scholarly journals Increased IL-1a expression is correlated with bladder cancer malignant progression

2020 ◽  
Author(s):  
Shi-Jie Yao ◽  
Hong-Shun Ma ◽  
Guang-Ming Liu ◽  
Yue Gao ◽  
Wei Wang
Keyword(s):  
Bladder Cancer ◽  
Protein Expression ◽  
Western Blot ◽  
Quantitative Polymerase Chain Reaction ◽  
Malignant Progression ◽  
Nuclear Antigen ◽  
Ki 67 ◽  
Gene And Protein Expression

IntroductionTo explore the function of interleukin 1α (IL-1α) in bladder cancer (BCa).Material and methodsImmunohistochemistry (IHC) was used to test the protein expression of IL-1α in BCa tissues. The relationship between IL-1α and clinical characteristics was analyzed by the Kaplan-Meier curve method. The gene and protein expression was tested by reverse transcription quantitative polymerase chain reaction (RT-q-PCR) and western blot, respectively. Colony formation and MTT assays were used to detect the potential of proliferation in vitro, and scratch and transwell chamber assays were used to detect the potential of invasion in vitro. Markers of proliferation such as Ki-67 and proliferating cell nuclear antigen (PCNA) and markers of invasion such as MMP-2 and MMP-9 were detected by western blot. Xenograft study was used for the in vivo experiment.ResultsWe found that IL-1α was highly expressed in BCa patients while highly expressed IL-1α was significantly related to short overall survival and progression-free survival in BCa as well. Moreover, knockdown of IL-1α might inhibit the ability of cancer cells to proliferate and invade or migrate both in vitro and in vivo.ConclusionsOur findings suggested that IL-1α might be a therapy target for BCa malignant progression.

Effect of Auraptene on angiogenesis in Xenograft model of breast cancer

2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Mohammad Reza Shiran ◽  
Elham Mahmoudian ◽  
Abolghasem Ajami ◽  
Seyed Mostafa Hosseini ◽  
Ayjamal Khojasteh ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Gene Expression ◽  
Animal Model ◽  
Protein Expression ◽  
Western Blot ◽  
Xenograft Model ◽  
Dependent Manner ◽  
Concentration Dependent Manner

Abstract Objectives Angiogenesis is the most important challenge in breast cancer treatment. Recently, scientists become interesting in rare natural products and intensive researches was performed to identify their pharmacological profile. Auraptene shows helpful effects such as cancer chemo-preventive, anti-inflammatory, anti-oxidant, immuno-modulatory. In this regard, we investigated the anti-angiogenesis effect of Auraptene in in-vitro and in-vivo model of breast cancer. Methods In this study, 4T, MDA-MB-231 and HUVEC cell lines were used. The proliferation study was done by MTT assay. For tube formation assay, 250 matrigel, 1 × 104 HUVEC treated with Auraptene, 20 ng/mL EGF, 20 ng/mL bFGF and 20 ng/mL VEGF were used. Gene expression of important gene related to angiogenesis in animal model of breast cancer was investigated by Real-time PCR. Protein expression of VCAM-1 and TNFR-1 gene related to angiogenesis in animal model of breast cancer was investigated by western-blot. Results Auraptene treatment led to reduction in cell viability of MDA-MB-231 in a concentration-dependent manner. Also, we observed change in the number of tubes or branches formed by cells incubated with 40 and 80 μM Auraptene. Auraptene effect the gene expression of important gene related to angiogenesis (VEGF, VEGFR2, COX2, IFNɣ). Moreover, the western blot data exhibited that Auraptene effect the protein expression of VCAM-1 and TNFR-1. Conclusions Overall, this study shows that Auraptene significantly suppressed angiogenesis via down-regulation of VEGF, VEGFR2, VCAM-1, TNFR-1, COX-2 and up-regulation of IFNγ.

scholarly journals In Vitro and In Vivo Effects of Fermented Oyster-Derived Lactate on Exercise Endurance Indicators in Mice

