Lipocalin genes NLaz, GLaz, and Karl are evolutionarily conserved genes in Drosophila melanogaster. There are studies on lipocalin gene expression differences under diverse diet conditions, but these studies have focused mainly on age-dependent expression profiles of these genes. The main aim of our study is to determine lipocalin expression in the developmental period by nutritional manipulation with an isofemale-based design. Three larval developmental periods have been researched under normal and restricted diets. We found significant differences between lines during their developmental time-related lipocalin expression. Here, we demonstrate that upregulations in the early developmental stages of lipocalin genes under stressful conditions resulted in unaffected developmental time. The possible reason for high expression is the activation of stress signal pathways in order to buffer the harmful effects of nutritional restriction. Our data showed that the early developmental period (48-72 h) is especially crucial to tolerate the dietary stress with respect to GLaz and NLaz expression. Results of this experiment have shown that the expression profiles of lipocalin genes have line-specific pathways to nutritional stress. Their expression depends on the genetic background corresponding to development time results. Our results highlight the transcriptional changes of lipocalins associated with developmental time in larvae, developed in a dietary-restricted medium.
{"title":"Lipocalin gene expression is varied in developmental stages by larval nutritional stress in Drosophila","authors":"N. Ayhan, Pınar Güler, Banu Şebnem Önder","doi":"10.3906/biy-1604-35","DOIUrl":"https://doi.org/10.3906/biy-1604-35","url":null,"abstract":"Lipocalin genes NLaz, GLaz, and Karl are evolutionarily conserved genes in Drosophila melanogaster. There are studies on lipocalin gene expression differences under diverse diet conditions, but these studies have focused mainly on age-dependent expression profiles of these genes. The main aim of our study is to determine lipocalin expression in the developmental period by nutritional manipulation with an isofemale-based design. Three larval developmental periods have been researched under normal and restricted diets. We found significant differences between lines during their developmental time-related lipocalin expression. Here, we demonstrate that upregulations in the early developmental stages of lipocalin genes under stressful conditions resulted in unaffected developmental time. The possible reason for high expression is the activation of stress signal pathways in order to buffer the harmful effects of nutritional restriction. Our data showed that the early developmental period (48-72 h) is especially crucial to tolerate the dietary stress with respect to GLaz and NLaz expression. Results of this experiment have shown that the expression profiles of lipocalin genes have line-specific pathways to nutritional stress. Their expression depends on the genetic background corresponding to development time results. Our results highlight the transcriptional changes of lipocalins associated with developmental time in larvae, developed in a dietary-restricted medium.","PeriodicalId":23358,"journal":{"name":"Turkish Journal of Biology","volume":"41 1","pages":"178-186"},"PeriodicalIF":2.2,"publicationDate":"2017-02-20","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3906/biy-1604-35","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43300852","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
M. Revalska, V. Vassileva, G. Zehirov, S. Goormachtig, A. Iantcheva
The phytohormone auxin is a critical signal molecule, regulating fundamental processes in plant growth and development, such as shaping the root and shoot architecture, organ patterning, and nodulation. Auxin regulates plant gene expression mainly through auxin response factors (ARFs), which bind to auxin response elements in the promoter, upstream of auxin-activated genes. Here we examine and assess the function and expression pattern of a gene described as an auxin response factor, containing a DNA-binding pseudobarrel and B3 DNA-binding domains, from Medicago truncatula (MtARF-B3). For the model legume species M. truncatula, stable transgenic plants with MtARF-B3 overexpression, downregulation, and transcriptional reporters were constructed. Phenotypic and morphological evaluation of the obtained transgenic plants confirmed the important role of MtARF-B3 in general plant growth and development, modeling of root architecture, and development of seeds. Detailed histochemical and transcriptional analysis revealed expression of the gene in various stages of somatic embryogenesis, during formation of plant organs and tissues, and symbiotic nodulation. The fact that MtARF-B3 was strongly expressed in stamens and pollen grains in M. truncatula suggests that this gene could play a role in the fertility of this model legume.
