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Analysis of the cytochrome c oxidase subunit II (COX2) gene in giant panda, Ailuropoda melanoleuca. 大熊猫细胞色素c氧化酶亚基II (COX2)基因分析。
Pub Date : 2017-01-23 DOI: 10.4238/gmr16019158
S. Ling, Y. Zhu, D. Lan, D. S. Li, H. Pang, Y. Wang, D. Li, R. Wei, H. Zhang, C. D. Wang, Y. D. Hu
The giant panda, Ailuropoda melanoleuca (Ursidae), has a unique bamboo-based diet; however, this low-energy intake has been sufficient to maintain the metabolic processes of this species since the fourth ice age. As mitochondria are the main sites for energy metabolism in animals, the protein-coding genes involved in mitochondrial respiratory chains, particularly cytochrome c oxidase subunit II (COX2), which is the rate-limiting enzyme in electron transfer, could play an important role in giant panda metabolism. Therefore, the present study aimed to isolate, sequence, and analyze the COX2 DNA from individuals kept at the Giant Panda Protection and Research Center, China, and compare these sequences with those of the other Ursidae family members. Multiple sequence alignment showed that the COX2 gene had three point mutations that defined three haplotypes, with 60% of the sequences corresponding to haplotype I. The neutrality tests revealed that the COX2 gene was conserved throughout evolution, and the maximum likelihood phylogenetic analysis, using homologous sequences from other Ursidae species, showed clustering of the COX2 sequences of giant pandas, suggesting that this gene evolved differently in them.
大熊猫(Ailuropoda melanoleuca,熊科)以竹子为主食;然而,自第四次冰河期以来,这种低能量摄入足以维持该物种的代谢过程。由于线粒体是动物能量代谢的主要部位,参与线粒体呼吸链的蛋白质编码基因,特别是电子转移的限速酶细胞色素c氧化酶亚基II (COX2)可能在大熊猫的代谢中发挥重要作用。因此,本研究旨在从中国大熊猫保护研究中心保存的个体中分离、测序和分析COX2 DNA,并将这些序列与其他熊科成员的序列进行比较。多重序列比对显示COX2基因有3个点突变,定义了3个单倍型,其中60%的序列对应于单倍型i。中立性检验表明COX2基因在整个进化过程中是保守的,使用其他熊科物种的同源序列进行最大似然系统发育分析,发现大熊猫的COX2序列具有聚类性,表明该基因在它们中进化不同。
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引用次数: 3
Analysis of specific serum markers of colon carcinoma using a Bhattacharyya-based support vector machine. 基于bhattacharya的支持向量机分析结肠癌特异性血清标志物。
Pub Date : 2017-01-23 DOI: 10.4238/gmr16019521
Wenyi Yang, G. Shi, Liping Wu, Shutang Wei, Y. Huang, Lixia Tan, R. Yang, Chunxiao Yan, E. Guo, Hangyu Wang, J. Tong, Y. Dong, Dazheng Han
We aimed to evaluate the specificity of 12 tumor markers related to colon carcinoma and identify the most sensitive index. Bhattacharyya distance was used to evaluate the index. Then, different index combinations were used to establish a support vector machine (SVM) diagnosis model of malignant colon carcinoma. The accuracy of the model was checked. High accuracy was assumed to indicate the high specificity of the index. The Bhattacharyya distances of carcinoembryonic antigen, neuron-specific enolase, alpha-feto protein, and CA724 were the largest, and those of CYFRA21-І, CA125, and UGT1A83 were the second largest. The specificity of the combination of the above seven indexes was higher than that of other combinations, and the accuracy of the established SVM identification model was high. Using Bhattacharyya distance detection and establishing an SVM model based on different serum marker combinations can increase diagnostic accuracy, providing a theoretical basis for application of mathematical models in cancer diagnosis.
