A. Le, Z. Wang, X. Dai, T. Xiao, R. Zhuo, B. Zhang, Zhonglin Xiao, X. Fang
This study aimed to investigate the effect of icariin (ICA) on thin endometrium in a rat model. To this end, 6- to 8-week-old female Sprague Dawley rats (105) were randomly divided into 7 groups: untreated, vehicle-treated (lavage with NaCl), high-dose ICA (lavage with ICA at 200 mg∙kg-1∙day-1), medium-dose ICA (lavage ICA at 100 mg∙kg-1∙day-1), low-dose ICA (lavage with ICA at 50 mg∙kg-1∙day-1), sham model (injected with NaCl at uterus horn), and sample group. To induce thin endometrium, rats of all groups (except sham-model) were injected with 95% ethanol via the uterine horn. Each group underwent its respective treatment for 3 estrous cycles, after which 5 rats from each group were sacrificed, and endometrial thickness was measured. The expression of CD31, factor VIII, vascular endothelial growth factor (VEGF), cytokeratin (CK), and vimentin were detected via immunohistochemistry. The results showed that CD31, factor VIII, and VEGF were primarily expressed in the cytoplasm of endometrial and vascular epithelial cells. No difference in the expression of these factors was detected between the ICA lavage groups and the untreated groups. However, high dose ICA-treated group exhibited significantly higher expression of CD31, factor VIII, and VEGF compared to that in the low dose and vehicle-treated groups. CK and vimentin in the endometrial tissue were significantly higher in the untreated and treatment groups compared to the vehicle-treated group. This study demonstrated that ICA increases thickness of the endometrium, and it may modulate expression of VEGF, CD31, and factor VIII.
{"title":"An experimental study on the use of icariin for improving thickness of thin endometrium.","authors":"A. Le, Z. Wang, X. Dai, T. Xiao, R. Zhuo, B. Zhang, Zhonglin Xiao, X. Fang","doi":"10.4238/gmr16019126","DOIUrl":"https://doi.org/10.4238/gmr16019126","url":null,"abstract":"This study aimed to investigate the effect of icariin (ICA) on thin endometrium in a rat model. To this end, 6- to 8-week-old female Sprague Dawley rats (105) were randomly divided into 7 groups: untreated, vehicle-treated (lavage with NaCl), high-dose ICA (lavage with ICA at 200 mg∙kg-1∙day-1), medium-dose ICA (lavage ICA at 100 mg∙kg-1∙day-1), low-dose ICA (lavage with ICA at 50 mg∙kg-1∙day-1), sham model (injected with NaCl at uterus horn), and sample group. To induce thin endometrium, rats of all groups (except sham-model) were injected with 95% ethanol via the uterine horn. Each group underwent its respective treatment for 3 estrous cycles, after which 5 rats from each group were sacrificed, and endometrial thickness was measured. The expression of CD31, factor VIII, vascular endothelial growth factor (VEGF), cytokeratin (CK), and vimentin were detected via immunohistochemistry. The results showed that CD31, factor VIII, and VEGF were primarily expressed in the cytoplasm of endometrial and vascular epithelial cells. No difference in the expression of these factors was detected between the ICA lavage groups and the untreated groups. However, high dose ICA-treated group exhibited significantly higher expression of CD31, factor VIII, and VEGF compared to that in the low dose and vehicle-treated groups. CK and vimentin in the endometrial tissue were significantly higher in the untreated and treatment groups compared to the vehicle-treated group. This study demonstrated that ICA increases thickness of the endometrium, and it may modulate expression of VEGF, CD31, and factor VIII.","PeriodicalId":189314,"journal":{"name":"Genetics and molecular research : GMR","volume":"9 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2017-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"115399160","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Panax japonicus C.A. Meyer, a perennial herb belonging to the Araliaceae ginseng genus, is one of the seven rare and endangered Chinese medical herbs. By cloning the SS segment, the expression vectors pCXSN-PjSS and pCXSN-antiPjSS were constructed and introduced into Agrobactria LBA4404, which is used for engineering bacteria. Polymerase chain reaction (PCR) showed that the PjSS, antiPjSS, and Hyg were integrated in Nicotiana tabacum. Reverse transcription-PCR indicated that the PjSS was transcribed into mRNA in N. tabacum and was highly expressed, while the antiPjSS was not expressed. Detection of Ginsenoside Re content showed that transgenetic N. tabacum can increase the content of Ginsenoside Re and that anti-transgenetic N. tabacum decreased the content of Ginsenoside Re.
