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Variant Analysis of miRNA Regulatory Genes in 35 Sporadic Lung Carcinoma Tumors 35 例散发性肺癌肿瘤中 miRNA 调控基因的变异分析
IF 0.8 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-03-12 DOI: 10.1134/S1607672924600052
Özkan Bağcı, Ebru Marzioğlu Özdemir, Batuhan Şanlıtürk

Lung cancer is one of the cancer types with the highest mortality worldwide. The most frequently mutated genes known to be clinically important in lung cancers are EGFR, BRAF, and KRAS genes. Therefore, the therapeutic agents developed are directed against variants that cause over-activation of the EGFR–KRAS–BRAF–BRAF–MEK/ERK signalling pathway. However, different responses of patients to Tyrosine Kinase Inhibitors (TKIs) suggest that new prognostic biomarkers should be defined and epigenetic mechanisms may be related to this situation. Methods: In this study, sequence analyses of AGO2, DICER, and DROSHA genes involved in miRNA biogenesis and EGFR, KRAS, and BRAF genes were performed in 35 patients with sporadic lung cancer. Results: We found variations in genes involved in miRNA biogenesis that have not been previously reported in the literature. In addition, we found 4 different variants in the EGFR gene that have been described in the literature. In addition, a statistically significant association was found between the presence of mutations in at least one of the genes involved in miRNA biogenesis and metastasis (p:0.02). Conclusions: In conclusion, genomic dysregulation of key miRNA biogenesis genes may be one of the possible reasons for the differential response of patients to therapeutic agents and the development of metastasis in EGFR wild type tumours.

肺癌是全球死亡率最高的癌症类型之一。已知在肺癌临床上最常见的突变基因是表皮生长因子受体(EGFR)、BRAF 和 KRAS 基因。因此,开发的治疗药物都是针对导致表皮生长因子受体-KRAS-BRAF-BRAF-MEK/ERK 信号通路过度激活的变异基因。然而,患者对酪氨酸激酶抑制剂(TKIs)的不同反应表明,应该定义新的预后生物标志物,而表观遗传机制可能与这种情况有关:本研究对35例散发性肺癌患者中参与miRNA生物发生的AGO2、DICER和DROSHA基因以及EGFR、KRAS和BRAF基因进行了序列分析:结果:我们在参与 miRNA 生物发生的基因中发现了以前文献中未报道过的变异。此外,我们还在表皮生长因子受体(EGFR)基因中发现了 4 种不同的变异,这些变异已在文献中有所描述。此外,我们还发现,至少一个参与 miRNA 生物发生的基因出现变异与转移之间存在统计学意义上的显著关联(p:0.02):总之,关键 miRNA 生物发生基因的基因组失调可能是表皮生长因子受体野生型肿瘤患者对治疗药物反应不同和发生转移的原因之一。
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引用次数: 0
Anti-tumor Effect of High Doses of Carbon Ions and X-Rays during Irradiation of Ehrlich Ascites Carcinoma Cells Ex Vivo 大剂量碳离子和 X 射线辐照艾氏腹水癌细胞的体内抗肿瘤效应
IF 0.8 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-03-12 DOI: 10.1134/S1607672924700765
V. E. Balakin, T. A. Belyakova, O. M. Rozanova, E. N. Smirnova, N. S. Strelnikova, E. A. Kuznetsova

The effect of carbon ions (12C) with the energy of 400 MeV/nucleon on the dynamics of induction and growth rate of solid tumors in mice under irradiation of Ehrlich ascites carcinoma cells (EAC) ex vivo at doses of 5–30 Gy relative to the action of equally effective doses of X-ray radiation was studied. The dynamics of tumor induction under the action of 12C and X-rays had a similar character and depended on the dose during 3 months of observation. The value of the latent period, both when irradiating cells with 12C and X-ray, increased with increasing dose, and the interval for tumor induction decreased. The rate of tumor growth after ex vivo irradiation of EAC cells was independent of either dose or type of radiation. The dose at which EAC tumors are not induced within 90 days was 30 Gy for carbon ions and 60 Gy for X-rays. The value of the relative biological effectiveness of carbon ions, calculated from an equally effective dose of 50% probability of tumors, was 2.59.