我们的目的是评估12种与结肠癌相关的肿瘤标志物的特异性,找出最敏感的指标。采用巴塔查里亚距离评价该指标。然后,采用不同的指标组合,建立了支持向量机(SVM)恶性结肠癌诊断模型。对模型的准确性进行了检验。假设高准确度表明该指数的高特异性。癌胚抗原、神经元特异性烯醇化酶、甲胎蛋白和CA724的Bhattacharyya距离最大,CYFRA21-І、CA125和UGT1A83的Bhattacharyya距离次之。以上7个指标组合的特异性高于其他组合,所建立的SVM识别模型准确率较高。采用Bhattacharyya距离检测,建立基于不同血清标志物组合的SVM模型,可提高诊断准确率,为数学模型在癌症诊断中的应用提供理论依据。
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引用次数: 2
Cytotoxicity evaluation of the whole protein extract from Bar-transgenic rice on Mus musculus lymphocytes. bar转基因水稻全蛋白提取物对小家鼠淋巴细胞的细胞毒性评价。
Pub Date : 2017-01-23 DOI: 10.4238/gmr16019331
X. B. Lu
With the expanding demand for genetically modified (GM) rice, its safety evaluation is of great significance. Therefore, this study was carried out to assess the acute cytotoxicity of the whole protein extract from GM rice Bar68-1 in Mus musculus lymphocytes in vitro. Cell viability was determined by Cell Counting Kit-8 (CCK-8) and Neutral Red Uptake (NRU) tests. CCK-8 tests was carried out according to the manufacturer's instructions. Cell dehydrogenase (catalytic redox enzymes) activity was spectrophotometrically determined at 450 nm. The tests result were recorded immediately. NRU tests were completed under yellow light in a dark room according to an improved protocol. Lysosomal uptake of neutral red was spectrophotometrically determined at 540 nm and the results were recorded immediately. The results showed that the survival rate of M. musculus lymphocytes in the positive control group was significantly less than in the blank control group (P < 0.05). Moreover, an exposure- time-effect relationship was observed in the positive control group with CCK-8 and NRU tests. There was no significant difference in survival rate between GM rice Bar68-1group and non-GM rice D68 group (P > 0.05). The GM rice Bar68-1 group also did not show a higher survival rate than non-GM rice D68 group (P > 0.05). These results suggested that the whole protein extract from Bar68-1 and D68 were equivalent in their cytotoxicity, and GM rice Bar68-1 had no acute cytotoxic effect on M. musculus lymphocytes in vitro.
随着人们对转基因水稻需求的不断扩大,转基因水稻的安全性评价具有重要意义。因此,本研究对转基因水稻全蛋白提取物Bar68-1体外对小家鼠淋巴细胞的急性细胞毒性进行了研究。通过细胞计数试剂盒-8 (CCK-8)和中性红色摄取(NRU)试验测定细胞活力。CCK-8测试是根据制造商的说明进行的。用分光光度法测定细胞脱氢酶(催化氧化还原酶)活性。测试结果记录。NRU测试在暗室黄灯下按照改进的方案完成。在540 nm处分光光度法测定溶酶体对中性红的摄取,并立即记录结果。结果显示,阳性对照组肌支原体淋巴细胞存活率显著低于空白对照组(P < 0.05)。在CCK-8和NRU测试中,阳性对照组存在暴露-时间效应关系。转基因水稻bar68 -1组与非转基因水稻D68组的成活率差异不显著(P > 0.05)。转基因水稻Bar68-1集团也没有表现出更高的存活率比非转基因大米D68组(P > 0.05)。这些结果表明,Bar68-1全蛋白提取物和D68的细胞毒性相当,转基因水稻Bar68-1对体外肌支原体淋巴细胞无急性细胞毒作用。
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引用次数: 1
Correlation-based linear discriminant classification for gene expression data. 基于相关性的基因表达数据线性判别分类。
Pub Date : 2017-01-23 DOI: 10.4238/gmr16019357
M. Pan, J. Zhang
Microarray gene expression technology provides a systematic approach to patient classification. However, microarray data pose a great computational challenge owing to their large dimensionality, small sample sizes, and potential correlations among genes. A recent study has shown that gene-gene correlations have a positive effect on the accuracy of classification models, in contrast to some previous results. In this study, a recently developed correlation-based classifier, the ensemble of random subspace (RS) Fisher linear discriminants (FLDs), was utilized. The impact of gene-gene correlations on the performance of this classifier and other classifiers was studied using simulated datasets and real datasets. A cross-validation framework was used to evaluate the performance of each classifier using the simulated datasets or real datasets, and misclassification rates (MRs) were computed. Using the simulated data, the average MRs of the correlation-based classifiers decreased as the correlations increased when there were more correlated genes. Using real data, the correlation-based classifiers outperformed the non-correlation-based classifiers, especially when the gene-gene correlations were high. The ensemble RS-FLD classifier is a potential state-of-the-art computational method. The correlation-based ensemble RS-FLD classifier was effective and benefited from gene-gene correlations, particularly when the correlations were high.