{"title":"Construction and transformation of expression vector containing Panax japonicus SS gene.","authors":"L. Zhang, T. T. Wang","doi":"10.4238/gmr16019341","DOIUrl":"https://doi.org/10.4238/gmr16019341","url":null,"abstract":"Panax japonicus C.A. Meyer, a perennial herb belonging to the Araliaceae ginseng genus, is one of the seven rare and endangered Chinese medical herbs. By cloning the SS segment, the expression vectors pCXSN-PjSS and pCXSN-antiPjSS were constructed and introduced into Agrobactria LBA4404, which is used for engineering bacteria. Polymerase chain reaction (PCR) showed that the PjSS, antiPjSS, and Hyg were integrated in Nicotiana tabacum. Reverse transcription-PCR indicated that the PjSS was transcribed into mRNA in N. tabacum and was highly expressed, while the antiPjSS was not expressed. Detection of Ginsenoside Re content showed that transgenetic N. tabacum can increase the content of Ginsenoside Re and that anti-transgenetic N. tabacum decreased the content of Ginsenoside Re.","PeriodicalId":189314,"journal":{"name":"Genetics and molecular research : GMR","volume":"310 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2017-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"115913291","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
P. S. D. S. Leite, T. G. Fagundes, J. A. R. Nunes, R. Parrella, N. Durães, A. T. Bruzi
Sweet sorghum has emerged as an alternative crop for ethanol yield. The breeding of this crop is performed to obtain cultivars with high ethanol yield, which necessarily requires associating favorable phenotypes for multiple traits. Therefore, the aims of this study were to investigate the association between agro-industrial traits related to ethanol yield and identify the promising genotypes considering multiple traits in sweet sorghum. For this purpose, we evaluated 45 genotypes using a 9 x 5 alpha-lattice experimental design with three replications. The traits measured were flowering time, plant height, tons of stalk per hectare, total soluble solids, tons of brix per hectare, juice extraction, total recoverable sugars, and ethanol yield. Analyses were performed after the recovery of inter-block information. The interrelation of the traits was described by genotype-by-trait biplot. For simultaneous selection, the Modified Mulamba and Mock index was used. For almost all of the agro-industrial traits, except for juice extraction, selective accuracy was above 70%. There were significant differences among genotypes for all the traits. The genotype-by-trait biplot evidenced a positive association between most of the traits related to ethanol yield, except for juice extraction, indicating the possibility of indirect selection to obtain more productive genotypes. Some genotypes proved to be promising based on the selection index, as they accumulated phenotypes favorable for the traits of interest.
{"title":"Association among agro-industrial traits and simultaneous selection in sweet sorghum.","authors":"P. S. D. S. Leite, T. G. Fagundes, J. A. R. Nunes, R. Parrella, N. Durães, A. T. Bruzi","doi":"10.4238/gmr16019318","DOIUrl":"https://doi.org/10.4238/gmr16019318","url":null,"abstract":"Sweet sorghum has emerged as an alternative crop for ethanol yield. The breeding of this crop is performed to obtain cultivars with high ethanol yield, which necessarily requires associating favorable phenotypes for multiple traits. Therefore, the aims of this study were to investigate the association between agro-industrial traits related to ethanol yield and identify the promising genotypes considering multiple traits in sweet sorghum. For this purpose, we evaluated 45 genotypes using a 9 x 5 alpha-lattice experimental design with three replications. The traits measured were flowering time, plant height, tons of stalk per hectare, total soluble solids, tons of brix per hectare, juice extraction, total recoverable sugars, and ethanol yield. Analyses were performed after the recovery of inter-block information. The interrelation of the traits was described by genotype-by-trait biplot. For simultaneous selection, the Modified Mulamba and Mock index was used. For almost all of the agro-industrial traits, except for juice extraction, selective accuracy was above 70%. There were significant differences among genotypes for all the traits. The genotype-by-trait biplot evidenced a positive association between most of the traits related to ethanol yield, except for juice extraction, indicating the possibility of indirect selection to obtain more productive genotypes. Some genotypes proved to be promising based on the selection index, as they accumulated phenotypes favorable for the traits of interest.","PeriodicalId":189314,"journal":{"name":"Genetics