MicroRNA-520c-3p Modulates Doxorubicin-Chemosensitivity in HepG2 Cells

2020 ◽  
Vol 21 (2) ◽  
pp. 237-245 ◽  
Author(s):  
Mohamed A. Ragheb ◽  
Marwa H. Soliman ◽  
Emad M. Elzayat ◽  
Mervat S. Mohamed ◽  
Nada El-Ekiaby ◽  
...  
Keyword(s):  
Hepg2 Cells ◽  
Therapeutic Strategy ◽  
Suppressive Effect ◽  
Expression Level ◽  
Tumor Suppressor Function ◽  
Protein Expression Level ◽  
Blot Technique ◽  
Induction Of Apoptosis ◽  
The Impact

Background: Doxorubicin (DOX) is the most common drugs used in cancer therapy, including Hepatocellular Carcinoma (HCC). Drug resistance, is one of chemotherapy’s significant problems. Emerging studies have shown that microRNAs (miRNAs) could participate in regulating this mechanism. Nevertheless, the impact of miRNAs on HCC chemoresistance is still enigmatic. Objective: Investigating the role of miR-520c-3p in enhancement of anti-tumor effect of DOX against HepG2 cells. Methods: Expression profile for liver related miRNAs (384 miRNAs) has been analyzed on HepG2 cells treated with DOX using qRT-PCR. miR-520c-3p, the most deregulated miRNA, was selected for combination treatment with DOX. Expression level for LEF1, CDK2, CDH1, VIM, Mcl-1 and TP53 was evaluated in miR-520c-3p transfected cells. Cell viability, colony formation, wound healing as well as apoptosis assays have been demonstrated. Furthermore, Mcl-1 protein level was measured using western blot technique. Results: The present data indicated that miR-520c-3p overexpression could render HepG2 cells chemo-sensitive to DOX through enhancing its suppressive effects on proliferation, migration, and induction of apoptosis. The suppressive effect of miR-520c-3p involved altering the expression levels of some key regulators of cell cycle, proliferation, migration and apoptosis including LEF1, CDK2, CDH1, VIM, Mcl-1 and TP53. Interestingly, Mcl-1 was found to be one of the potential targets of miR-520c-3p, and its protein expression level was down-regulated upon miR-520c-3p overexpression. Conclusion: Our data referred to the tumor suppressor function of miR-520c-3p that could modulate chemosensitivity of HepG2 cells toward DOX treatment, providing a promising therapeutic strategy in HCC.

scholarly journals Effect of Inflammation on Female Gonadotropin-Releasing Hormone (GnRH) Neurons: Mechanisms and Consequences

2020 ◽  
Vol 21 (2) ◽  
pp. 529 ◽  
Author(s):  
Klaudia Barabás ◽  
Edina Szabó-Meleg ◽  
István M. Ábrahám
Keyword(s):  
Suppressive Effect ◽  
Gonadotropin Releasing Hormone ◽  
Releasing Hormone ◽  
Inflammatory Signals ◽  
Cholinergic Pathway ◽  
Gnrh Neurons ◽  
Functional Consequences ◽  
Anti Inflammatory ◽  
The Impact

Inflammation has a well-known suppressive effect on fertility. The function of gonadotropin-releasing hormone (GnRH) neurons, the central regulator of fertility is substantially altered during inflammation in females. In our review we discuss the latest results on how the function of GnRH neurons is modified by inflammation in females. We first address the various effects of inflammation on GnRH neurons and their functional consequences. Second, we survey the possible mechanisms underlying the inflammation-induced actions on GnRH neurons. The role of several factors will be discerned in transmitting inflammatory signals to the GnRH neurons: cytokines, kisspeptin, RFamide-related peptides, estradiol and the anti-inflammatory cholinergic pathway. Since aging and obesity are both characterized by reproductive decline our review also focuses on the mechanisms and pathophysiological consequences of the impact of inflammation on GnRH neurons in aging and obesity.

scholarly journals Study of Using Thymbra spicata leaves to reduce the toxic immunosuppresive effect of aflatoxin in broilers