研究了能量为 400 MeV/nucleon的碳离子(12C)对小鼠体内外照射艾氏腹水癌细胞(EAC)时实体瘤诱导和生长速度的动态影响,照射剂量为 5-30 Gy,相对于同样有效剂量的 X 射线辐射作用。在 3 个月的观察期间,12C 和 X 射线作用下肿瘤诱导的动态具有相似的特征,并取决于剂量。用 12C 和 X 射线照射细胞时,潜伏期的值都随着剂量的增加而增加,肿瘤诱导的间隔时间则缩短。EAC细胞体外照射后的肿瘤生长率与剂量或辐射类型无关。90 天内不诱发 EAC 肿瘤的剂量为碳离子 30 Gy 和 X 射线 60 Gy。根据肿瘤概率为 50%的同等有效剂量计算,碳离子的相对生物有效性值为 2.59。
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引用次数: 0
The Level of Anti-Viral Antigen-Specific Antibodies to EBNA-1 in the Serum of MS Patients Does not Depend on the Severity of the Disease 多发性硬化症患者血清中的 EBNA-1 抗病毒抗原特异性抗体水平与病情严重程度无关
IF 0.8 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-03-12 DOI: 10.1134/S1607672924700753
L. A. Ovchinnikova, S. S. Dzhelad, T. O. Simaniv, M. N. Zakharova,  A. G. Gabibov, Y. A. Lomakin

Multiple sclerosis (MS) is an autoimmune neurodegenerative disease leading to inevitable disability and primarily affecting the young and middle-aged population. Recent studies have shown a direct correlation between the risk of MS development and Epstein–Barr virus (EBV) infection. Analysis of the titer of EBV-specific antibodies among patients with MS and healthy donors among Russian population confirmed that MS is characterized by an increased level of serum IgG binding EBNA-1 (EBV nuclear antigen 1). The number of patients with elevated levels of EBNA-1-specific antibodies does not differ statistically significantly between two groups with diametrically opposite courses of MS: benign MS or highly active MS. It can be assumed that the primary link between EBV and the development of MS is restricted to the initiation of the disease and does not impact its severity.

多发性硬化症(MS)是一种自身免疫性神经退行性疾病,会导致不可避免的残疾,主要影响中青年人群。最近的研究表明,多发性硬化症的发病风险与爱泼斯坦-巴氏病毒(EBV)感染直接相关。对俄罗斯多发性硬化症患者和健康供体的 EBV 特异性抗体滴度分析证实,多发性硬化症的特征是血清中结合 EBNA-1(EBV 核抗原 1)的 IgG 水平升高。EBNA-1特异性抗体水平升高的患者人数在多发性硬化症病程截然相反的两组(良性多发性硬化症或高度活动性多发性硬化症)之间没有明显的统计学差异。可以认为,EB 病毒与多发性硬化症发病之间的主要联系仅限于疾病的起始阶段,而不会影响其严重程度。
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引用次数: 0
Study of the Association of Ouib and Nom with Heterochromatin in Drosophila melanogaster 黑腹果蝇中 Ouib 和 Nom 与异染色质的关联研究
IF 0.8 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-03-12 DOI: 10.1134/S1607672924700741
Y. V. Pekina, V. A. Babosha,  P. G. Georgiev, A. A. Fedotova

In Drosophila, a large group of actively transcribed genes is located in pericentromeric heterochromatin. It is assumed that heterochromatic proteins recruit transcription factors to gene promoters. Two proteins, Ouib and Nom, were previously shown to bind to the promoters of the heterochromatic genes nvd and spok. Interestingly, Ouib and Nom are paralogs of the M1BP protein, which binds to the promoters of euchromatic genes. We have shown that, like M1BP, the Quib and Nom proteins bind to CP190, which is involved in the recruitment of transcription complexes to promoters. Unlike heterochromatic proteins, Ouib and Nom do not interact with the major heterochromatic protein HP1a and bind to euchromatic promoters on polytene chromosomes from the larval salivary glands. The results suggest a new mechanism for the recruitment of transcription factors into the heterochromatic compartment of the nucleus.

在果蝇中,一大批活跃的转录基因位于中心粒周围的异染色质中。据推测,异染色质蛋白将转录因子招募到基因启动子上。之前有研究表明,Ouib 和 Nom 这两种蛋白能与异染色质基因 nvd 和 spok 的启动子结合。有趣的是,Ouib 和 Nom 是 M1BP 蛋白的旁系亲属,而 M1BP 蛋白能与外染色质基因的启动子结合。我们已经证明,与 M1BP 蛋白一样,Quib 和 Nom 蛋白也与 CP190 结合,而 CP190 参与转录复合体与启动子的招募。与异染色质蛋白不同的是,Ouib 和 Nom 不与主要的异染色质蛋白 HP1a 相互作用,而是与幼虫唾液腺多体染色体上的非染色质启动子结合。这些结果表明,转录因子被招募到细胞核的异染色区是一种新的机制。
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引用次数: 0
Experimental and Model Study of a Swirling Fluid Flow in a Converging Channel As a Simulation of Blood Flow in the Heart and Aorta 作为心脏和主动脉血流模拟的汇流槽中漩涡流的实验和模型研究
IF 0.8 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-03-12 DOI: 10.1134/S1607672924700777
Y. E. Zharkov, S. T. Zhorzholiani, A. A. Sergeev, A. V. Agafonov, A. Y. Gorodkov,  L. A. Bockeria