2020 ◽  
Vol 17 (23) ◽  
pp. 8811
Author(s):  
Storm N. S. Reid ◽  
Joung-Hyun Park ◽  
Yunsook Kim ◽  
Yi Sub Kwak ◽  
Byeong Hwan Jeon
Keyword(s):  
Lactate Dehydrogenase ◽  
Protein Expression ◽  
Western Blot ◽  
Blot Analysis ◽  
Western Blot Analysis ◽  
Biochemical Analysis ◽  
Positive Control ◽  
Exercise Endurance

Exogenous lactate administration has more recently been investigated for its various prophylactic effects. Lactate derived from potential functional foods, such as fermented oyster extract (FO), may emerge as a practical and effective method of consuming exogenous lactate. The current study endeavored to ascertain whether the lactate derived from FO may act on muscle cell biology, and to what extent this may translate into physical fitness improvements. We examined the effects of FO in vitro and in vivo, on mouse C2C12 cells and exercise performance indicators in mice, respectively. In vitro, biochemical analysis was carried out to determine the effects of FO on lactate content and muscle cell energy metabolism, including adenosine triphosphate (ATP) activity. Western blot analysis was also utilized to measure the protein expression of total adenosine monophosphate-activated protein kinase (AMPK), p-AMPK (Thr172), lactate dehydrogenase (LDH), succinate dehydrogenase (SDHA) and peroxisome proliferator-activated receptor gamma coactivator-1α (PGC-1α) in response to FO administration. Three experimental groups were formed: a positive control (PC) treated with 1% horse serum, FO10 treated with 10 μg/mL and FO50 treated with 50 μg/mL. In vivo, the effects of FO supplementation on exercise endurance were measured using the Rota-rod test, and Western blot analysis measured myosin heavy-chain 2 (MYH2) to assess skeletal muscle growth, alongside p-AMPK, total-AMPK, PGC-1α, cytochrome C and UCP3 protein expression. Biochemical analysis was also performed on muscle tissue to measure the changes in concentration of liver lactate, lactate dehydrogenase (LDH), glycogen and citrate. Five groups (n = 10/per group) consisted of a control group (CON), exercise group (Ex), positive control treated with Ex and 500 mg/kg Taurine (Ex-Tau), Ex and 100 mg/kg FO supplementation (Ex-FO100) and Ex and 200 mg/kg FO supplementation (Ex-FO200) orally administered over the 4-week experimental period.FO50 significantly increased PGC-1α expression (p < 0.001), whereas both FO10 and FO50 increased the expression of p-AMPK (p < 0.001), in C2C12 muscle cells, showing increased signaling important for mitochondrial metabolism and biogenesis. Muscle lactate levels were also significantly increased following FO10 (p < 0.05) and FO50 (p < 0.001). In vivo, muscle protein expression of p-AMPK (p < 0.05) and PGC-1α were increased, corroborating our in vitro results. Cytochrome C also significantly increased following FO200 intake. These results suggest that the effects of FO supplementation may manifest in a dose-response manner. FO administration, in vitro, and supplementation, in vivo, both demonstrate a potential for improvements in mitochondrial metabolism and biogenesis, and even for potentiating the adaptive effects of endurance exercise. Mechanistically, lactate may be an important molecule in explaining the aforementioned positive effects of FO.

scholarly journals Alcohol induces TGFβ1 via downregulation of miR-1946a in murine lung fibroblast

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Xian Fan ◽  
Stephen T. Mills ◽  
Mevelyn J. Kaalla ◽  
Viranuj Sueblinvong
Keyword(s):  
Protein Expression ◽  
Transforming Growth Factor Beta ◽  
Transforming Growth Factor ◽  
Smooth Muscle Actin ◽  
Direct Interaction ◽  
Lung Fibroblasts ◽  
Murine Lung ◽  
Gene And Protein Expression