2008 ◽  
Vol 32 (1) ◽  
pp. 140-147
Author(s):  
Hatem M. M. Al-Naemey
Keyword(s):  
Immune Response ◽  
Newcastle Disease ◽  
Suppressive Effect ◽  
The Third ◽  
The Impact ◽  
Thymbra Spicata ◽  
Newcastle Disease Vaccine ◽  
Protection Ratio

The present study performed to evaluate the impact of addition ofThymbra spicata leaves powder of aflatoxin contaminated ration inreduction of the aflatoxin immuno suppressive effect in the immuneresponse induced by Newcastle disease vaccine in broilers.The study conducted in three groups, The first group fed aflatoxin freeration, second group fed aflatoxin contaminated ration without anytreatment, while the third group fed aflatoxin contaminated rationsupplemented of 2% Thymbra spicata leaves powder.All chick groups were vaccinated twice with Newcastle disease vaccine(NDV) at 10 and 20 days of age. ELIZA test was carried out to estimate thehumeral immune response at 6, 19 and 31 days of age. Infection challengwere done at day 32 of age with velogenic ND strain (Z-2003).The results showed that the first group has high significante antibodies titerreach to 3239±249.3 at 19days of age and 10186+845.3 at 31 days of agewith significant level 0.01 and high protection ratio against challenge(100%)at 32 days of age, while the second group has the low antibodies titerreach to 2622.6±27.3 at 19 days of age and 5712.5±786.9at 31daysof ageand low protection ratio against challenge(66.67), the third group’santibodies titer and protection ratio lower than first group and higher thansecond group ,the titers are 3060±542.5 at 19 days of age and 5712.5±786.9at31 days of age while the protection ratio against challenge is 80% at 32days of age .The results showed the immuno suppressive effect of aflatoxin as in secondgroup and the role of Thymbra spicata leaves powder in reduction of theseeffect.

scholarly journals Viral cGAMP nuclease reveals the essential role of DNA sensing in protection against acute lethal virus infection

2020 ◽  
Vol 6 (38) ◽  
pp. eabb4565
Author(s):  
Bruno Hernáez ◽  
Graciela Alonso ◽  
Iliana Georgana ◽  
Misbah El-Jesr ◽  
Rocío Martín ◽  
...  
Keyword(s):  
Virus Infection ◽  
Immune Responses ◽  
Therapeutic Strategy ◽  
Innate Immune ◽  
Protective Mechanism ◽  
Innate Immune Responses ◽  
Dna Sensing ◽  
Nuclease Domain ◽  
The Impact

Cells contain numerous immune sensors to detect virus infection. The cyclic GMP-AMP (cGAMP) synthase (cGAS) recognizes cytosolic DNA and activates innate immune responses via stimulator of interferon genes (STING), but the impact of DNA sensing pathways on host protective responses has not been fully defined. We demonstrate that cGAS/STING activation is required to resist lethal poxvirus infection. We identified viral Schlafen (vSlfn) as the main STING inhibitor, and ectromelia virus was severely attenuated in the absence of vSlfn. Both vSlfn-mediated virulence and STING inhibitory activity were mapped to the recently discovered poxin cGAMP nuclease domain. Animals were protected from subcutaneous, respiratory, and intravenous infection in the absence of vSlfn, and interferon was the main antiviral protective mechanism controlled by the DNA sensing pathway. Our findings support the idea that manipulation of DNA sensing is an efficient therapeutic strategy in diseases triggered by viral infection or tissue damage–mediated release of self-DNA.

scholarly journals New insights into the role of PTCH1 protein in serous ovarian carcinomas

2021 ◽  
Author(s):  
Valentina Karin-Kujundzic ◽  
Adriana Covarrubias-Pinto ◽  
Anita Skrtic ◽  
Semir Vranic ◽  
Ljiljana Serman
Keyword(s):  
Ovarian Cancer ◽  
Protein Expression ◽  
Cell Lines ◽  
Expression Level ◽  
Tumor Promoter ◽  
Protein Expression Level ◽  
Ovarian Carcinomas ◽  
Tissue Samples ◽  
Serous Ovarian Carcinoma