微阵列基因表达技术为患者分类提供了一种系统的方法。然而,微阵列数据由于其大维度、小样本量和基因之间的潜在相关性而带来了巨大的计算挑战。最近的一项研究表明,与之前的一些结果相反,基因-基因相关性对分类模型的准确性有积极的影响。在本研究中,使用了一种最新发展的基于相关的分类器——随机子空间(RS) Fisher线性判别器(FLDs)。利用模拟数据集和真实数据集研究了基因-基因相关性对该分类器和其他分类器性能的影响。使用交叉验证框架评估每个分类器使用模拟数据集或真实数据集的性能,并计算误分类率(MRs)。使用模拟数据,当有更多的相关基因时,基于相关性的分类器的平均MRs随着相关性的增加而降低。使用真实数据,基于相关性的分类器优于非基于相关性的分类器,特别是当基因-基因相关性较高时。集成RS-FLD分类器是一种潜在的先进计算方法。基于相关性的集合RS-FLD分类器是有效的,并受益于基因-基因相关性,特别是当相关性高时。
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引用次数: 3
An assessment of the correlation between early postinfarction pyramidal tract Wallerian degeneration and nerve function recovery using diffusion tensor imaging. 弥散张量成像评价梗死后早期锥体束沃勒氏变性与神经功能恢复的相关性。
Pub Date : 2017-01-23 DOI: 10.4238/gmr16019035
A. Guo, F. Hao, L. Liu, B. Wang, X. Jiang
This study aimed to evaluate the clinical significance of diffusion tensor imaging (DTI) in the early diagnosis of pyramidal tract Wallerian degeneration (WD) and assessment of neurological recovery following cerebral infarction. This study included 23 patients with acute cerebral infarction and 10 healthy adult controls. All participants underwent both magnetic resonance imaging (MRI) and DTI scans. DTI images were analyzed using the Functional MRI of the Brain Software Library to determine the regions of interest (ROI) and obtain the mean diffusivity (MD) and fractional anisotropy (FA) value for each ROI. The correlation between FA or MD and postinfarction functional recovery of the nervous system was further analyzed to assess the feasibility of using a DTI scan in the evaluation of functional recovery of the nervous system in patients with cerebral infarction. DTI may be useful in detecting signals of early postinfarction pyramidal tract WD and is useful for the evaluation of postinfarction neurological recovery. Cerebral lesions were detected using MRI in all patients. It was found that in some patients, the FA value of the ipsilateral pyramidal tract on DTI was decreased as early as day 3 after the onset of infarction and in all patients by day 7. Subsequent correlation studies showed that the FA value of the ipsilateral pyramidal tract on day 13 was negatively correlated with the National Institutes of Health Stroke Scale score, but positively correlated with the Barthel Index, motricity index, and modified Rankin Scale scores.