Study of swirling flows in channels corresponding to the static approximation of flow channels of the heart and major vessels with a longitudinal–radial profile zR2 = const and a concave streamlined surface at the beginning of the longitudinal coordinate has been carried out. A comparative analysis of the flow structure in channel configurations zRN = const, where N = –1, 1, 2, 3, in the absence and presence of a concave surface was carried out. The numerical modeling was compared with the results of hydrodynamic experiments on the flow characteristics and the shape of the flow lines. The numerical model was used to determine the velocity structure, viscous friction losses, and shear stresses. Numerical modeling of steady-state flows for channels without a concave surface showed that in the channel zR2 = const there is a stable vortex flow structure with the lowest viscous friction losses. The presence of a concave surface of sufficient size significantly reduces viscous friction losses and shear stresses in both the steady state and pulsed modes.

研究了与心脏和大血管流道静态近似的流道中的漩涡流,该流道具有纵向-径向剖面 zR2 = const 和纵向坐标起点的凹流线型表面。对通道配置 zRN = const(其中 N =-1、1、2、3)中没有凹面和有凹面时的流动结构进行了比较分析。数值模型与流体力学实验结果在流动特性和流线形状方面进行了比较。数值模型用于确定速度结构、粘性摩擦损失和剪应力。对无凹面水道的稳态流进行的数值建模表明,在 zR2 = const 的水道中,存在稳定的涡流结构,粘性摩擦损失最小。在稳态和脉冲模式下,足够大的凹面都能显著降低粘性摩擦损失和剪切应力。
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引用次数: 0
PRPF19 mRNA Encodes a Small Open Reading Frame That Is Important for Viability of Human Cells PRPF19 mRNA 编码一个小型开放阅读框,对人类细胞的存活率非常重要。
IF 0.8 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-03-12 DOI: 10.1134/S1607672923700722
N. M. Shepelev, A. O. Kurochkina,  O. A. Dontsova, M. P. Rubtsova

High-throughput ribosome profiling demonstrates the translation of thousands of small open reading frames located in the 5′ untranslated regions of messenger RNAs (upstream ORFs). Upstream ORF can both perform a regulatory function by influencing the translation of the downstream main ORF and encode a small functional protein or microprotein. In this work, we showed that the 5′ untranslated region of the PRPF19 mRNA encodes an upstream ORF that is translated in human cells. Inactivation of this upstream ORF reduces the viability of human cells.

高通量核糖体分析显示,位于信使 RNA(上游 ORF)5'非翻译区的数千个小型开放阅读框在进行翻译。上游 ORF 既能通过影响下游主 ORF 的翻译发挥调控功能,也能编码小型功能蛋白或微量蛋白。在这项工作中,我们发现 PRPF19 mRNA 的 5' 非翻译区编码一个上游 ORF,该 ORF 在人体细胞中被翻译。该上游 ORF 失活会降低人体细胞的活力。
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引用次数: 0
Regulation of Intestinal Flora and Immune Response by Cyanidin Exhibits Protective Effect against Type-2 Diabetes in Rat Model 矢车菊素调节肠道菌群和免疫反应对大鼠模型中的 2 型糖尿病有保护作用
IF 0.8 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-02-20 DOI: 10.1134/S1607672923600422
Qingyan Lu, Qiannan Yan, Xiaojie Li