Abstract Background The Hedgehog (Hh) signaling pathway is essential for normal embryonic development, while its hyperactivation in adult organism is associated with development of various cancers, including ovarian cancer. The role of the Hh signaling pathway in ovarian cancer, as well as in certain histological subtypes of ovarian cancer, is poorly understood. Therefore, we investigated the role of PTCH1 protein and changes in the promoter methylation status of the corresponding gene, in a cohort of low- (LGSC) and high-grade (HGSC) serous ovarian carcinomas and HGSC cell lines (OVCAR5, OVCAR8 and OVSAHO). Methods PTCH1 protein expression level was analyzed using immunohistochemistry in tissue samples, and by immunofluorescence and Western blot in cell lines. DNA methylation pattern of PTCH1 gene were analyzed by methylation-specific PCR (MSP). Mann-Whitney U test was used to compare differences in expression of PTCH1 protein among ovarian tumor samples compared with normal tissue samples, while Spearman’s correlation was used to test the association between DNA promoter methylation of the PTCH1 gene and expression of the corresponding protein. Results PTCH1 protein expression level was significantly higher in HGSCs and LGSCs compared with control tissues (healthy ovaries and fallopian tubes). Similarly, cancer cell lines exhibited significantly higher PTCH1 protein expression in comparison with normal fallopian tube non-ciliated epithelium cell line (FNE1). Nuclear localization of the PTCH1 protein in tumor tissue and cultured tumor cells suggests that this protein could play an active tumor promoter role in the nuclei of serous ovarian carcinoma cells. PTCH1 protein fragments of different molecular weights were detected in the cell lines, indicating possible proteolytic cleavage of this protein, resulting in the generation of soluble N-terminal fragments that are translocated to the nucleus. DNA methylation of the PTCH1 gene promoter was not in line with the expression level of this protein, suggesting that possibly other mechanisms, either epigenetic or posttranslational, regulate PTCH1 gene expression and protein level in serous ovarian carcinomas. Conclusions Our results indicate that PTCH1 protein could play an active tumor promoter role in the pathogenesis of serous ovarian carcinoma.

scholarly journals Molecular Characterization, Expression and Functional Analysis of Yak IFITM3 Gene

2021 ◽  
Author(s):  
Haipeng Wang ◽  
Li Wang ◽  
Juan Li ◽  
Fang Fu ◽  
Yao Zheng ◽  
...  
Keyword(s):  
Hepg2 Cells ◽  
Expression Level ◽  
Coding Region ◽  
Migration Ability ◽  
Expression Levels ◽  
Prokaryotic Expression Vector ◽  
And Migration ◽  
Lung And Kidney ◽  
P53 Genes

Abstract Background IFITM3 is interferon-induced transmembrane 3, which plays an extremely key role in anti-proliferation, anti-virus and anti-tumor diseases. To expand our understanding of the role of IFITM3 in yak, this experiment studied its function. Results Firstly, the yak ( Bos grunniens ) IFITM3 ( BgIFITM3 ) gene contained a 5’-untranslated region (UTR) (25 bp), a coding region (441 bp), and a 3’-UTR (115 bp). The expression of BgIFITM3 gene in liver was significantly higher than that in heart, spleen, lung and kidney ( P <0.01). BgIFITM3 protein was localized on the yak hepatocyte membrane, and its expression level was increased first and then stabilized from 1 day to 5 years of age. Moreover, the prokaryotic expression vector of BgIFITM3 protein was constructed and expressed successfully, with a molecular weight of 19.5 kDa. Besides, the activity of yak hepatocyte was significantly inhibited after treating with BgIFITM3 protein (10 and 20 μg/mL) ( P <0.01). The expression levels of ERBB-2, IRS-1, PI3KR-1, AKT-1 and MAPK-3 were significantly lower after treating with 20 μg/mL BgIFITM3 protein ( P <0.05). Finally, the activity of HepG2 cells was significantly inhibited after treating with BgIFITM3 protein (1, 10 and 20 μg/mL) ( P <0.05). While the cloning ability and migration ability of HepG2 cells were significantly inhibited after treating with 10 μg/mL BgIFITM3 protein ( P <0.05). The mitochondria of HepG2 cells were concentrated, cristae widened, and the double film density of mitochondria was increased after treating with 10 μg/mL BgIFITM3 protein. After 10 μg/mL BgIFITM3 protein treating, the expression levels of VDAC-2, VDAC-3 , p53 genes were significantly increased, but the expression level of GPX-4 gene was significantly decreased ( P <0.01). Conclusion Taken together, the BgIFITM3 protein could inhibit the proliferations of yak hepatocyte and HepG2 cells by regulating the PI3K/Akt pathway or ferroptosis-related genes, respectively. These results benefit for further study of the function of BgIFITM3 protein.