本研究旨在探讨弥散张量成像(DTI)在脑梗死后锥体束沃勒氏变性(WD)早期诊断及神经功能恢复评估中的临床意义。本研究包括23例急性脑梗死患者和10例健康成人对照。所有参与者都接受了磁共振成像(MRI)和DTI扫描。使用脑软件库的功能MRI对DTI图像进行分析,确定感兴趣区域(ROI),并获得每个ROI的平均扩散率(MD)和分数各向异性(FA)值。进一步分析FA或MD与脑梗死后神经系统功能恢复的相关性,以评估使用DTI扫描评估脑梗死患者神经系统功能恢复的可行性。DTI可用于检测梗死后早期锥体束WD信号,并可用于评价梗死后神经系统恢复情况。所有患者均行MRI检查脑病变。结果发现,部分患者的同侧锥体束FA值早在梗死发生后第3天就下降,所有患者的FA值均在第7天下降。随后的相关研究显示,第13天同侧锥体束FA值与美国国立卫生研究院卒中量表评分呈负相关,而与Barthel指数、运动力指数和改良Rankin量表评分呈正相关。
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引用次数: 6
Genetic diversity of Casearia sylvestris populations in remnants of the Atlantic Forest. 大西洋森林遗迹中西尔维斯特Casearia sylvestris种群的遗传多样性。
Pub Date : 2017-01-23 DOI: 10.4238/gmr16019105
F. L. Araujo, Marcos Vinicius B M Siqueira, C. Grando, João Paulo Gomes Viana, J. B. Pinheiro, A. Alves-Pereira, J. B. Campos, P. Brancalion, M. Zucchi
Guaçatonga (Casearia sylvestris) is a native plant of the Atlantic Forest, with high medicinal potential and relevance for reforestation programs. The aim of this study was to characterize, with microsatellite markers, two populations of C. sylvestris from remaining areas of the Atlantic Forest in the State of São Paulo. High allelic variation was found in both populations (NA = 101 and 117; AR = 12.5 and 14.4), although with high endogamy coefficients (f = 0.640 and 0.363). Estimates of genetic structure suggested the presence of considerable genetic divergence between the populations (FST = 0.103); however, there was no spatial genetic structure within the populations. Genetic divergence may have occurred due to decreased gene flow between the fragmented populations as the result of deforestation. The results of this study demonstrate the importance of genetic diversity and its characterization in native plants within remaining forest areas for the management and restoration of such areas.
瓜帕拉通加(Casearia sylvestris)是大西洋森林的一种本土植物,具有很高的药用潜力和重新造林计划的相关性。本研究的目的是利用微卫星标记对圣保罗州大西洋森林剩余地区的两个C. sylvestris种群进行特征分析。两个群体等位基因变异均较高(NA = 101和117);AR = 12.5和14.4),但内婚制系数较高(f = 0.640和0.363)。遗传结构估计表明种群之间存在相当大的遗传差异(FST = 0.103);但群体间不存在空间遗传结构。遗传分化的发生可能是由于森林砍伐导致碎片化种群之间的基因流动减少。本研究的结果表明,残存林区原生植物的遗传多样性及其特征对这些地区的管理和恢复具有重要意义。
{"title":"Genetic diversity of Casearia sylvestris populations in remnants of the Atlantic Forest.","authors":"F. L. Araujo, Marcos Vinicius B M Siqueira, C. Grando, João Paulo Gomes Viana, J. B. Pinheiro, A. Alves-Pereira, J. B. Campos, P. Brancalion, M. Zucchi","doi":"10.4238/gmr16019105","DOIUrl":"https://doi.org/10.4238/gmr16019105","url":null,"abstract":"Guaçatonga (Casearia sylvestris) is a native plant of the Atlantic Forest, with high medicinal potential and relevance for reforestation programs. The aim of this study was to characterize, with microsatellite markers, two populations of C. sylvestris from remaining areas of the Atlantic Forest in the State of São Paulo. High allelic variation was found in both populations (NA = 101 and 117; AR = 12.5 and 14.4), although with high endogamy coefficients (f = 0.640 and 0.363). Estimates of genetic structure suggested the presence of considerable genetic divergence between the populations (FST = 0.103); however, there was no spatial genetic structure within the populations. Genetic divergence may have occurred due to decreased gene flow between the fragmented populations as the result of deforestation. The results of this study demonstrate the importance of genetic diversity and its characterization in native plants within remaining forest areas for the management and restoration of such areas.","PeriodicalId":189314,"journal":{"name":"Genetics and molecular research : GMR","volume":"167 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":"122624462","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}
引用次数: 5
Protective role of +294 T/C (rs2016520) polymorphism of PPARD in Mexican patients with colorectal cancer. PPARD +294 T/C (rs2016520)多态性在墨西哥结直肠癌患者中的保护作用
Pub Date : 2017-01-23 DOI: 10.4238/gmr16019324