In the current study the effects of metformin and cyanidin on the immune system and intestinal flora in rats with type-2 diabetes was investigated. The findings showed that metformin or cyanidin treatment considerably reduced the rise in body weight and glucose levels induced by type-2 diabetes. The type-2 diabetic rats’ glucose tolerance was significantly increased by cyanidin administration comparable to that of metformin. Cyanidin administration resulted in a significant reduction in serum cholesterol and low-density lipoprotein (LDL) levels in rats with type-2 diabetes. Treatment with cyanidin significantly increased the ratio of high-density lipoprotein to low-density lipoprotein in type-2 diabetes rats. Cyanidin administration significantly raised the ratio of Firmicutes to Bacteroidetes in the fecal samples of type-2 diabetic rats compared to the model group. In comparison to the model group, it also significantly raised the levels of Lactobacillus intestinalis, Lactobacillus gasseri, and Lactobacillus reuteri in the type-2 diabetes rats. In type-2 diabetes rat fecal samples, the abundance of Christensenellaceae significantly increased while Enterobacteriaceae and Proteobacteria were found to decrease upon cyanidin administration. Furthermore, cyanidin administration to the rats with type-2 diabetes significantly improved the glucose homeostasis. In conclusion, the study demonstrates that cyanidin enhances glucose homeostasis in rats with type-2 diabetes, potentially through controlling intestinal flora. Thus, cyanidin may be looked into more as a possible therapeutic agent for type 2 diabetes.

本研究调查了二甲双胍和青花素对 2 型糖尿病大鼠免疫系统和肠道菌群的影响。研究结果表明,二甲双胍或青花素能显著降低 2 型糖尿病大鼠体重和血糖水平的升高。与二甲双胍相比,服用青花素能显著提高 2 型糖尿病大鼠的葡萄糖耐量。服用青花素可显著降低 2 型糖尿病大鼠的血清胆固醇和低密度脂蛋白(LDL)水平。青花素能显著提高 2 型糖尿病大鼠体内高密度脂蛋白与低密度脂蛋白的比率。与模型组相比,青花素能明显提高2型糖尿病大鼠粪便样本中的固缩菌与类杆菌的比例。与模型组相比,青花素还能显著提高 2 型糖尿病大鼠肠道乳杆菌、加塞利乳杆菌和吕特氏乳杆菌的含量。在 2 型糖尿病大鼠的粪便样本中,发现在施用青花素后,Christensenellaceae 的丰度显著增加,而 Enterobacteriaceae 和 Proteobacteria 则减少。此外,给 2 型糖尿病大鼠服用青花素还能明显改善其血糖稳态。总之,该研究表明,青花素可通过控制肠道菌群改善 2 型糖尿病大鼠的葡萄糖稳态。因此,可将青花素作为一种治疗 2 型糖尿病的药物进行更多研究。
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引用次数: 0
Quantitative Description of the N-Protein of the SARS-CoV-2 Virus Degradation in Cells Stably Expressing It under the Influence of New Modular Nanotransporters 新型模块化纳米转运体影响下稳定表达 SARS-CoV-2 病毒的细胞中 N 蛋白降解的定量描述
IF 0.8 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-02-20 DOI: 10.1134/S1607672923700709
Y. V. Khramtsov, A. V. Ulasov, T. N. Lupanova,  G. P. Georgiev,  A. S. Sobolev

Two eukaryotic cell lines, A549 and A431, with stable expression of the nucleocapsid protein (N-protein) of the SARS-CoV-2 virus fused with the red fluorescent protein mRuby3 were obtained. Using microscopy, the volumes of the cytoplasm and nucleus were determined for these cells. Using quantitative immunoblotting techniques, the concentrations of the N-mRuby3 fusion protein in their cytoplasm were assessed. They were 19 and 9 μM for A549 and A431 cells, respectively. Using these concentrations, the initial rate of N-protein degradation in the studied cells was estimated from the decrease in cell fluorescence. In A549 and A431 cells, it was the same (84 nM per hour). The approach of quantitatively describing the degradation process can be applied to analyze the effectiveness of a wide class of antiviral drugs that cause degradation of viral proteins.

研究人员获得了稳定表达融合了红色荧光蛋白 mRuby3 的 SARS-CoV-2 病毒核壳蛋白(N 蛋白)的两种真核细胞系 A549 和 A431。利用显微镜测定了这些细胞的细胞质和细胞核的体积。利用定量免疫印迹技术,评估了细胞质中 N-mRuby3 融合蛋白的浓度。A549 和 A431 细胞的浓度分别为 19 和 9 μM。利用这些浓度,可以根据细胞荧光的下降估算出研究细胞中 N 蛋白的初始降解率。在 A549 和 A431 细胞中,降解速度相同(每小时 84 nM)。定量描述降解过程的方法可用于分析导致病毒蛋白降解的多种抗病毒药物的有效性。
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引用次数: 0
Modular Nanotransporters Capable of Causing Intracellular Degradation of the N-Protein of the SARS-CoV-2 Virus in A549 Cells with Temporary Expression of This Protein Fused with a Fluorescent Protein mRuby3 在 A549 细胞中临时表达融合了荧光蛋白 mRuby3 的 SARS-CoV-2 病毒 N 蛋白的模块化纳米转运体可引起该蛋白的胞内降解
IF 0.8 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-02-20 DOI: 10.1134/S1607672923700710
Y. V. Khramtsov, A. V. Ulasov, T. N. Lupanova,  G. P. Georgiev,  A. S. Sobolev