Abstract Exaggerated transforming growth factor-beta 1 (TGFβ1) expression worsens fibroproliferation following bleomycin-induced lung injury in alcohol-fed mice. MicroRNA (miR)-1946a is predicted to bind to the TGFβ1 3′ untranslated region (UTR), thereby inhibiting its transcription. We hypothesize that alcohol suppresses miR-1946a and induces TGFβ1. Primary murine lung fibroblasts (PLFs) were cultured ± alcohol, miR-1946a mimic or inhibitor, and TGFβ1 signaling inhibitors. miR-1946a was analyzed after alcohol treatment in vitro and in vivo. TGFβ1 expression and TGFβ1 3′UTR-luciferase activity was quantified. We showed that alcohol suppressed miR-1946a in the alcohol-fed mouse lungs and PLFs. MiR-1946a inhibitor increased TGFβ1 expression in the fibroblast. MiR-1946a mimic treatment suppressed TGFβ1 gene expression and TGFβ1 3′UTR activity. Overexpression of miR1946a inhibited alcohol-induced TGFβ1 gene and protein expression as well as alcohol-induced TGFβ1 and α-smooth muscle actin (SMA) protein expression in PLFs. In conclusion, miR-1946a modulates TGFβ1 expression through direct interaction with TGFβ1 3′UTR. These findings identify a novel mechanism by which alcohol induces TGFβ1 in the lung.

3 SIRT1—A POSSIBLE MARKER FOR REPRODUCTIVE AGING OF IN VIVO-DERIVED BOVINE OOCYTES?

2017 ◽  
Vol 29 (1) ◽  
pp. 109 ◽  
Author(s):  
P. Kordowitzki ◽  
S. Klein ◽  
K.-G. Hadeler ◽  
P. Aldag ◽  
M. Nowak-Imialek ◽  
...  
Keyword(s):  
Protein Expression ◽  
Expression Profiles ◽  
Reproductive Aging ◽  
Standard Protocol ◽  
Blastocyst Formation ◽  
Fluorescent Intensity ◽  
Gene And Protein Expression ◽  
Maternal Aging

Maternal aging-associated reduction of oocyte viability is a common feature in mammals. Effective measures to counteract this process have not yet been developed. Cows are commonly used as a model of early human development, including maternal aging, because both species share a very high degree of similarity, including follicle selection, cleavage and blastocyst formation and a long reproductive lifespan. SIRT1, a member of the Sirtuin family, deacetylates transcriptional regulators localised in the nucleus and cytoplasm by a NAD+-dependent mechanism. Resveratrol (3,4′,5-trihydroxystilbene) is an antioxidant identified in various plant species and red wine which enhances SIRT1 activity. Based on these observations, the goal of the present study was to examine, if SIRT1 gene and protein expression is either affected by maternal age and/or can be modulated by resveratrol. Cumulus-oocyte-complexes of prepubertal (5–6 months old) and adult/aged (2 to 8 lactation) cows were collected by ovum pick-up twice a week. Medium for in vitro maturation (TCM 199) and in vitro fertilization (FertTalp) was supplemented with 20 µL of Resveratrol® (Sigma-Aldrich, Buchs, Switzerland) to get a final concentration of 2 µM Resveratrol respectively. Standard (TCM 199 and FertTalp) media without Resveratol were used as control. Cleavage rates and blastocyst formation were evaluated. Comprehensive gene expression assays of germinal vesicle and metaphase II (MII)-stage oocytes and blastocyst were conducted using next-generation sequencing technology. Finally, SIRT1 protein expression in oocytes and blastocysts were analysed by fluorescence immunostaining under a confocal microscope (LSM510, Zeiss, Germany) and relative fluorescent intensity was calculated. The cleavage rates of adult and prepubertal donors did not differ significantly among the treatments (standard protocol: 56.5 ± 5.4% for adult and 53.0 ± 4.7% for prepubertal donors, Resveratrol supplemented protocol: 62.1 ± 4.3% for cows and 63.6 ± 3.9% for calves). The blastocyst rates were slightly enhanced in the Resveratrol supplemented groups (cows: 34.2 ± 3.8% and calves: 33.1 ± 4.2%) compared to those of standard protocol (cows: 27.5 ± 4.8% and calves: 26.4 ± 3.3%). Relative mRNA abundance levels of SIRT1 were lower in oocytes and blastocysts derived from cows than in those derived from their younger counterparts (2.8-fold change; P = 0.05), but did not differ significantly among treatment groups. Protein expression profiles revealed that bovine SIRT1 was localised in the nucleus. The relative fluorescence levels of SIRT1 were significantly lower (221 ± 34 FIU) in control groups compared to the resveratrol treated groups (865 ± 45 FIU, respectively; P = 0.05). Additionally, SIRT1 protein levels were significantly higher in MII-oocytes (1255 ± 56 FIU) and blastocysts (984 ± 26 FIU) derived from calves compared with their older counterparts (442 ± 37 FIU and 310 ± 23 FIU, respectively, P = 0.05). In conclusion, these results indicate that resveratrol affects SIRT1 protein expression in oocytes and blastocysts of donors in different age. Thus, we hypothesise that SIRT1 is a reliable marker for reproductive aging, which could also be useful for better understanding of human infertility caused by aging.