and molecular research : GMR","volume":"120 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2017-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"116890545","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Parkinson's disease (PD) is one of the most common neurodegenerative diseases and mainly manifests with decreasing numbers of dopaminergic neurons. Rapid eye movement (REM) sleep behavior disorder (RBD) has an incidence of 15-47% in all PD patients. Prion proteins (PrPs), which are expressed in both neurons and glial cells of the brain, are believed to be correlated with abnormal neurological functions, although their role in PD-related sleeping disorders remains unclear. We therefore investigated the expressional profiles of PrP in PD patients with RBD. Quantitative real-time polymerase chain reaction and western blotting were used to detect the mRNA and protein levels of PrP, respectively, in the cerebrospinal fluid (CSF) of PD patients with RBD, PD patients without sleeping disorder, and healthy people (N = 23 each). We investigated the correlation between the CSF PrP level and sleeping behavior in PD patients. Patients with PD complicated with RBD had significantly elevated CSF PrP expression levels (both mRNA and protein) compared with either PD patients without sleeping disorder or healthy individuals (P < 0.05 in both cases). There is elevated expression of PrP in the CSF of PD patients with RBD. This may benefit the diagnosis of PD-related RBD.
{"title":"Expression of prion protein in the cerebrospinal fluid of patients with Parkinson's disease complicated with rapid eye movement sleep behavior disorder.","authors":"W. Zhang, X. Shang, J. Peng, M. H. Zhou, W. Sun","doi":"10.4238/gmr16019022","DOIUrl":"https://doi.org/10.4238/gmr16019022","url":null,"abstract":"Parkinson's disease (PD) is one of the most common neurodegenerative diseases and mainly manifests with decreasing numbers of dopaminergic neurons. Rapid eye movement (REM) sleep behavior disorder (RBD) has an incidence of 15-47% in all PD patients. Prion proteins (PrPs), which are expressed in both neurons and glial cells of the brain, are believed to be correlated with abnormal neurological functions, although their role in PD-related sleeping disorders remains unclear. We therefore investigated the expressional profiles of PrP in PD patients with RBD. Quantitative real-time polymerase chain reaction and western blotting were used to detect the mRNA and protein levels of PrP, respectively, in the cerebrospinal fluid (CSF) of PD patients with RBD, PD patients without sleeping disorder, and healthy people (N = 23 each). We investigated the correlation between the CSF PrP level and sleeping behavior in PD patients. Patients with PD complicated with RBD had significantly elevated CSF PrP expression levels (both mRNA and protein) compared with either PD patients without sleeping disorder or healthy individuals (P < 0.05 in both cases). There is elevated expression of PrP in the CSF of PD patients with RBD. This may benefit the diagnosis of PD-related RBD.","PeriodicalId":189314,"journal":{"name":"Genetics and molecular research : GMR","volume":"105 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2017-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"124758741","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
H. Bardak, M. Gunay, Y. Ercalik, Y. Bardak, H. Ozbas, O. Bagci
Age-related macular degeneration (AMD) is the leading cause of blindness in developed countries. It is a complex disease with both genetic and environmental risk factors. To improve clinical management of this condition, it is important to develop risk assessment and prevention strategies for environmental influences, and establish a more effective treatment approach. The aim of the present study was to investigate age-related maculopathy susceptibility protein 2 (ARMS2) gene sequences among Turkish patients with exudative AMD. In addition to 39 advanced exudative AMD patients, 250 healthy individuals for whom exome sequencing data were available were included as a control group. Patients with a history of known environmental and systemic AMD risk factors were excluded. Genomic DNA was isolated from peripheral blood and analyzed using next-generation sequencing. All coding exons of the ARMS2 gene were assessed. Three different ARMS2 sequence variations (rs10490923, rs2736911, and rs10490924) were identified in both the patient and control group. Within the control group, two further ARMS2 gene variants (rs7088128 and rs36213074) were also detected. Logistic regression analysis revealed a relationship between the rs10490924 polymorphism and AMD in the Turkish population.