M. A. Rosales-Reynoso, L. Wence-Chávez, A. R. Arredondo-Valdez, S. Dumois-Petersen, Patricio Barros-Núñez, M. Gallegos-Arreola, S. Flores-Martínez, José Sánchez-Corona
PPARD encodes for peroxisome proliferator-activated receptor delta, which plays a significant role in controlling lipid metabolism, atherosclerosis, inflammation, cancer growth, progression, and apoptosis. Accumulated evidence suggests that the polymorphism rs2016520 in PPARD is associated with lipid metabolism, obesity, metabolic syndrome, and type 2 diabetes mellitus. The aim of this study was to determine whether the single nucleotide polymorphism +294T/C (rs2016520) in PPARD is associated with colorectal cancer (CRC) in the Mexican population. Genomic DNA from 178 CRC patients and 97 healthy blood donors was analyzed. The polymorphism was identified by the polymerase chain reaction-restriction fragment length polymorphism method. Results demonstrated that patients with the T/C genotype for the +294T/C (rs2016520) polymorphism present a protective role against CRC [odds ratio (OR) = 0.39; 95% confidence interval (CI) = 0.22-0.69; P = 0.0008]. This association was also evident for the T/C genotype in the stratified analysis by tumor-node-metastasis stages I+II (OR = 0.26, P = 0.0332) and III+IV (OR = 0.44, P = 0.0067). However, in the stratified analysis by tumor location, we observed an increased risk of rectal cancer (OR = 7.57, P = 0.0403) vs colon cancer (OR = 4.87, P = 0.234) in patients carrying the C/C genotype and under the dominant and recessive models of inheritance. In conclusion, for the first time, the association between the +294T/C (rs2016520) polymorphism and colorectal cancer has been studied in Mexican patients. Our results reveal that variations in PPARD may play a significant role in genetic susceptibility to colorectal cancer.
PPARD编码过氧化物酶体增殖物激活受体,在控制脂质代谢、动脉粥样硬化、炎症、癌症生长、进展和细胞凋亡中起重要作用。越来越多的证据表明PPARD中rs2016520多态性与脂质代谢、肥胖、代谢综合征和2型糖尿病有关。本研究的目的是确定PPARD中单核苷酸多态性+294T/C (rs2016520)是否与墨西哥人群的结直肠癌(CRC)相关。分析了178例结直肠癌患者和97名健康献血者的基因组DNA。采用聚合酶链反应-限制性片段长度多态性法鉴定其多态性。结果表明,T/C基因型+294T/C (rs2016520)多态性的患者对CRC具有保护作用[优势比(OR) = 0.39;95%置信区间(CI) = 0.22-0.69;P = 0.0008]。在肿瘤淋巴结转移分期I+II (OR = 0.26, P = 0.0332)和III+IV (OR = 0.44, P = 0.0067)的分层分析中,T/C基因型的相关性也很明显。然而,在肿瘤位置分层分析中,我们观察到携带C/C基因型的患者在显性和隐性遗传模式下患直肠癌的风险(OR = 7.57, P = 0.0403)高于结肠癌(OR = 4.87, P = 0.234)。总之,我们首次在墨西哥患者中研究了+294T/C (rs2016520)多态性与结直肠癌之间的关系。我们的研究结果表明PPARD的变异可能在结直肠癌的遗传易感性中起重要作用。
{"title":"Protective role of +294 T/C (rs2016520) polymorphism of PPARD in Mexican patients with colorectal cancer.","authors":"M. A. Rosales-Reynoso, L. Wence-Chávez, A. R. Arredondo-Valdez, S. Dumois-Petersen, Patricio Barros-Núñez, M. Gallegos-Arreola, S. Flores-Martínez, José Sánchez-Corona","doi":"10.4238/gmr16019324","DOIUrl":"https://doi.org/10.4238/gmr16019324","url":null,"abstract":"PPARD encodes for peroxisome proliferator-activated receptor delta, which plays a significant role in controlling lipid metabolism, atherosclerosis, inflammation, cancer growth, progression, and apoptosis. Accumulated evidence suggests that the polymorphism rs2016520 in PPARD is associated with lipid metabolism, obesity, metabolic syndrome, and type 2 diabetes mellitus. The aim of this study was to determine whether the single nucleotide polymorphism +294T/C (rs2016520) in PPARD is associated with colorectal cancer (CRC) in the Mexican population. Genomic DNA from 178 CRC patients and 97 healthy blood donors was analyzed. The polymorphism was identified by the polymerase chain reaction-restriction fragment length polymorphism method. Results demonstrated that patients with the T/C genotype for the +294T/C (rs2016520) polymorphism present a protective role against CRC [odds ratio (OR) = 0.39; 95% confidence interval (CI) = 0.22-0.69; P = 0.0008]. This association was also evident for the T/C genotype in the stratified analysis by tumor-node-metastasis stages I+II (OR = 0.26, P = 0.0332) and III+IV (OR = 