Biochemical Genetics最新文献

Alzheimer's disease (AD) and mild cognitive impairment (MCI) are a serious global public health problem. The aim of this study was to analyze the key molecular pathological mechanisms that occur in early AD progression as well as MCI. Expression profiling data from brain homogenates of 8 normal volunteers, and 6 patients with prodromal AD who had developed MCI were analyzed, and the data were obtained from GSE12685. Further, overexpression of GABBR2 was achieved in human neuroblastoma cell lines SH-SY5Y and BE(2)-M17 using expression plasmid transfection. GABBR2 was significantly overexpressed in brain tissues of patients with prodromal AD who had developed MCI, as compared to normal brains. Moreover, GABBR2 overexpressing cells showed a significant increase in intracellular Ca²⁺ concentration, a large amount of reactive oxygen species production, a large opening of the mitochondrial permeability transition pore and a significant increase in apoptosis compared with control cells. GABBR2 overexpression was significantly involved in early AD progression and MCI by causing cellular events such as intracellular Ca²⁺ imbalance, oxidative stress, and mitochondrial dysfunction.

Pulmonary hypertension (PH) is a progressive disease characterized by vascular reHypoxiaing, endothelial cell dysfunction, and inflammation. Liver Kinase B1 (LKB1, also known as STK11) is a central regulator of cell polarity and energy homeostasis. However, its specific role and mechanism of action in PH remain unclear. Human pulmonary arterial endothelial cells (hPAECs) were cultured in vitro to establish PH cell Hypoxias under normoxic and hypoxic conditions. The expression of LKB1 was detected by reverse transcriptase-quantitative polymerase chain reaction (RT-qPCR) and western blotting, and its effect on hPAECs function was investigated by overexpression and inhibition of LKB1. Furthermore, cell proliferation was assessed using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, apoptosis was measured by flow cytometry, inflammatory cytokine secretion was evaluated using enzyme-linked immunosorbent assay (ELISA), and the expression of AMP-activated protein kinase (AMPK) signaling pathway-related proteins was analyzed by western blotting. LKB1 expression was significantly reduced in hypoxia-treated hPAECs compared with that in normoxic controls, and LKB1 overexpression significantly ameliorated the hypoxia-induced decrease in cell proliferation and increase in apoptosis as well as inflammatory factor secretion. The AMPK agonist (GSK621) reversed the dysfunction caused by LKB1 inhibition, indicating that LKB1 regulates hPAECs function through the AMPK signaling pathway. LKB1 plays a protective role in PH by inhibiting hPAECs dysfunction via activation of the AMPK pathway.

The long non-coding RNA Growth Arrest-Specific 5 (GAS5) is pivotal in modulating key signaling pathways by functioning as a molecular sponge for microRNAs (miRNAs). GAS5 is notably recognized for its antitumor properties, primarily through its ability to sequester oncogenic miRNAs, thereby influencing critical pathways such as p53, Wnt/β-catenin, and PI3K/Akt, all of which are integral to cell proliferation, apoptosis, and metastasis. The disruption of GAS5-miRNA interactions has been implicated in various malignancies, reinforcing its potential as both a biomarker and a therapeutic target. This paper delves into the intricate signaling cascades affected by GAS5-miRNA interactions and thoroughly investigates the diagnosis and treatment prospects associated with GAS5. Moreover, it addresses both the challenges and opportunities for translational applicability of these findings in clinical environments. The study emphasizes GAS5's significance within the cancer molecular landscape and posits that precise modulation of GAS5-miRNA interactions could catalyze transformative developments in cancer diagnostics and therapeutic approaches. This comprehensive review not only highlights the critical role of non-coding RNAs in cancer biology but also aims to lay the groundwork for future investigations aimed at harnessing these insights for therapeutic interventions.

Retinoblastoma (RB) is an aggressive form of eye cancer. β-Asarone is a bioactive component isolated from the medicinal plant Acorus tatarinowii Schott and has anticancer effects on various human cancers. However, reports regarding the role of β-Asarone in RB remain limited. Our study investigates the mechanisms of β-Asarone in regulating drug resistance in RB, providing a theoretical foundation for RB treatment. A carboplatin-resistant RB cell line was established and treated with β-Asarone, followed by overexpression of long non-coding RNA (lncRNA) urothelial carcinoma-associated 1 (UCA1). The half-maximal inhibitory concentration and cell apoptosis were determined. The levels of lncRNA UCA1/miR-206/neuropilin 1 (NRP1) were measured. The subcellular localization of lncRNA UCA1 was examined. The binding relationships between lncRNA UCA1 and microRNA (miR)-206, and between miR-206 and NRP1 were analyzed. NRP1 expression was analyzed by Western blot assay. We found that β-Asarone downregulated lncRNA UCA1 expression in carboplatin-resistant RB cells. Overexpression of lncRNA UCA1 reversed the inhibitory effect of β-Asarone on cell drug resistance and cell proliferation and reduced apoptosis. LncRNA UCA1 functioned as a sponge for miR-206, which suppressed NRP1 expression. Inhibition of miR-206 or overexpression of NRP1 could partially reverse the suppressive effect of β-Asarone on RB cell drug resistance. In conclusion, β-Asarone suppresses RB cell drug resistance through the lncRNA UCA1/miR-206/NRP1 axis.