{"title":"Next-generation sequencing analysis of the ARMS2 gene in Turkish exudative age-related macular degeneration patients.","authors":"H. Bardak, M. Gunay, Y. Ercalik, Y. Bardak, H. Ozbas, O. Bagci","doi":"10.4238/gmr16019135","DOIUrl":"https://doi.org/10.4238/gmr16019135","url":null,"abstract":"Age-related macular degeneration (AMD) is the leading cause of blindness in developed countries. It is a complex disease with both genetic and environmental risk factors. To improve clinical management of this condition, it is important to develop risk assessment and prevention strategies for environmental influences, and establish a more effective treatment approach. The aim of the present study was to investigate age-related maculopathy susceptibility protein 2 (ARMS2) gene sequences among Turkish patients with exudative AMD. In addition to 39 advanced exudative AMD patients, 250 healthy individuals for whom exome sequencing data were available were included as a control group. Patients with a history of known environmental and systemic AMD risk factors were excluded. Genomic DNA was isolated from peripheral blood and analyzed using next-generation sequencing. All coding exons of the ARMS2 gene were assessed. Three different ARMS2 sequence variations (rs10490923, rs2736911, and rs10490924) were identified in both the patient and control group. Within the control group, two further ARMS2 gene variants (rs7088128 and rs36213074) were also detected. Logistic regression analysis revealed a relationship between the rs10490924 polymorphism and AMD in the Turkish population.","PeriodicalId":189314,"journal":{"name":"Genetics and molecular research : GMR","volume":"11 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2017-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"127275227","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Guangliang Gao, C. Wang, X. Zhao, H. Wang, Q. Li, J. Li, K. Zhang, H. Zhong, Q. Wang
Chicken meat quality is becoming increasingly important among breeders and consumers. To understand the effect of feeding conditions on chicken meat quality, we investigated the profiles of genes expressed in chicken breast muscle. Using RNA sequencing, we identified 336, 321, and 387 differentially expressed genes among Chengkou, Daninghe, and Qingjiaoma chickens under scatter- and captivity-feeding conditions. Twenty-two genes differentially expressed between different feeding conditions were shown to be common among the three breeds. Seven of these genes were assessed by real-time quantitative PCR, which confirmed the findings of RNA sequencing and suggested that the results were viable. The differentially expressed genes showed enrichment for a series of significant pathways, including energy metabolism, xenobiotics biodegradation and metabolism, and the immune system. These results provide a solid foundation for elucidating the molecular mechanisms underlying chicken meat quality.
{"title":"Effects of feeding conditions on gene expression in chicken breast muscle.","authors":"Guangliang Gao, C. Wang, X. Zhao, H. Wang, Q. Li, J. Li, K. Zhang, H. Zhong, Q. Wang","doi":"10.4238/gmr16019119","DOIUrl":"https://doi.org/10.4238/gmr16019119","url":null,"abstract":"Chicken meat quality is becoming increasingly important among breeders and consumers. To understand the effect of feeding conditions on chicken meat quality, we investigated the profiles of genes expressed in chicken breast muscle. Using RNA sequencing, we identified 336, 321, and 387 differentially expressed genes among Chengkou, Daninghe, and Qingjiaoma chickens under scatter- and captivity-feeding conditions. Twenty-two genes differentially expressed between different feeding conditions were shown to be common among the three breeds. Seven of these genes were assessed by real-time quantitative PCR, which confirmed the findings of RNA sequencing and suggested that the results were viable. The differentially expressed genes showed enrichment for a series of significant pathways, including energy metabolism, xenobiotics biodegradation and metabolism, and the immune system. These results provide a solid foundation for elucidating the molecular mechanisms underlying chicken meat quality.","PeriodicalId":189314,"journal":{"name":"Genetics and molecular research : GMR","volume":"86 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2017-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"129148989","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
N. Hasan, M. Rafii, H. Abdul Rahim, S. A. Nusaibah, N. Mazlan, S. Abdullah
Rice (Oryza sativa L.) blast disease is one of the most destructive rice diseases in the world. The fungal pathogen, Magnaporthe oryzae, is the causal agent of rice blast disease. Development of resistant cultivars is the most preferred method to achieve sustainable rice production. However, the effectiveness of resistant cultivars is hindered by the genetic plasticity of the pathogen genome. Therefore, information on genetic resistance and virulence stability are vital to increase our understanding of the molecular basis of blast disease resistance. The present study set out to elucidate the resistance pattern and identify potential simple sequence repeat markers linked with rice blast disease. A backcross population (BC2F1), derived from crossing MR264 and Pongsu Seribu 2 (PS2), was developed using marker-assisted backcross breeding. Twelve microsatellite markers carrying the blast resistance gene clearly demonstrated a polymorphic pattern between both parental lines. Among these, two markers, RM206 and RM5961, located on chromosome 11 exhibited the expected 1:1 testcross ratio in the BC2F1 population. The 195 BC2F1 plants inoculated against M. oryzae pathotype P7.2 showed a significantly different distribution in the backcrossed generation and followed Mendelian segregation based on a single-gene model. This indicates that blast resistance in PS2 is governed by a single dominant gene, which is linked to RM206 and RM5961 on chromosome 11. The findings presented in this study could be useful for future blast resistance studies in rice breeding programs.