{"title":"Assessment of the function and expression pattern of auxin response factor B3 in the model legume plant Medicago truncatula","authors":"M. Revalska, V. Vassileva, G. Zehirov, S. Goormachtig, A. Iantcheva","doi":"10.3906/BIY-1602-36","DOIUrl":"https://doi.org/10.3906/BIY-1602-36","url":null,"abstract":"The phytohormone auxin is a critical signal molecule, regulating fundamental processes in plant growth and development, such as shaping the root and shoot architecture, organ patterning, and nodulation. Auxin regulates plant gene expression mainly through auxin response factors (ARFs), which bind to auxin response elements in the promoter, upstream of auxin-activated genes. Here we examine and assess the function and expression pattern of a gene described as an auxin response factor, containing a DNA-binding pseudobarrel and B3 DNA-binding domains, from Medicago truncatula (MtARF-B3). For the model legume species M. truncatula, stable transgenic plants with MtARF-B3 overexpression, downregulation, and transcriptional reporters were constructed. Phenotypic and morphological evaluation of the obtained transgenic plants confirmed the important role of MtARF-B3 in general plant growth and development, modeling of root architecture, and development of seeds. Detailed histochemical and transcriptional analysis revealed expression of the gene in various stages of somatic embryogenesis, during formation of plant organs and tissues, and symbiotic nodulation. The fact that MtARF-B3 was strongly expressed in stamens and pollen grains in M. truncatula suggests that this gene could play a role in the fertility of this model legume.","PeriodicalId":23358,"journal":{"name":"Turkish Journal of Biology","volume":"41 1","pages":"66-76"},"PeriodicalIF":2.2,"publicationDate":"2017-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3906/BIY-1602-36","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"46349809","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Na Li, D. Yin, Huijie Zhang, Jinmei Xu, F. Wen, Zheng Liu, Yaozhen Chen, Ning An, Jiajia Xin, Yazhou Wang, Wen Yin, Xingbin Hu
The hematopoietic microenvironment regulates self-renewal and differentiation of hematopoietic stem cells. Mesenchymal stromal cells (MSCs) contribute to the niche and participate in supporting dendritic cell (DC) commitment in vitro and in vivo. However, due to MSCs being heterogenic, it is necessary to understand the function of the MSC subpopulation in modulating the DC commitment. The current study showed that one of the bone-related Sca1+ MSCs enhanced bone marrow cell differentiation into DCs by direct cell-to-cell contact. Furthermore, the expression of STAT3 and STAT5 genes decreased in supernatant cells after Sca1+ MSCs were cocultured with bone marrow cells. In contrast, the expression level of caspase-3 in cells increased. At the same time, the presentation of costimulatory molecules CD80/CD86 and the migration of activated DCs towards lymph nodes were augmented by Sca1+ MSCs. The results demonstrated that the Sca1+ MSC subpopulation promotes the differentiation of bone marrow cells into DCs by direct cell-to-cell contact via STAT3/STAT5 and caspase-3 signaling. Meanwhile, Sca1+ MSCs also mediate the activation and migration capabilities of DCs.
{"title":"The Sca1+ mesenchymal stromal subpopulation promotes dendritic cell commitment in the niche","authors":"Na Li, D. Yin, Huijie Zhang, Jinmei Xu, F. Wen, Zheng Liu, Yaozhen Chen, Ning An, Jiajia Xin, Yazhou Wang, Wen Yin, Xingbin Hu","doi":"10.3906/BIY-1510-81","DOIUrl":"https://doi.org/10.3906/BIY-1510-81","url":null,"abstract":"The hematopoietic microenvironment regulates self-renewal and differentiation of hematopoietic stem cells. Mesenchymal stromal cells (MSCs) contribute to the niche and participate in supporting dendritic cell (DC) commitment in vitro and in vivo. However, due to MSCs being heterogenic, it is necessary to understand the function of the MSC subpopulation in modulating the DC commitment. The current study showed that one of the bone-related Sca1+ MSCs enhanced bone marrow cell differentiation into DCs by direct cell-to-cell contact. Furthermore, the expression of STAT3 and STAT5 genes decreased in supernatant cells after Sca1+ MSCs were cocultured with bone marrow cells. In contrast, the expression level of caspase-3 in cells increased. At the same time, the presentation of costimulatory molecules CD80/CD86 and the migration of activated DCs towards lymph nodes were augmented by Sca1+ MSCs. The results demonstrated that the Sca1+ MSC subpopulation promotes the differentiation of bone marrow cells into DCs by direct cell-to-cell contact via STAT3/STAT5 and caspase-3 signaling. Meanwhile, Sca1+ MSCs also mediate the activation and migration capabilities of DCs.","PeriodicalId":23358,"journal":{"name":"Turkish Journal of Biology","volume":"41 1","pages":"58-65"},"PeriodicalIF":2.2,"publicationDate":"2017-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3906/BIY-1510-81","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42709180","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
E. Kaya, F. V. Souza, Emel Yilmaz Gökdoğan, Muammer Ceylan, M. Jenderek
* Correspondence: ergunkaya@mu.edu.tr
*通信：ergunkaya@mu.edu.tr
