinom prsu genetika metabolismus patologie MeSH
- vápníková signalizace účinky léků genetika MeSH
- Check Tag
- lidé MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
- práce podpořená grantem MeSH
- Názvy látek
- blokátory kalciových kanálů MeSH
- inositol-1,4,5-trisfosfát - receptory MeSH
- nifedipin MeSH
- protinádorové látky imunologicky aktivní MeSH
- pumpa pro výměnu sodíku a vápníku MeSH
- receptor erbB-2 MeSH
- sodium-calcium exchanger 1 MeSH Prohlížeč
- trastuzumab MeSH