Deregulated Expression of a Novel Oncogene, Ahi-1, in Murine Hematopoietic Cells Confers a Proliferative Advantage In Vitro and Leukemogenic Activity In Vivo and Enhances the Effects of BCR-ABL.

Blood ◽  
2006 ◽  
Vol 108 (11) ◽  
pp. 388-388
Author(s):  
Leon Zhou ◽  
Ashley Ringrose ◽  
Ann E.-J. Lin ◽  
Xiaoyan Jiang
Keyword(s):  
Growth Factor ◽  
Protein Expression ◽  
Western Blot ◽  
Proliferative Activity ◽  
Integration Site ◽  
Hematopoietic Cells ◽  
Leukemic Cells ◽  
Biological Behavior

Abstract Ahi-1 (Abelson helper integration site-1) is a novel gene that we recently identified based on its common activation in v-abl or myc-induced murine leukemias and lymphomas. It encodes a unique protein with known signaling features, including SH3 and WD40-repeat domains, but its function is largely unknown. We have recently demonstrated that Ahi-1/AHI-1 transcript levels are normally down-regulated during both early murine and human hematopoietic cell differentiation and are highly increased in human leukemic cells, particularly in highly enriched populations of primitive BCR-ABL+ leukemic stem cells (lin−CD34+CD38−) in patients with chronic myeloid leukemia (CML). To investigate the potential cooperative activity of Ahi-1 in BCR-ABL-mediated signal transduction and leukemogenesis, we transduced IL-3-dependent BaF3 cells with MSCV-Ahi-1-IRES-YFP and/or MSCV-BCR-ABL-IRES-GFP retroviruses and compared the biological behavior of these cells in vitro and in vivo. All Ahi-1-transduced clonal cell lines showed increased proliferative activity and reduced apoptosis in the absence of IL-3, compared to parental BaF3 cells or control GFP-transduced cells. Interestingly, overexpression of both Ahi-1 and BCR-ABL caused more enhanced perturbations when compared to cells transduced with either Ahi-1 or BCR-ABL alone. Strikingly, intravenous injection of NOD/SCID-β2microglobulin−/−mice with BaF3 cells induced to overexpress Ahi-1 alone induces a lethal leukemia within 70 days. These leukemogenic activities were further increased by introduction of co-transduced Ahi-1 and BCR-ABL cells, producing a shorter latency of 30 days. A disease latency of 40 days was revealed by introduction of BCR-ABL-transduced cells alone. Western blot analysis showed that both protein expression and the tyrosine kinase activity of p210BCR-ABL were highly increased in cells co-transduced with Ahi-1 and BCR-ABL compared to BaF3 cells transduced with BCR-ABL alone. Similarly, we also observed higher levels of Ahi-1 protein expression in the same dually transduced cells (Ahi-1+BCR-ABL+) than in those transduced with Ahi-1 alone. We further demonstrated a similarly perturbed proliferative activity, growth factor independence and colony-forming cell (CFC) output in long-term culture initiated cell (LTC-IC) assays of 5-fluorouracil (5-FU)-treated primitive murine BM cells (Lin−Sca1+ cells) transduced with Ahi-1 and BCR-ABL, either alone or in combination. To investigate directly the cooperating oncogenic role of AHI-1 in BCR-ABL-mediated malignant transformation of CML cells, knockdown of AHI-1 expression in K562 cells, a cell line that was derived from a patient with CML and that is characterized by highly increased expression of AHI-1, was performed using retroviral-mediated RNA interference. Retroviral-mediated suppression specifically inhibited endogenous AHI-1 expression in transduced cells by 70% as evaluated by Q-RT-PCR and Western blot analyses. It further caused a significant reduction in their growth factor independence in semi-solid cultures (up to 5-fold) and in single cell cultures (2-fold) by comparison to cells transduced with a control vector. Taken together, these findings provide strong evidence of the transforming potential of Ahi-1/AHI-1 in primitive hematopoietic cells. This effect is additive with those of BCR-ABL, suggesting that AHI-1 and BCR-ABL can play a cooperate role in the development of BCR-ABL-associated diseases like CML.