scholarly journals A MicroRNA-124 Polymorphism is Associated with Fracture Healing via Modulating BMP6 Expression

2017 ◽  
Vol 41 (6) ◽  
pp. 2161-2170 ◽  
Author(s):  
Lin Zou ◽  
Guichun Zhang ◽  
Lifeng Liu ◽  
Chen Chen ◽  
Xuecheng Cao ◽  
...  
Keyword(s):  
Protein Expression ◽  
Fracture Healing ◽  
Plate Osteosynthesis ◽  
Distal Tibia ◽  
Expression Level ◽  
Protein Expression Level ◽  
Regulatory Relationship ◽  
Metaphyseal Fracture ◽  
Mrna And Protein Expression ◽  
The Impact

Background: miR-124-3p has been reported to be involved in the pathogenesis of many diseases by modulating a variety of signaling pathways. In this study, we aimed to understand the impact of miR-124-3p expression level on the fracture healing in the patients of metaphyseal fracture of distal tibia, who received minimal invasive percutaneous plate osteosynthesis. Methods: We firstly collected 195 patients of metaphyseal fracture of distal tibia, and the genotype of rs531564 was determined: GG (n=124) and GC+CC (n=71). We collected information of the participants including age, gender, total in-hospital time, smoking and alcohol consumption. Subsequently, we searched the miRNA database online to identify the possible binding sequence of miR-124-3p located within the 3’-UTR of the target gene. We did correlation analysis and luciferase to understand the regulatory relationship between miR-124-3p and BMP6. Meanwhile, we also conducted real time PCR and western blotting analysis to study the mRNA and protein expression level of BMP6 in different genotype groups. We then treated the cells with scramble control, miR-124-3p mimics, BMP6 siRNA and miR-124-3p inhibitors to investigate the influence of miR-124-3p on the expression of BMP6, viability and apoptosis of cells. Results: Total in-hospital time was significantly longer in GC+CC group than GG group. MiR-124-3p was up-regulated in GG group than GC and CC groups. BMP6 was virtual target of miR-124-3p. There existed negative regulatory relationship betweenmiR-124-3p and BMP6. The mRNA and protein expression level of BMP6 decreased in GG group. MiR-124-3p decreased the expression of BMP6. MiR-124-3p negatively interfered with the viability of cells and BMP6 positively interfered with the viability of cells. MiR-124-3p reduced apoptosis and BMP6 promoted apoptosis. Conclusion: These data proved the expression of miR-124-3p was associated with the healing of metaphyseal fracture of distal tibia, and could be recognized as a biomarker to predict the healing of metaphyseal fracture of distal tibia.

scholarly journals The “Janus Face” of Platelets in Cancer

2020 ◽  
Vol 21 (3) ◽  
pp. 788 ◽  
Author(s):  
Maria Valeria Catani ◽  
Isabella Savini ◽  
Valentina Tullio ◽  
Valeria Gasperi
Keyword(s):  
Cancer Cell ◽  
Cancer Progression ◽  
Therapeutic Strategy ◽  
Cell Types ◽  
Vital Role ◽  
Bioactive Molecules ◽  
Cancer Type ◽  
Cancer Management ◽  
The Impact