{"title":"Cryopreservation of citrus seed via dehydration followed by immersion in liquid nitrogen","authors":"E. Kaya, F. V. Souza, Emel Yilmaz Gökdoğan, Muammer Ceylan, M. Jenderek","doi":"10.3906/BIY-1603-92","DOIUrl":"https://doi.org/10.3906/BIY-1603-92","url":null,"abstract":"* Correspondence: ergunkaya@mu.edu.tr","PeriodicalId":23358,"journal":{"name":"Turkish Journal of Biology","volume":"41 1","pages":"242-248"},"PeriodicalIF":2.2,"publicationDate":"2017-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44618708","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
N. I. Zainudin, F. Hamzah, N. A. Kusai, Nur Syuhada Zambri, Suhaida Salleh
Fusarium ear rot is a significant disease of corn caused by several toxigenic Fusarium species including Fusarium proliferatum and Fusarium verticillioides. Forty-one Fusarium isolates were recovered from corn with Fusarium ear rot disease symptoms collected from Peninsular Malaysia. Isolates were classified into three described species known as F. proliferatum, F. verticillioides, and F. solani. Based on sexual compatibility test, four isolates from F. proliferatum (MATD-1) were crossed-fertile with tester isolate F. proliferatum D024853 (MATD-2), producing perithecia in the presence of ascospores. Meanwhile, the isolates from F. verticillioides (MATA-2) were crossed with tester isolate F. verticillioides A00149 (MATA-1), but were found producing 11 isolates with barren perithecia and three infertile isolates, whereas in 11 isolates of F. verticillioides (MATA-1), seven isolates produced barren perithecia with four nonfertile isolates. In the pathogenicity test, all isolates were found pathogenic and displayed disease symptoms with variation in severity. The highest disease severity index value was observed in F. proliferatum B68c at 4.67, which was obtained in an updated report on the mating type of F. verticillioides and F. proliferatum isolated from Fusarium ear rot disease.
{"title":"Characterization and pathogenicity of Fusarium proliferatum and Fusarium verticillioides, causal agents of Fusarium ear rot of corn","authors":"N. I. Zainudin, F. Hamzah, N. A. Kusai, Nur Syuhada Zambri, Suhaida Salleh","doi":"10.3906/BIY-1606-25","DOIUrl":"https://doi.org/10.3906/BIY-1606-25","url":null,"abstract":"Fusarium ear rot is a significant disease of corn caused by several toxigenic Fusarium species including Fusarium proliferatum and Fusarium verticillioides. Forty-one Fusarium isolates were recovered from corn with Fusarium ear rot disease symptoms collected from Peninsular Malaysia. Isolates were classified into three described species known as F. proliferatum, F. verticillioides, and F. solani. Based on sexual compatibility test, four isolates from F. proliferatum (MATD-1) were crossed-fertile with tester isolate F. proliferatum D024853 (MATD-2), producing perithecia in the presence of ascospores. Meanwhile, the isolates from F. verticillioides (MATA-2) were crossed with tester isolate F. verticillioides A00149 (MATA-1), but were found producing 11 isolates with barren perithecia and three infertile isolates, whereas in 11 isolates of F. verticillioides (MATA-1), seven isolates produced barren perithecia with four nonfertile isolates. In the pathogenicity test, all isolates were found pathogenic and displayed disease symptoms with variation in severity. The highest disease severity index value was observed in F. proliferatum B68c at 4.67, which was obtained in an updated report on the mating type of F. verticillioides and F. proliferatum isolated from Fusarium ear rot disease.","PeriodicalId":23358,"journal":{"name":"Turkish Journal of Biology","volume":"41 1","pages":"220-230"},"PeriodicalIF":2.2,"publicationDate":"2017-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3906/BIY-1606-25","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"44370287","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
T. Arasoglu, S. Derman, B. Mansuroğlu, Deniz Uzunoğlu, B. Kocyigit, B. Gümüș, T. Acar, B. Tuncer
The use of quercetin as a bioflavonoid is becoming increasingly common in food industries even though poor water solubility, instability, absorption, and permeability have limited its application. The oil-in-water single-emulsion solvent evaporation method to synthesize highly stable and soluble quercetin-encapsulated nanoparticles (NPs), in which the reaction yield, particle size, and polydispersity of the NPs are varied greatly within the process parameters of the synthesis method, has been optimized. NPs with different initial quercetin amounts were used to determine how the quercetin amount affected nanoparticle properties and antimicrobial eficiency. Listeria monocytogenes, Salmonella typhimurium, Escherichia coli, and Staphylococcus aureus were chosen as model bacteria due to their being foodborne pathogens. The results of antimicrobial activity evaluated by three different methods showed that the antimicrobial activity of both quercetin NPs and free quercetin was effective on gram-positive strains (L. monocytogenes and S. aureus). Additionally, it was detected that Q31 NPs have more effective antimicrobial activity than other synthesized quercetin nanoparticles depending on the amount of substance and release. Furthermore, on the basis of assessing the antibacterial effects by scanning electron microscopy, it was detected that bacteria cells lost their integrity and became pale with the release of cytoplasm and decomposed after treatment with Q31 NPs.