scholarly journals Region between the Canine Distemper Virus M and F Genes Modulates Virulence by Controlling Fusion Protein Expression

2008 ◽  
Vol 82 (21) ◽  
pp. 10510-10518 ◽  
Author(s):  
Danielle E. Anderson ◽  
Veronika von Messling
Keyword(s):  
Protein Expression ◽  
Measles Virus ◽  
Canine Distemper Virus ◽  
Transcriptional Control ◽  
Canine Distemper ◽  
F Protein ◽  
F Gene ◽  
Gene And Protein Expression

ABSTRACT Morbilliviruses, including measles and canine distemper virus (CDV), are nonsegmented, negative-stranded RNA viruses that cause severe diseases in humans and animals. The transcriptional units in their genomes are separated by untranslated regions (UTRs), which contain essential transcription and translation signals. Due to its increased length, the region between the matrix (M) protein and fusion (F) protein open reading frames is of particular interest. In measles virus, the entire F 5′ region is untranslated, while several start codons are found in most other morbilliviruses, resulting in a long F protein signal peptide (Fsp). To characterize the role of this region in morbillivirus pathogenesis, we constructed recombinant CDVs, in which either the M-F UTR was replaced with that between the nucleocapsid (N) and phosphoprotein (P) genes, or 106 Fsp residues were deleted. The Fsp deletion alone had no effect in vitro and in vivo. In contrast, substitution of the UTR was associated with a slight increase in F gene and protein expression. Animals infected with this virus either recovered completely or experienced prolonged disease and death due to neuroinvasion. The combination of both changes resulted in a virus with strongly increased F gene and protein expression and complete attenuation. Taken together, our results provide evidence that the region between the morbillivirus M and F genes modulates virulence through transcriptional control of the F gene expression.

scholarly journals Effect of ovarian steroids on vascular endothelial growth factor a expression in bovine uterine endothelial cells during adenomyosis

2019 ◽  
Vol 15 (1) ◽  
Author(s):  
Martyna Lupicka ◽  
Anna Zadroga ◽  
Agata Szczepańska ◽  
Anna Justyna Korzekwa
Keyword(s):  
Vascular Endothelial Growth Factor ◽  
Endothelial Cells ◽  
Growth Factor ◽  
Protein Expression ◽  
Vascular Endothelial Cells ◽  
Quantitative Polymerase Chain Reaction ◽  
Vascular Endothelial ◽  
Gene And Protein Expression ◽  
Endothelial Growth Factor

Abstract Background Adenomyosis is a uterine dysfunction defined as the presence of endometrial glands within the myometrium. There is evidence that proangiogenic factors may play a role during the development of adenomyosis; however, exact mechanism remains unknown. The aim of the study was to determine the action of vascular endothelial growth factor A (VEGFA) in uterine tissue and uterine vascular endothelial cells during adenomyosis. Results Uterine tissues were collected and examined for the presence and extent of adenomyosis. Gene and protein expression of VEGFA and its two receptors (VEGFR1 and VEGFR2) was evaluated with quantitative polymerase chain reaction and Western blotting, respectively, in endometrium and myometrium during adenomyosis. Immunolocalization of VEGFA and its receptors within uterine tissues during adenomyosis was also determined. In an in vitro experiment, endothelial cells from non-adenomyotic bovine uteri were treated with media conditioned by non-adenomyotic or adenomyotic uterine slices treated with 17-beta-oestradiol (E2) or progesterone (P4). Both gene and protein expression of VEGFR2 were elevated in endometrium in stages 3–4 of adenomyosis. Protein expression of VEGFA and VEGFR2 as well as VEGFA secretion were increased in endothelial cells treated with media conditioned by adenomyotic uterine slices after E2 treatment. Conclusions Results suggest that VEGFA signalling is an important component, next to E2, that enhances VEGFA action and participates in adenomyosis development in cows.