Modular nanotransporters (MNTs) containing an antibody-like molecule, monobody, to the N‑protein of the SARS-CoV-2 virus, as well as an amino acid sequence that recruits the Keap1 E3 ligase (E3BP) were created. This MNT also included a site for cleavage of the E3BP monobody from the MNT in acidic endocytic compartments. It was shown that this cleavage by the endosomal protease cathepsin B leads to a 2.7-fold increase in the affinity of the E3BP monobody for the N-protein. Using A549 cells with transient expression of the N-protein fused with the fluorescent protein mRuby3, it was shown that incubation with MNT leads to a significant decrease in mRuby3 fluorescence. It is assumed that the developed MNTs can serve as a basis for the creation of new antiviral drugs against the SARS-CoV-2 virus.

我们创建了模块化纳米转运体(MNTs),其中含有一种类似于抗体的分子(单体),可与 SARS-CoV-2 病毒的 N 蛋白结合,还含有一种可诱导 Keap1 E3 连接酶(E3BP)的氨基酸序列。这种 MNT 还包括一个用于在酸性内细胞区室中将 E3BP 单体从 MNT 上裂解的位点。研究表明,内泌体蛋白酶 cathepsin B 的这种裂解会使 E3BP 单体对 N 蛋白的亲和力增加 2.7 倍。使用瞬时表达与荧光蛋白 mRuby3 融合的 N 蛋白的 A549 细胞,结果表明与 MNT 一起孵育会导致 mRuby3 荧光显著降低。据推测,所开发的 MNT 可作为开发针对 SARS-CoV-2 病毒的新型抗病毒药物的基础。
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引用次数: 0
Effects of Chromatin Structure Modifiers on the trans-Acting Heterochromatin Position Effect in Drosophila melanogaster 染色质结构修饰剂对黑腹果蝇反式异染色质位置效应的影响
IF 0.8 4区 生物学 Q4 BIOCHEMISTRY & MOLECULAR BIOLOGY Pub Date : 2024-02-20 DOI: 10.1134/S160767292470073X
A. A. Solodovnikov, S. A. Lavrov, A. S. Shatskikh,  V. A. Gvozdev

The heterochromatin position effect is manifested in the inactivation of euchromatin genes transferred to heterochromatin. In chromosomal rearrangements, genes located near the new eu-heterochromatin boundary in the rearrangement (cis-inactivation) and, in rare cases, genes of a region of the normal chromosome homologous to the region of the eu-heterochromatin boundary of the chromosome with the rearrangement (trans-inactivation) are subject to inactivation. The In(2)A4 inversion is able to trans-inactivate the UAS-eGFP reporter gene located on the normal chromosome. We knockdown a number of chromatin proteins using temperature-controlled RNA interference and investigated the effect of knockdown on trans-inactivation of the reporter. We found suppression of trans-inactivation by knockdowns of Su(var)2-HP2, a protein that binds to the key heterochromatin protein HP1a, SAYP, a subunit of the chromatin remodelling complex, and Eggless histone methyltransferase (SETDB1), which introduces a H3K9me3 histone mark, recognized by the HP1a protein. The method of studying the effects of gene knockdown on heterochromatin position effects presented in this work is of independent methodological interest.

异染色质位置效应表现为转移到异染色质的优染色质基因失活。在染色体重排中,位于重排中新的优染色质-异染色质边界附近的基因(顺式失活),在极少数情况下,正常染色体上与重排染色体的优染色质-异染色质边界同源区域的基因(反式失活)也会失活。In(2)A4 反转能反式灭活位于正常染色体上的 UAS-eGFP 报告基因。我们利用温控 RNA 干扰敲除了一些染色质蛋白,并研究了敲除对报告基因反式失活的影响。我们发现,敲除与关键异染色质蛋白HP1a结合的蛋白Su(var)2-HP2、染色质重塑复合物的亚基SAYP和引入HP1a蛋白识别的H3K9me3组蛋白标记的Eggless组蛋白甲基转移酶(SETDB1)会抑制反式失活。这项工作提出的研究基因敲除对异染色质位置效应影响的方法具有独立的方法学意义。
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引用次数: 0
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