0.44, P = 0.0067). However, in the stratified analysis by tumor location, we observed an increased risk of rectal cancer (OR = 7.57, P = 0.0403) vs colon cancer (OR = 4.87, P = 0.234) in patients carrying the C/C genotype and under the dominant and recessive models of inheritance. In conclusion, for the first time, the association between the +294T/C (rs2016520) polymorphism and colorectal cancer has been studied in Mexican patients. Our results reveal that variations in PPARD may play a significant role in genetic susceptibility to colorectal cancer.","PeriodicalId":189314,"journal":{"name":"Genetics and molecular research : GMR","volume":"33 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":"128110977","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}
引用次数: 5
Transfer and expression of the rabbit defensin NP-1 gene in lettuce (Lactuca sativa). 兔防御素NP-1基因在莴苣中的转移与表达。
Pub Date : 2017-01-23 DOI: 10.4238/gmr16019333
D. Song, X. Xiong, W. Tu, W. Yao, H. Liang, F. J. Chen, Z. He
Lettuce (Lactuca sativa L.) is an annual plant of the daisy family, Asteraceae, with high food and medicinal value. However, the crop is susceptible to several viruses that are transmitted by aphids and is highly vulnerable to post-harvest diseases, as well as insect and mammal pests and fungal and bacterial diseases. Here, the rabbit defensin gene NP-1 was transferred into lettuce by Agrobacterium-mediated transformation to obtain a broad-spectrum disease-resistant lettuce. Transgenic lettuce plants were selected and regenerated on selective media. The presence of the NP-1 gene in these plants was confirmed by western blot analyses. Resistance tests revealed native defensin NP-1 expression conferred partial resistance to Bacillus subtilis and Pseudomonas aeruginosa, which suggests new possibilities for lettuce disease resistance.
莴苣(lacuca sativa L.)是菊科雏菊科一年生植物,具有很高的食用和药用价值。然而,该作物易受几种由蚜虫传播的病毒的影响,极易受收获后疾病、昆虫和哺乳动物害虫以及真菌和细菌疾病的影响。本研究通过农杆菌介导转化将兔防御素基因NP-1转入莴苣中,获得广谱抗病莴苣。选择转基因生菜植株,在选择培养基上进行再生。western blot分析证实了这些植物中存在NP-1基因。抗性试验表明,天然防御素NP-1表达对枯草芽孢杆菌和铜绿假单胞菌具有部分抗性,这为莴苣抗病提供了新的可能性。
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引用次数: 2
Dynamics of 45S rDNA sites in the cell cycle: fragile sites and chromosomal stability in Lolium and Festuca. 细胞周期中45S rDNA位点的动态:禾草和羊茅的脆性位点和染色体稳定性。
Pub Date : 2017-01-23 DOI: 10.4238/gmr16019156
L. Rocha, G. A. Silva, F. O. Bustamante, R. A. D. Silveira, A. Mittlemann, V. H. Techio
Analyses carried out with fluorescence in situ hybridization (FISH) in C-metaphases of the Lolium-Festuca complex have shown the occurrence of spontaneous fragile sites (FSs) in 45S rDNA regions. FSs are expressed as gaps but they do not result in breaks or chromosomal fragments in these species. These gaps have high DNA condensation observed as thin chromatin fibers that connect the apparent segments of the fragile chromosome, allowing for genomic stability. Assessing the behavior of these regions in the cell cycle of Lolium and Festuca species may lead to a better understanding of the dynamics that preserve stability during cell division. Furthermore, it is interesting to track the dynamics of chromosomes bearing 45S rDNA sites in the cell cycle as well as to observe the expression of FSs with no effect of the mitotic block. We observed variation in both the number and size of 45S FISH signals from the S/G2 phases of interphase and from prophase to anaphase where gaps in 45S rDNA sites also were observed. The change in the degree of condensation of the 45S site begins in the S/G2 phase and appears to be related to the transcriptional demand. Taking into account that the number of 45S rDNA sites tends to be re-established when cells reach telophase, we suggest that the chromatin fiber goes back to the normal condensation level to the anaphase (after segregation), allowing for the approximation of chromosome segments and ensuring dynamics that favor the genomic stability of these species.