{"title":"Preparation, characterization, and enhanced antimicrobial activity: quercetin-loaded PLGA nanoparticles against foodborne pathogens","authors":"T. Arasoglu, S. Derman, B. Mansuroğlu, Deniz Uzunoğlu, B. Kocyigit, B. Gümüș, T. Acar, B. Tuncer","doi":"10.3906/BIY-1604-80","DOIUrl":"https://doi.org/10.3906/BIY-1604-80","url":null,"abstract":"The use of quercetin as a bioflavonoid is becoming increasingly common in food industries even though poor water solubility, instability, absorption, and permeability have limited its application. The oil-in-water single-emulsion solvent evaporation method to synthesize highly stable and soluble quercetin-encapsulated nanoparticles (NPs), in which the reaction yield, particle size, and polydispersity of the NPs are varied greatly within the process parameters of the synthesis method, has been optimized. NPs with different initial quercetin amounts were used to determine how the quercetin amount affected nanoparticle properties and antimicrobial eficiency. Listeria monocytogenes, Salmonella typhimurium, Escherichia coli, and Staphylococcus aureus were chosen as model bacteria due to their being foodborne pathogens. The results of antimicrobial activity evaluated by three different methods showed that the antimicrobial activity of both quercetin NPs and free quercetin was effective on gram-positive strains (L. monocytogenes and S. aureus). Additionally, it was detected that Q31 NPs have more effective antimicrobial activity than other synthesized quercetin nanoparticles depending on the amount of substance and release. Furthermore, on the basis of assessing the antibacterial effects by scanning electron microscopy, it was detected that bacteria cells lost their integrity and became pale with the release of cytoplasm and decomposed after treatment with Q31 NPs.","PeriodicalId":23358,"journal":{"name":"Turkish Journal of Biology","volume":"41 1","pages":"127-140"},"PeriodicalIF":2.2,"publicationDate":"2017-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3906/BIY-1604-80","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48823880","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
I. Kunter, E. Kandemis, Hani Alotaibi, T. Canda, E. E. Bağrıyanık
ING1 has regulatory roles in the expression of genes associated with proliferation, apoptosis, and senescence. p33(ING1b) is the most widely expressed isoform of the gene. Downregulation of its nuclear expression is involved in differentiation and pathogenesis in invasive breast carcinoma. Yet the mechanism(s) by which p33 nuclear targeting is regulated remains unknown. In this study, we analyzed human invasive breast carcinoma tissue samples by immunostaining with p33 and correlating p33 location with the presence of ERα. Our findings show the expression of p33 protein in ERα-positive tumor samples was in the nucleus alone, while the expression was mainly in the cytoplasm in ERα-negative tumor samples. Examination of the localization of p33 in the nucleus and/or cytoplasm in several different cell lines demonstrated 17β-estradiol (E2) treatment causes dramatic compartmental shift in p33 protein from the cytoplasm to the nucleus in ERα-positive MDA-66 cells. No significant differences in ERα-negative MDA-MB-231 cells in the same conditions were observed. We show for the first time nuclear localization of p33 is regulated by estradiol induction in ERα-positive breast cancer cells. These results suggest compartmental shift in p33 by ER signaling may be an important molecular event in the differentiation and pathogenesis of invasive breast cancer.