{"title":"Genetic analysis and identification of SSR markers associated with rice blast disease in a BC2F1 backcross population.","authors":"N. Hasan, M. Rafii, H. Abdul Rahim, S. A. Nusaibah, N. Mazlan, S. Abdullah","doi":"10.4238/gmr16019280","DOIUrl":"https://doi.org/10.4238/gmr16019280","url":null,"abstract":"Rice (Oryza sativa L.) blast disease is one of the most destructive rice diseases in the world. The fungal pathogen, Magnaporthe oryzae, is the causal agent of rice blast disease. Development of resistant cultivars is the most preferred method to achieve sustainable rice production. However, the effectiveness of resistant cultivars is hindered by the genetic plasticity of the pathogen genome. Therefore, information on genetic resistance and virulence stability are vital to increase our understanding of the molecular basis of blast disease resistance. The present study set out to elucidate the resistance pattern and identify potential simple sequence repeat markers linked with rice blast disease. A backcross population (BC2F1), derived from crossing MR264 and Pongsu Seribu 2 (PS2), was developed using marker-assisted backcross breeding. Twelve microsatellite markers carrying the blast resistance gene clearly demonstrated a polymorphic pattern between both parental lines. Among these, two markers, RM206 and RM5961, located on chromosome 11 exhibited the expected 1:1 testcross ratio in the BC2F1 population. The 195 BC2F1 plants inoculated against M. oryzae pathotype P7.2 showed a significantly different distribution in the backcrossed generation and followed Mendelian segregation based on a single-gene model. This indicates that blast resistance in PS2 is governed by a single dominant gene, which is linked to RM206 and RM5961 on chromosome 11. The findings presented in this study could be useful for future blast resistance studies in rice breeding programs.","PeriodicalId":189314,"journal":{"name":"Genetics and molecular research : GMR","volume":"24 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2017-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"126946273","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Cervical carcinoma is a life-threatening illness posing considerable danger to women's health. microRNAs (miRNAs) have been shown to regulate multiple cellular events, including growth and proliferation, and miR-187 is thought to regulate the growth and apoptosis of certain cell types. Our study focused on the influence of miR-187 on the growth, proliferation, and apoptosis of SiHa cervical carcinoma cells, and explored the mechanism behind its pro-apoptotic effect. miR-187 and control (scrambled) miRNA were synthesized with a standard protocol and lipofected into SiHa cells. Thiazolyl blue tetrazolium bromide assays and tests of caspase-3 activity were then performed to examine growth, proliferation, and apoptosis by flow cytometry. Small interfering RNA (siRNA) and an expression plasmid were synthesized for inhibition and overexpression of Bcl-2, respectively, and following their transfection, western blotting was used to examine Bcl-2 protein levels. Compared to transfection with control miRNA, miR-187 significantly reduced SiHa cell growth and decreased Bcl-2 expression. Increased translocation of phosphatidylserine and activation of caspase-3 were observed in miR-187-transfected cells. Moreover, inhibition of Bcl-2 enhanced the pro-apoptotic effect of this miRNA, while Bcl-2 overexpression had the opposite effect. miR-187 inhibits the growth and proliferation of SiHa cells, and induces their apoptosis via downregulation of Bcl-2. Bcl-2 represents a potential therapeutic target for cervical carcinoma.