Besides their vital role in hemostasis and thrombosis, platelets are also recognized to be involved in cancer, where they play an unexpected central role: They actively influence cancer cell behavior, but, on the other hand, platelet physiology and phenotype are impacted by tumor cells. The existence of this platelet-cancer loop is supported by a large number of experimental and human studies reporting an association between alterations in platelet number and functions and cancer, often in a way dependent on patient, cancer type and treatment. Herein, we shall report on an update on platelet-cancer relationships, with a particular emphasis on how platelets might exert either a protective or a deleterious action in all steps of cancer progression. To this end, we will describe the impact of (i) platelet count, (ii) bioactive molecules secreted upon platelet activation, and (iii) microvesicle-derived miRNAs on cancer behavior. Potential explanations of conflicting results are also reported: Both intrinsic (heterogeneity in platelet-derived bioactive molecules with either inhibitory or stimulatory properties; features of cancer cell types, such as aggressiveness and/or tumour stage) and extrinsic (heterogeneous characteristics of cancer patients, study design and sample preparation) factors, together with other confounding elements, contribute to “the Janus face” of platelets in cancer. Given the difficulty to establish the univocal role of platelets in a tumor, a better understanding of their exact contribution is warranted, in order to identify an efficient therapeutic strategy for cancer management, as well as for better prevention, screening and risk assessment protocols.

Synthesis of gold(iii) ← gold(i)–NHC through disproportionation: the role of gold(i)–NHC in the induction of apoptosis in HepG2 cells

2016 ◽  
Vol 40 (7) ◽  
pp. 6289-6298 ◽  
Author(s):  
Abhishek Nandy ◽  
Tapastaru Samanta ◽  
Sumana Mallick ◽  
Partha Mitra ◽  
Saikat Kumar Seth ◽  
...  
Keyword(s):  
Hepg2 Cells ◽  
Anticancer Activities ◽  
Induction Of Apoptosis

The anticancer activities of novel Au(i) and Au(iii)–NHC complexes based on 2-[(6-methylpyridin-2-yl)]imidazo[1,5-a]pyridin-4-ylium hexafluorophosphate have been investigated.

scholarly journals Thymus-Pineal Gland Axis: Revisiting Its Role in Human Life and Ageing

2020 ◽  
Vol 21 (22) ◽  
pp. 8806
Author(s):  
Rita Rezzani ◽  
Caterina Franco ◽  
Rüdiger Hardeland ◽  
Luigi Fabrizio Rodella
Keyword(s):  
Pineal Gland ◽  
Therapeutic Strategy ◽  
Human Life ◽  
Dynamic Network ◽  
Dynamical Interaction ◽  
Circadian Oscillators ◽  
The One ◽  
Definition Of ◽  
The Impact

For years the thymus gland (TG) and the pineal gland (PG) have been subject of increasingly in-depth studies, but only recently a link that can associate the activities of the two organs has been identified. Considering, on the one hand, the well-known immune activity of thymus and, on the other, the increasingly emerging immunological roles of circadian oscillators and the rhythmically secreted main pineal product, melatonin, many studies aimed to analyse the possible existence of an interaction between these two systems. Moreover, data confirmed that the immune system is functionally associated with the nervous and endocrine systems determining an integrated dynamic network. In addition, recent researches showed a similar, characteristic involution process both in TG and PG. Since the second half of the 20th century, evidence led to the definition of an effectively interacting thymus-pineal axis (TG-PG axis), but much has to be done. In this sense, the aim of this review is to summarize what is actually known about this topic, focusing on the impact of the TG-PG axis on human life and ageing. We would like to give more emphasis to the implications of this dynamical interaction in a possible therapeutic strategy for human health. Moreover, we focused on all the products of TG and PG in order to collect what is known about the role of peptides other than melatonin. The results available today are often unclear and not linear. These peptides have not been well studied and defined over the years. In this review we hope to awake the interest of the scientific community in them and in their future pharmacological applications.