{"title":"Alteration in the subcellular location of the inhibitor of growth protein p33(ING1b) in estrogen receptor alpha positive breast carcinoma cells","authors":"I. Kunter, E. Kandemis, Hani Alotaibi, T. Canda, E. E. Bağrıyanık","doi":"10.3906/biy-1602-95","DOIUrl":"https://doi.org/10.3906/biy-1602-95","url":null,"abstract":"ING1 has regulatory roles in the expression of genes associated with proliferation, apoptosis, and senescence. p33(ING1b) is the most widely expressed isoform of the gene. Downregulation of its nuclear expression is involved in differentiation and pathogenesis in invasive breast carcinoma. Yet the mechanism(s) by which p33 nuclear targeting is regulated remains unknown. In this study, we analyzed human invasive breast carcinoma tissue samples by immunostaining with p33 and correlating p33 location with the presence of ERα. Our findings show the expression of p33 protein in ERα-positive tumor samples was in the nucleus alone, while the expression was mainly in the cytoplasm in ERα-negative tumor samples. Examination of the localization of p33 in the nucleus and/or cytoplasm in several different cell lines demonstrated 17β-estradiol (E2) treatment causes dramatic compartmental shift in p33 protein from the cytoplasm to the nucleus in ERα-positive MDA-66 cells. No significant differences in ERα-negative MDA-MB-231 cells in the same conditions were observed. We show for the first time nuclear localization of p33 is regulated by estradiol induction in ERα-positive breast cancer cells. These results suggest compartmental shift in p33 by ER signaling may be an important molecular event in the differentiation and pathogenesis of invasive breast cancer.","PeriodicalId":23358,"journal":{"name":"Turkish Journal of Biology","volume":"41 1","pages":"105-112"},"PeriodicalIF":2.2,"publicationDate":"2017-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3906/biy-1602-95","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"48051122","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A. Oral, H. Oral, Mehmet Sarimahmut, B. Cevatemre, G. Ozkaya, Şeniz Korkmaz, E. Ulukaya
The vacuolar (H+)-ATPases that pump H+ from the cytoplasm to extracellular compartments can alter the pH of the tumor microenvironment. Esomeprazole can effectively inhibit vacuolar (H+)-ATPases and may increase the effectiveness of chemotherapeutics. Therefore, we used esomeprazole in combination with cisplatin, carboplatin, paclitaxel, docetaxel, gemcitabine, and vinorelbine on the A549 nonsmall-cell lung cancer cell line. Cisplatin and carboplatin combinations with esomeprazole exhibited superior cytotoxicity compared to the other selected chemotherapeutics. Low-dose combinations of esomeprazole with either cisplatin or carboplatin resulted in synergistic interaction. We examined cytotoxic activity of these combinations with the xCELLigence real-time cytotoxicity assay and detected that esomeprazole combinations with both 100% test drug concentrations of cisplatin and carboplatin shifted the antiproliferative effects of these agents towards a cytotoxic effect in a dose-dependent manner. Cell death mode was investigated by M30 assay, Annexin-V-FITC fluorescence imaging, and determination of PARP cleavage in western blotting. The cells treated with the cisplatin and esomeprazole combination displayed characteristic features of apoptosis such as elevated M30 levels, Annexin-V staining, and PARP cleavage. In conclusion, these novel combinations resulted in higher sensitivity of tumors to chemotherapeutics, thereby warranting further in vivo experiments for proof of the concept.