{"title":"miR-187 induces apoptosis of SiHa cervical carcinoma cells by downregulating Bcl-2.","authors":"C. He, Jun Yang","doi":"10.4238/gmr16018969","DOIUrl":"https://doi.org/10.4238/gmr16018969","url":null,"abstract":"Cervical carcinoma is a life-threatening illness posing considerable danger to women's health. microRNAs (miRNAs) have been shown to regulate multiple cellular events, including growth and proliferation, and miR-187 is thought to regulate the growth and apoptosis of certain cell types. Our study focused on the influence of miR-187 on the growth, proliferation, and apoptosis of SiHa cervical carcinoma cells, and explored the mechanism behind its pro-apoptotic effect. miR-187 and control (scrambled) miRNA were synthesized with a standard protocol and lipofected into SiHa cells. Thiazolyl blue tetrazolium bromide assays and tests of caspase-3 activity were then performed to examine growth, proliferation, and apoptosis by flow cytometry. Small interfering RNA (siRNA) and an expression plasmid were synthesized for inhibition and overexpression of Bcl-2, respectively, and following their transfection, western blotting was used to examine Bcl-2 protein levels. Compared to transfection with control miRNA, miR-187 significantly reduced SiHa cell growth and decreased Bcl-2 expression. Increased translocation of phosphatidylserine and activation of caspase-3 were observed in miR-187-transfected cells. Moreover, inhibition of Bcl-2 enhanced the pro-apoptotic effect of this miRNA, while Bcl-2 overexpression had the opposite effect. miR-187 inhibits the growth and proliferation of SiHa cells, and induces their apoptosis via downregulation of Bcl-2. Bcl-2 represents a potential therapeutic target for cervical carcinoma.","PeriodicalId":189314,"journal":{"name":"Genetics and molecular research : GMR","volume":"50 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2017-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"114096309","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Bone desensitization after mechanical loading is essential for bone to adapt to its mechanical environment. However, the desensitization mechanism is unknown. Previous studies suggest that G protein-coupled receptors (GPCRs), including P2Y and parathyroid hormone receptors, play important roles in osteoblast mechanobiology. Thus, for the present research, we examined the role of G protein-coupled receptor kinase 2 (GRK2) in osteoblast desensitization after exposure to mechanical stimulation. We first showed the existence of osteoblast desensitization after mechanical stimulation based on cytosol Ca2+ and phosphorylated ERK1/2 activities, detected using a fluorescent Ca2+-sensitive dye and western blotting, respectively. We then demonstrated that GRK2 overexpression in MC3T3-E1 cells inhibits flow-induced ERK1/2 phosphorylation, while siRNA knockdown of GRK2 enhances ERK1/2 phosphorylation. Additionally, we found that GRK2 overexpression in MC3T3-E1 cells inhibits cyclooxygenase-2 mRNA expression in the short term and alkaline phosphatase activity in the long term. More importantly, we discovered that GRK2 translocated to the cell membrane shortly after flow stimulation - a step necessary for GPCR desensitization. Previously, we have demonstrated that P2Y2 purinergic receptors, one type of GPCRs, are involved in various flow-induced osteoblastic responses. In this research, we also showed that GRK2 overexpression does not affect ATP release. Accordingly, GRK2 is able to inhibit flow-induced osteoblast responses possibly through desensitizing P2Y2 receptors.