Pearl millet {Pennisetum glaucum (L.) R. Br} is a C4 panicoid cereal millet crop grown in arid and semi-arid regions in Africa and Asia for food and fodder. This study involves the evaluation of the genetic diversity of 28 worldwide germplasm collection of pearl millet by genetic markers polymorphism and drought tolerance indices. The genetic diversity was expressed by 51 alleles of 9 ISSR markers that showed 96.43% total polymorphism and 11.76 alleles per marker. Cluster analysis of ISSR markers polymorphism divided the 28 genotypes into four clusters partially in agreement with their origin. The application of drought stress simulated by 20% PEG₆₀₀₀ treatment, retarded the germination percentage, and reduced shoot and root length, seedling fresh and dry weights. Drought tolerance indices (DTIs) were calculated based on the response of the seedling traits under drought stress compared to the control seedlings. ANOVA revealed statistically significant variation among the genotypes (P ≤ 0.05), except for seedling fresh weight (P = 0.17 > 0.05) under control conditions and seedling dry weight (P = 0.99 > 0.05) under drought conditions. Genotypes having higher DTIs for three traits are regarded drought resistant, i.e., those from India, Ethiopia, Pakistan, and Nigeria. The calculated heritability values indicated that seedlings dry weight is the least trait affected by drought stress whereas root length is the most influenced trait. Hierarchical clustering, based on the DTI values, also grouped the genotypes partially concomitant to their origin. The correlation analysis demonstrated a modest positive correlation between shoot length and root length. A low correlation of r ≤ 0.12 was observed between the morphological DTI matrix and the genetic matrix. Nevertheless, high levels of genetic diversity were identified among the examined genotypes that may face genetic erosion by climatic constraints, and a high potential for creating agronomically superior cultivars by crossing widely divergent genotypes.

Mycobacterium tuberculosis (Mtb) remains a leading infectious disease responsible for millions of deaths. RNA sequencing is a rapidly growing technique and a powerful approach to understanding host and pathogen cross-talks via transcriptional responses. However, its application is limited due to the high costs involved.This study is a preliminary attempt to understand host-pathogen cross-talk during TB infection in different TB clinical cohorts using two biological fluids: Whole blood and serum exosomes (EXO). We conducted an RNA-sequencing machine-learning approach using 20 active TB (ATB), 11 latent TB (LTB), three healthy control (HC) whole blood datasets, and two ATB, LTB, and HC serum EXO datasets. During the study, host-derived differentially expressed genes (DEGs) were identified in both whole blood and EXOs, while EXOs were successful in identifying pathogen-derived DEGs only in LTB. The majority of the DEGs in whole blood were up-regulated between ATB and HC, and ATB and LTB, while down-regulated between LTB and HC, which was vice versa for the EXOs, indicating different mechanisms in response to different states of TB infection across the two different biological samples. The pathway analysis revealed that whole blood gene signatures were mainly involved in host immune responses, whereas exosomal gene signatures were involved in manipulating the host's cellular responses and supporting Mtb survival. Overall, identifying both host and pathogen-derived gene signatures in different biological samples for intracellular pathogens like Mtb is vital to decipher the complex interplay between the host and the pathogen, ultimately leading to more successful future interventions.

Hypothyroidism is the most prevalent thyroid disorder and leads to adverse effects on the human body. Serum thyroid stimulating hormone (TSH) values have been related to polymorphisms in multiple genes that may be involved in the regulation of thyroid function. The single nucleotide polymorphism (SNP) rs2046045 is situated in the intron region of the phosphodiesterase 8B (PDE8B) gene, which encodes a high-affinity cyclic adenosine monophosphate (cAMP)-specific phosphodiesterase widely expressed in thyroid tissue. The principal goal of the present study was to investigate the association between the SNP rs2046045 of the PDE8B gene and hypothyroidism. The study was designed as a case-control study, and a total of 160 hypothyroid and 160 healthy controls were involved. Blood samples were drawn from each individual, and deoxyribonuleic acid (DNA) was separated with a suitable DNA isolation kit. For genotyping, the polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) technique was employed. The IBM Statistical Package for Social Sciences (SPSS) 25.0 was utilized to analyze the statistical data. Age differences between the patients and controls were not observed in the present study. The genotype frequency of homozygous wild type (TT), homozygous mutate type (GG), and heterozygous (GT) was 45%, 2.5%, and 52.5%, respectively, in control subjects and 27.5%, 11.25%, and 61.25%, respectively, in cases, and showed a significant difference (p = 0.0002). The minor G allele frequency is elevated in hypothyroid patients as compared to healthy control subjects (41.87% vs. 28.75%), p = 0.0005. The presence of the mutant allele G of rs2046045 in the PDE8B gene correlates with elevated serum TSH levels in hypothyroid patients.