荧光原位杂交(FISH)对Lolium-Festuca复合体c -中期进行的分析显示,在45S rDNA区域存在自发脆性位点(FSs)。在这些物种中,FSs表现为间隙,但它们不会导致断裂或染色体片段。这些缝隙具有高度的DNA凝结,就像薄的染色质纤维一样,连接着脆弱的染色体的表面片段,从而保证了基因组的稳定性。评估这些区域在Lolium和Festuca物种细胞周期中的行为可能有助于更好地理解在细胞分裂过程中保持稳定性的动力学。此外,在细胞周期中追踪携带45S rDNA位点的染色体的动态以及观察FSs在没有有丝分裂阻断的情况下的表达是很有趣的。我们观察到45S FISH信号的数量和大小在间期的S/G2期和从前期到后期的变化,45S rDNA位点也有间隙。45S位点缩聚程度的变化始于S/G2期,似乎与转录需求有关。考虑到45S rDNA位点的数量倾向于在细胞到达末期时重新建立,我们建议染色质纤维回到正常的凝聚水平到后期(分离后),允许染色体片段的近似,并确保有利于这些物种基因组稳定性的动力学。
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引用次数: 6
Long non-coding RNA ENST00000457645 reverses cisplatin resistance in CP70 ovarian cancer cells. 长链非编码RNA ENST00000457645逆转CP70卵巢癌细胞的顺铂耐药。
Pub Date : 2017-01-23 DOI: 10.4238/gmr16019411
H. Yan, J. Xia, F. Feng
The objective of this study was to investigate the effect of downregulating long non-coding RNAs (lncRNAs) on the reversal of cisplatin resistance in CP70 ovarian cancer cells, and to identify the underlying mechanism(s) of action. An lncRNA microarray was performed to screen for downregulated lncRNAs in cisplatin-resistant CP70 cells. Expression levels of these lncRNAs were then verified in SKOV3 and SKOV3/DDP cells. Quantitative polymerase chain reaction was conducted to identify the lncRNA most downregulated, which was then synthesized and transfected into CP70 cells. To assess the viability and migration ability of these transfected CP70 cells, methyl thiazolyl tetrazolium and Transwell assays were carried out. In addition, expression levels of apoptosis-related proteins were examined by western blotting. The lncRNA microarray analysis and qPCR identified seven lncRNAs that were significantly downregulated. Transfection of lncRNA ENST00000457645 into CP70 cells markedly inhibited viability and migration ability, and significantly increased expression of apoptotic proteins such as Bax and cleaved caspase-3. lncRNA ENST00000457645 negatively affects the viability and migration of cisplatin-resistant CP70 ovarian cancer cells. The mechanism responsible involves modification of apoptotic protein expression.
本研究的目的是探讨下调长链非编码rna (lncRNAs)在逆转CP70卵巢癌细胞顺铂耐药中的作用,并确定其潜在的作用机制。使用lncRNA芯片筛选顺铂耐药CP70细胞中下调的lncRNA。然后在SKOV3和SKOV3/DDP细胞中验证这些lncrna的表达水平。定量聚合酶链反应鉴定下调幅度最大的lncRNA,合成并转染到CP70细胞中。为了评估这些转染的CP70细胞的活力和迁移能力,进行甲基噻唑四氮唑和Transwell实验。western blotting检测凋亡相关蛋白的表达水平。lncRNA微阵列分析和qPCR鉴定出7个显著下调的lncRNA。转染lncRNA ENST00000457645后,CP70细胞的生存能力和迁移能力明显受到抑制,Bax、cleaved caspase-3等凋亡蛋白的表达明显增加。lncRNA ENST00000457645负性影响顺铂耐药CP70卵巢癌细胞的生存能力和迁移能力。其机制涉及凋亡蛋白表达的修饰。
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引用次数: 26
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Genetics and molecular research : GMR
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