{"title":"GRK2 desensitizes flow-induced responses in osteoblasts.","authors":"Yanghui Xing, Y. Gu, X. Shan, L. Wang, J. You","doi":"10.4238/gmr16019363","DOIUrl":"https://doi.org/10.4238/gmr16019363","url":null,"abstract":"Bone desensitization after mechanical loading is essential for bone to adapt to its mechanical environment. However, the desensitization mechanism is unknown. Previous studies suggest that G protein-coupled receptors (GPCRs), including P2Y and parathyroid hormone receptors, play important roles in osteoblast mechanobiology. Thus, for the present research, we examined the role of G protein-coupled receptor kinase 2 (GRK2) in osteoblast desensitization after exposure to mechanical stimulation. We first showed the existence of osteoblast desensitization after mechanical stimulation based on cytosol Ca2+ and phosphorylated ERK1/2 activities, detected using a fluorescent Ca2+-sensitive dye and western blotting, respectively. We then demonstrated that GRK2 overexpression in MC3T3-E1 cells inhibits flow-induced ERK1/2 phosphorylation, while siRNA knockdown of GRK2 enhances ERK1/2 phosphorylation. Additionally, we found that GRK2 overexpression in MC3T3-E1 cells inhibits cyclooxygenase-2 mRNA expression in the short term and alkaline phosphatase activity in the long term. More importantly, we discovered that GRK2 translocated to the cell membrane shortly after flow stimulation - a step necessary for GPCR desensitization. Previously, we have demonstrated that P2Y2 purinergic receptors, one type of GPCRs, are involved in various flow-induced osteoblastic responses. In this research, we also showed that GRK2 overexpression does not affect ATP release. Accordingly, GRK2 is able to inhibit flow-induced osteoblast responses possibly through desensitizing P2Y2 receptors.","PeriodicalId":189314,"journal":{"name":"Genetics and molecular research : GMR","volume":"36 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2017-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"115319608","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
B. M. Silva, A. Rossi, A. V. Tiago, K. Schmitt, J. F. E. Dardengo, Sérgio Alessandro Machado Souza
Cajazeira (Spondias mombin L.), of the family Anacardiaceae, is a species of fruit tree found in the Amazon region with fruits that have excellent prospects for commercial use. We aimed to evaluate the genetic diversity within and among natural populations of S. mombin, with natural occurrence in northern Mato Grosso State, by using inter-simple sequence repeat (ISSR) markers. Overall, 126 individuals were evaluated from three populations located Alta Floresta (AFL) 42, Marcelândia (MAR) 41, and Nova Bandeirantes (NBA) 43. The individuals were genotyped with 14 ISSR primers, which amplified 99 fragments. All markers, with the exception of DiGA3'A, presented a polymorphic information content above 0.25, and thus, are recommended for diversity analyses in S. mombin. Genetic diversity of the AFL [Nei's diversity (H) = 0.2430 and Shannon index (I) = 0.3547] and MAR (H = 0.2062 and I = 0.2993) populations was higher when compared to the NBA population, which presented the lowest genetic diversity (H = 0.2002 and I = 0.2957). Analysis of molecular variance showed that 77.38% of the total genetic variation is found within populations while 22.62% is found among populations. AFL and NBA are genetically the most similar populations and also the closest "Structure" revealed genetic diversity among the genotypes of each population. As there is genetic variability in both populations, and there are no genetically identical individuals, both populations can be a source of genotypes for germplasm banks and for future commercial fruitful plantations S. mombin.
{"title":"Genetic diversity of Cajazeira (Spondias mombin L.) in three geographic regions.","authors":"B. M. Silva, A. Rossi, A. V. Tiago, K. Schmitt, J. F. E. Dardengo, Sérgio Alessandro Machado Souza","doi":"10.4238/gmr16018946","DOIUrl":"https://doi.org/10.4238/gmr16018946","url":null,"abstract":"Cajazeira (Spondias mombin L.), of the family Anacardiaceae, is a species of fruit tree found in the Amazon region with fruits that have excellent prospects for commercial use. We aimed to evaluate the genetic diversity within and among natural populations of S. mombin, with natural occurrence in northern Mato Grosso State, by using inter-simple sequence repeat (ISSR) markers. Overall, 126 individuals were evaluated from three populations located Alta Floresta (AFL) 42, Marcelândia (MAR) 41, and Nova Bandeirantes (NBA) 43. The individuals were genotyped with 14 ISSR primers, which amplified 99 fragments. All markers, with the exception of DiGA3'A, presented a polymorphic information content above 0.25, and thus, are recommended for diversity analyses in S. mombin. Genetic diversity of the AFL [Nei's diversity (H) = 0.2430 and Shannon index (I) = 0.3547] and MAR (H = 0.2062 and I = 0.2993) populations was higher when compared to the NBA population, which presented the lowest genetic diversity (H = 0.2002 and I = 0.2957). Analysis of molecular variance showed that 77.38% of the total genetic variation is found within populations while 22.62% is found among populations. AFL and NBA are genetically the most similar populations and also the closest \"Structure\" revealed genetic diversity among the genotypes of each population. As there is genetic variability in both populations, and there are no genetically identical individuals, both populations can be a source of genotypes for germplasm banks and for future commercial fruitful plantations S. mombin.","PeriodicalId":189314,"journal":{"name":"Genetics and molecular research : GMR","volume":"2 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2017-01-23","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"131801529","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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