scholarly journals YTHDF2 facilitates UBXN1 mRNA decay by recognizing METTL3-mediated m6A modification to activate NF-κB and promote the malignant progression of glioma

2021 ◽  
Vol 14 (1) ◽  
Author(s):  
Rui-Chao Chai ◽  
Yu-Zhou Chang ◽  
Xin Chang ◽  
Bo Pang ◽  
Song Yuan An ◽  
...  
Keyword(s):  
Rna Binding ◽  
Molecular Subtype ◽  
Quantitative Polymerase Chain Reaction ◽  
Rna Stability ◽  
Malignant Progression ◽  
Mouse Tumor ◽  
Mrna Levels ◽  
In Vivo Models

Abstract Background The prognosis for diffuse gliomas is very poor and the mechanism underlying their malignant progression remains unclear. Here, we aimed to elucidate the role and mechanism of the RNA N6,2′-O-dimethyladenosine (m6A) reader, YTH N6-methyladenosine RNA binding protein 2 (YTHDF2), in regulating the malignant progression of gliomas. Methods YTHDF2 mRNA levels and functions were assessed using several independent datasets. Western blotting, quantitative polymerase chain reaction, and immunohistochemistry were used to evaluate the expression levels of YTHDF2 and other molecules in human and mouse tumor tissues and cells. Knockdown and overexpression were used to evaluate the effects of YTHDF2, methyltransferase-like 3 (METTL3), and UBX domain protein 1 (UBXN1) on glioma malignancy in cell and orthotopic xenograft models. RNA immunoprecipitation (RIP), methylated RIP, and RNA stability experiments were performed to study the mechanisms underlying the oncogenic role of YTHDF2. Results YTHDF2 expression was positively associated with a higher malignant grade and molecular subtype of glioma and poorer prognosis. YTHDF2 promoted the malignant progression of gliomas in both in vitro and in vivo models. Mechanistically, YTHDF2 accelerated UBXN1 mRNA degradation via METTL3-mediated m6A, which, in turn, promoted NF-κB activation. We further revealed that UBXN1 overexpression attenuated the oncogenic effect of YTHDF2 overexpression and was associated with better survival in patients with elevated YTHDF2 expression. Conclusions Our findings confirmed that YTHDF2 promotes the malignant progression of gliomas and revealed important insight into the upstream regulatory mechanism of NF-κB activation via UBXN1 with a primary focus on m6A modification.

scholarly journals miR-3687 Overexpression Promotes Bladder Cancer Cell Growth Through Inhibiting the Negative Effect of FOXP1 on Cyclin E2 Transcription

2018 ◽  
Author(s):  
Qipeng Xie ◽  
Caiyi Chen ◽  
Haiying Li ◽  
Huiying Yan ◽  
Jiheng Xu ◽  
...  
Keyword(s):  
Cell Cycle ◽  
Bladder Cancer ◽  
Protein Expression ◽  
Cancer Progression ◽  
Cancer Cell Growth ◽  
Cycle Arrest ◽  
Cyclin E2 ◽  
Negative Effect

Cyclin E2, a member of the cyclin family, is a key cell cycle-related protein. This protein plays essential roles in cancer progression, and as such, an inhibitor of cyclin E2 has been approved to treat several types of cancers. Even so, mechanisms underlying how to regulate cyclin E2 expression in cancer remain largely unknown. The current study found that miR-3687 is up-regulated in clinical bladder cancer (BC) tumor tissues, TCGA database and human BC cell lines. Inhibition of miR-3687 expression significantly reduces human BC cell proliferation in vitro and tumor growth in vivo, which are concurrently with the induction of G0/G1 cell cycle arrest and downregulation of cyclin E2 protein expression. Interestingly, overexpression of cyclin E2 reversed the inhibition of BC proliferation induced by miR-3687. Mechanistic studies suggest that miR-3687 could bind to the 3'-UTR of foxp1 mRNA, downregulates FOXP1 protein expression, and in turn promotes the transcription of cyclin E2, thereby promoting the growth of BC cells. Collectively, the current study not only establishes a novel regulatory axis of miR-3687/FOXP1 in regard to regulation of cyclin E2 expression in BC cells, but also provides strong suggestive evidence that miR-3687 and FOXP1 may be potentially promising targets in therapeutic strategies of human BC.

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