Colorectal cancer (CRC) is a common malignancy that claims the life of many patients. Nucleolar RNA host gene 16 (SNHG16) has been identified as an oncogene in CRC development. However, the role and mechanism of SNHG16 in CRC remain unclear. A total of 27 cases of CRC tumor tissues and adjacent tissues were collected to investigate the expression and correlation among SNHG16, miR-324-3p, ELK4 and PD-L1 using qRT-PCR, western blot and Pearson analysis. Cell proliferation, migration and invasion abilities were determined using CCK-8 and transwell assays. The cytotoxicity of CD8 + T cells and the apoptosis of CD8⁺ T cells was evaluated by LDH assay and flow cytometry, respectively. Dual luciferase assay, RIP and ChIP methods were performed to verify molecular interactions. Our results showed that SNHG16, ELK4 and PD-L1 expression were abnormally elevated and miR-324-3p expression was decreased in tumor tissues from CRC patients and CRC cells. SNHG16 silencing resulted in suppression of cell growth, metastasis, and immune escape of CRC cells, which was reversed by miR-324-3p inhibitor and ELK4 overexpression. Mechanistically, SNHG16 acted as a competitive endogenous RNA to enhance ELK4 expression by sponging miR-324-3p, thereby provoking the transcription of PD-L1. Our results demonstrated that SNHG16 silencing led to the suppression of cell growth, metastasis, and immune escape of CRC cells through mediating miR-324-3p/ELK4/PD-L1 axis, offering promising targets for CRC treatment.

Cervical cancer is one of the most common cancers worldwide. Many studies have reported the involvement of various miRNAs in cervical cancer progression. Our study was centered at investigating how let-7c-5p affected cervical cancer migration and invasion by regulating cell adhesion and its molecular mechanism. Bioinformatics was used for the analysis on differentially expressed mRNAs in cervical cancer and the prediction of their upstream regulatory miRNAs. Immunohistochemistry was performed to assess the expression of CHD7 in cervical cancer tissue. qRT-PCR was performed for examining how much let-7c-5p and CHD7 were expressed. Dual-luciferase assay was performed to verify the regulatory relationship between CHD7 and let-7c-5p. The CCK-8 and transwell assays helped in detecting cell viability, invasion and migration. The ability by which cells adhered to each other was detected by employing cell adhesion assay. In addition, the expression levels of the proteins related to cell adhesion and CHD7 were detected by Western blot. A remarkable high expression-level of CHD7 was discovered in cervical cancer tissues and cells. The cell viability, migration and invasiveness could be suppressed by the knockdown of CHD7 which could also attenuate the expression of cell adhesion-related proteins. Bioinformatics analysis showed that CHD7 had an upstream regulatory gene, miRNA-let-7c-5p, which was markedly lowly expressed in cervical cancer tissues and cells. To validate the binding relationship between CHD7 and let-7c-5p, dual-luciferase assay was performed. Rescue experiments revealed that the cancer-inhibiting effect of let-7c-5p in cervical cancer could be reversed by overexpressed CHD7. let-7c-5p regulates cell adhesion and attenuates cervical cancer migration and invasiveness by targeting CHD7. It indicates that the involvement of let-7c-5p/CHD7 axis is of significance in cervical cancer progression, which opens up new possibilities for us to develop novel clinical treatments for cervical cancer.

Deubiquitinases play essential roles in hepatocellular carcinoma (HCC) progression, however, the role of ubiquitin-specific peptidase 21 (USP21) in HCC development remains unclear. The present work aims to analyze the effect of USP21 on tumor property of HCC cells and the underlying mechanism. mRNA expression levels of USP21 and H2BFS were analyzed by quantitative real-time polymerase chain reaction. Protein expression of USP21, E-cadherin, N-cadherin, Vimentin, H2BFS and methyltransferase 3 (METTL3) was assessed by western blotting assay or immunohistochemistry assay. Clonogenicity assay was used to analyze cell proliferation. Flow cytometry assay was performed to quantify apoptotic rate of cells. Wound-healing assay and transwell assay were conducted to analyze cell migration and invasion, respectively. Xenograft mouse model assay was performed to determine the effect of USP21 knockdown on tumor formation. m6A RNA immunoprecipitation assay (MeRIP) was used to analyze the effect of METTL3 silencing on methylated level of USP21. USP21 expression was upregulated in HCC tissues and cells when compared with control groups. USP21 silencing inhibited proliferation, migration and invasion and induced apoptosis of HCC cells, accompanied by the increased E-cadherin protein expression and decreased N-cadherin and Vimentin protein expression. Moreover, USP21 knockdown delayed tumor formation in vivo. USP21 stabilized H2BFS by deubiquitination, and H2BFS overexpression attenuated USP21 silencing-induced effects in HCC cells. Further, METTL3-mediated m6A methylation of USP21. METTL3-mediated m6A methylation of USP21 promoted HCC progression by stabilizing H2BFS through deubiquitination.