PBX3 is Essential for Leukemia Stem Cell Maintenance in MLL-Rearranged Leukemia

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 2694-2694
Author(s):  
Huidong Guo ◽  
Yajing Chu ◽  
Le Wang ◽  
Hui Cheng ◽  
Weili Wang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Poor Prognosis ◽  
Hematopoietic Cells ◽  
High Expression ◽  
Expression Level ◽  
Repressive Mark ◽  
Leukemia Development ◽  
The Impact ◽  
High Expression Level

Abstract Acute myeloid leukemia (AML) with MLL translocation is recognized as a group of aggressive hematopoietic malignancies with poor prognosis. An atypical homeodomain protein family (TALE family, three-amino-acid loop extension) has been reported to be involved in leukemogenesis and leukemia progression. For example, MEIS1 and PBX3 were abnormally expressed in examined AML patient samples when compared with their normal counterparts. While MEIS1 has been studied extensively, the precise role of PBX3 in leukemia development is still largely unknown. In this study, we explored the epigenetic regulatory network of PBX3 and the specific role of PBX3 in leukemia progression. By analyzing the clinical database, we found that the high expression of PBX3 iscorrelated with poor prognosis of AML patients. In MLL-fusion induced AML mouse models that were established by retrovirus-mediated ectopic expression of MLL-AF9 or MLL-NRIP3 genes in murine BM progenitor cells (c-Kit+). PBX3 also showed high expression level in leukemia stem cells (LSC) compared with normal HSPCs. Unlike other homeobox genes such as HOXA9 and MEIS1 that were both highly expressed in LSKs and LSCs, PBX3 only showed extremely high expression in LSCs in comparison with HSPCs or other normal hematopoietic cells. To explore the role of PBX3 in leukemia, we analyzed the epigenetic modification patterns along PBX3 gene. By analyzing the public ChIP-seq database, we found that increased activation mark H3K79me2 and decreased repressive mark H3K27me3 exhibited on PBX3 in LSCs. Additionally, transcription activating modifications H3K79 hypermethylation of PBX3 was only observed in LSCs but not LSKs or GMPs, consistent with increased PBX3 expression level in LSCs compared with HSPCs. To further confirm this, we assessed H3K79me2 level of PBX3 in purified c-Kit+ cells which are enriched with stem cells from leukemic or control mice by ChIP-qPCR. Consistently, ChIP-qPCR results revealed the similar trend with dataset analysis. Moreover, we also detected the transcription repressive mark H3K9me3 on PBX3 in c-Kit+ cells and found a dramatic decrease of H3K9me3 in LSCs in comparison with HSPCs. Thus, our results suggested that high expression level of PBX3 in LSCs might be caused by elevated activation mark H3K79me2 and depressed repressive mark H3K27me3 and H3K9me3. Considering that high expression level of PBX3 in LSCs may contribute to leukemic progression, we used the CRISPR/Cas9 system to delete PBX3 to examine its potential role in MLL-AF9 mediated AML development. We found that PBX3 inactivation significantly prolonged the survival of leukemic mice (median survival was 25 days of control, 31 days of SgPBX3-a, 30 days of SgPBX3-b, 10 mice per group). Morphological analysis revealed more leukemic blast cells with segment nuclei exhibited in PBX3 inactivation group, suggesting that PBX3 deficiency is associated with increased leukemic cell differentiation. Since LSC was considered to be sufficient to drive leukemogenesis and maintain leukemia progression, we investigated the impact of PBX3 inactivation on LSC capability. In vitro colony forming assay showed that PBX3 inactivation group formed significantly fewer colonies of total numbers and type A clones. Flow cytometric analysis showed decreased LSCs frequency in murine leukemia cells of PBX3 inactivation group. Moreover, limiting dilution transplantation assays revealed 4-7 fold decreased of functional LSCs after PBX3 inactivation with the 1/102 of LSCs in control leukemic bone marrow in comparison with 1/442 in SgPBX3-a and 1/741 in SgPBX3-b respectively. Further analysis suggested that depletion of functional LSCs in PBX3 inactivation group mainly through accelerating apoptosis of LSCs, evidenced by both increased Annexin V positive cell frequency of whole BM blasts and LSCs in PBX3 inactivation group. In conclusion, we found PBX3 is epigenetically dis-regulated in LSCs of MLL-r AMLs and is essential for leukemia development. Inactivation of PBX3 could suppress leukemia progression by reducing LSCs frequency and impairing its function. More importantly, the differential expression of PBX3 in normal and malignant hematopoietic cells provides itself potentially as a prognostic marker and therapeutic target for MLL-rearranged leukemia. HD.G and YJ.C contributed equally to this work Disclosures No relevant conflicts of interest to declare.

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