{"title":"Combination of esomeprazole with chemotherapeutics results in more pronounced cytotoxic effect via apoptosis on A549 nonsmall-cell lung cancer cell line","authors":"A. Oral, H. Oral, Mehmet Sarimahmut, B. Cevatemre, G. Ozkaya, Şeniz Korkmaz, E. Ulukaya","doi":"10.3906/BIY-1606-46","DOIUrl":"https://doi.org/10.3906/BIY-1606-46","url":null,"abstract":"The vacuolar (H+)-ATPases that pump H+ from the cytoplasm to extracellular compartments can alter the pH of the tumor microenvironment. Esomeprazole can effectively inhibit vacuolar (H+)-ATPases and may increase the effectiveness of chemotherapeutics. Therefore, we used esomeprazole in combination with cisplatin, carboplatin, paclitaxel, docetaxel, gemcitabine, and vinorelbine on the A549 nonsmall-cell lung cancer cell line. Cisplatin and carboplatin combinations with esomeprazole exhibited superior cytotoxicity compared to the other selected chemotherapeutics. Low-dose combinations of esomeprazole with either cisplatin or carboplatin resulted in synergistic interaction. We examined cytotoxic activity of these combinations with the xCELLigence real-time cytotoxicity assay and detected that esomeprazole combinations with both 100% test drug concentrations of cisplatin and carboplatin shifted the antiproliferative effects of these agents towards a cytotoxic effect in a dose-dependent manner. Cell death mode was investigated by M30 assay, Annexin-V-FITC fluorescence imaging, and determination of PARP cleavage in western blotting. The cells treated with the cisplatin and esomeprazole combination displayed characteristic features of apoptosis such as elevated M30 levels, Annexin-V staining, and PARP cleavage. In conclusion, these novel combinations resulted in higher sensitivity of tumors to chemotherapeutics, thereby warranting further in vivo experiments for proof of the concept.","PeriodicalId":23358,"journal":{"name":"Turkish Journal of Biology","volume":"41 1","pages":"231-241"},"PeriodicalIF":2.2,"publicationDate":"2017-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3906/BIY-1606-46","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"42648049","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Ilgin Cagnan, F. Kaya, Fahriye Duygu Çetinkaya, A. Ozcan
Human mesenchymal stromal cells (MSCs) from different parts of the body (i.e. bone marrow, BM) have distinct cellular and molecular features including global gene expression profiles. Quantitative polymerase chain reaction is a reliable method used in the quantification of gene expression. Correct assessment of target gene expression mostly depends on the reference gene (RG) of choice. Herein, expression levels of RGs (n = 19) in adipo-/osteogenic-differentiated and control (uninduced) BM-MSCs from donors were measured using the RealTime ready Human Reference Gene Panel. Characterization of BM-MSCs was assessed using in vitro differentiation by histochemical staining and immunophenotyping of cells by flow cytometric analysis. BM-MSCs successfully differentiated into adipogenic and osteogenic lineages. When three groups were analyzed together, the NormFinder and GeNorm software programs identified PBGD as the most stable RG. GeNorm also reported G6PDH as the other stable gene. Due to their low expression, PBGD and G6PDH were not suitable RG candidates. RPLP0 had slightly lower stability but very high expression, rendering it as the best RG. When either adipogenic- or osteogenic-induced MSCs were analyzed with control BM-MSCs, the most stable RGs with high expression levels were revealed to be GAPDH and RPLP0. Even though ACTB had the highest expression, its stability was low.
{"title":"Stably expressed reference genes during differentiation of bone marrow-derived mesenchymal stromal cells","authors":"Ilgin Cagnan, F. Kaya, Fahriye Duygu Çetinkaya, A. Ozcan","doi":"10.3906/BIY-1511-93","DOIUrl":"https://doi.org/10.3906/BIY-1511-93","url":null,"abstract":"Human mesenchymal stromal cells (MSCs) from different parts of the body (i.e. bone marrow, BM) have distinct cellular and molecular features including global gene expression profiles. Quantitative polymerase chain reaction is a reliable method used in the quantification of gene expression. Correct assessment of target gene expression mostly depends on the reference gene (RG) of choice. Herein, expression levels of RGs (n = 19) in adipo-/osteogenic-differentiated and control (uninduced) BM-MSCs from donors were measured using the RealTime ready Human Reference Gene Panel. Characterization of BM-MSCs was assessed using in vitro differentiation by histochemical staining and immunophenotyping of cells by flow cytometric analysis. BM-MSCs successfully differentiated into adipogenic and osteogenic lineages. When three groups were analyzed together, the NormFinder and GeNorm software programs identified PBGD as the most stable RG. GeNorm also reported G6PDH as the other stable gene. Due to their low expression, PBGD and G6PDH were not suitable RG candidates. RPLP0 had slightly lower stability but very high expression, rendering it as the best RG. When either adipogenic- or osteogenic-induced MSCs were analyzed with control BM-MSCs, the most stable RGs with high expression levels were revealed to be GAPDH and RPLP0. Even though ACTB had the highest expression, its stability was low.","PeriodicalId":23358,"journal":{"name":"Turkish Journal of Biology","volume":"41 1","pages":"88-97"},"PeriodicalIF":2.2,"publicationDate":"2017-02-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.3906/BIY-1511-93","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"43167996","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":4,"RegionCategory":"生物学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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