Expression of Notch–Hif-1α signaling pathway in liver regeneration of rats

Objective To investigate whether the Notch–Hif-1α signaling pathway is involved in liver regeneration. Methods Rats were divided into two groups and treated with daily intraperitoneal injections of saline (control) or the gamma-secretase inhibitor, Fli-06, for 2 days. Two-thirds of the rat livers were resected and rats were later euthanized at specific time points post-resection to analyze the remnant livers. Each group's liver/body weight ratio was calculated, and immunostaining and western blotting were used to determine the cell proliferation marker, PCNA and Ki-67 expression. Real-time PCR and western blotting were used to compare the mRNA expression of Notch homolog-1 ( Notch1), hairy and enhancer of split-1 ( Hes1), and vascular endothelial growth factor ( Vegf), and the protein expression of NICD and HIF-1α, respectively. Results The liver/body weight ratios and number of Ki-67- and PCNA-positive cells were significantly lower in the experimental group than the control group, indicating lower levels of liver regeneration following the disruption of Notch signaling by Fli-06. The Hes1 and Vegf mRNA levels and NICD and HIF-1α protein expression levels were all down-regulated by Fli-06 treatment. Conclusion Notch–Hif-α signaling pathway activation plays an important role in liver regeneration, where it may contribute toward liver cell proliferation.

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MicroRNA-1297 suppressed the Akt/GSK3β signaling pathway and stimulated neural apoptosis in an in vivo sevoflurane exposure model

Journal of International Medical Research ◽

10.1177/0300060520982104 ◽

2021 ◽

Vol 49 (4) ◽

pp. 030006052098210

Author(s):

Quan Wang ◽

Jingcong Luo ◽

Ruiqiang Sun ◽

Jia Liu

Keyword(s):

Protein Expression ◽

Signaling Pathway ◽

Neuronal Apoptosis ◽

In Vitro Model ◽

Nervous System Development ◽

Exposure Model ◽

Control Group ◽

Pten Protein ◽

Vitro Model

Objective Common inhalation anesthetics used for clinical anesthesia (such as sevoflurane) may induce nerve cell apoptosis during central nervous system development. Furthermore, anesthetics can produce cognitive impairments, such as learning and memory impairments, that continue into adulthood. However, the precise mechanism remains largely undefined. We aimed to determine the function of microRNA-1297 (miR-1297) in sevoflurane-induced neurotoxicity. Methods Reverse transcription-polymerase chain reaction assays were used to analyze miR-1297 expression in sevoflurane-exposed mice. MTT and lactate dehydrogenase (LDH) assays were used to measure cell growth, and neuronal apoptosis was analyzed using flow cytometry. Western blot analyses were used to measure PTEN, PI3K, Akt, and GSK3β protein expression. Results In sevoflurane-exposed mice, miR-1297 expression was up-regulated compared with the control group. MiR-1297 up-regulation led to neuronal apoptosis, inhibition of cell proliferation, and increased LDH activity in the in vitro model of sevoflurane exposure. MiR-1297 up-regulation also suppressed the Akt/GSK3β signaling pathway and induced PTEN protein expression in the in vitro model. PTEN inhibition (VO-Ohpic trihydrate) reduced PTEN protein expression and decreased the effects of miR-1297 down-regulation on neuronal apoptosis in the in vitro model. Conclusion Collectively, the results indicated that miR-1297 stimulates sevoflurane-induced neurotoxicity via the Akt/GSK3β signaling pathway by regulating PTEN expression.

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miR-636 inhibits EMT, cell proliferation and cell cycle of ovarian cancer by directly targeting transcription factor Gli2 involved in Hedgehog pathway

Cancer Cell International ◽

10.1186/s12935-020-01725-7 ◽

2021 ◽

Vol 21 (1) ◽

Author(s):

Jiong Ma ◽

Chunxia Zhou ◽

Xuejun Chen

Keyword(s):

Cell Cycle ◽

Cell Proliferation ◽

Protein Expression ◽

Signaling Pathway ◽

Luciferase Reporter ◽

Cell Proliferation And Migration ◽

Hh Signaling ◽

And Migration ◽

Proliferation And Migration

Abstract Background Hedgehog (Hh) signaling pathway, which is essential for cell proliferation and differentiation, is noted to be aberrantly activated in tumor from increasing studies in recent years. MicroRNAs (miRNAs) as an important non-coding RNA in cells have been proven to possess a regulatory role specific to the Hh signaling pathway. Here, in vitro and in vivo cellular/molecular experiments were adopted to clarify the regulatory mechanism linking miR-636 to the Hh signaling pathway in ovarian cancer (OVC). Methods Protein–protein interaction analysis was performed to identify the hub gene in the Hh pathway. TargetScan database was used to predict the potential upstream regulators for Gli2. qRT-PCR was performed to test the expression of miR-636, while Western blot was conducted to detect the expression of proteins related to the Hh pathway and epithelial-mesenchymal transition (EMT). For cell functional experiments, HO-8910PM OVC cell line was used. MTT assay and wound healing assay were used to measure the effect of miR-636 on cell proliferation and migration. Flow cytometry was carried out to examine the effect of miR-636 on cell cycle, and Western blot was used to identify the change in expression of Hh and EMT-related proteins. Dual-luciferase reporter gene assay was implemented to detect the targeting relationship between miR-636 and Gli2. Xenotransplantation models were established for in vivo examination. Results Gli2 was identified as the hub gene of the Hh pathway and it was validated to be regulated by miR-636 based on the data from TargetScan and GEO databases. In vitro experiments discovered that miR-636 was significantly lowly expressed in OVC cell lines, and overexpressing miR-636 significantly inhibited HO-8910PM cell proliferation, migration and induced cell cycle arrest in G0/G1 phase, while the inhibition of miR-636 caused opposite results. Dual-luciferase reporter gene assay revealed that Gli2 was the target gene of miR-636 in OVC. Besides, overexpressed miR-636 decreased protein expression of Gli2, and affected the expression of proteins related to the Hh signaling pathway and EMT. Rescue experiments verified that overexpression of Gli2 reversed the inhibitory effect of miR-636 on HO-8910PM cell proliferation and migration, and attenuated the blocking effect of miR-636 on cell cycle. The xenotransplantation experiment suggested that miR-636 inhibited cell growth of OVC by decreasing Gli2 expression. Besides, overexpressing Gli2 potentiated the EMT process of OVC cells via decreasing E-cadherin protein expression and increasing Vimentin protein expression, and it reversed the inhibitory effect of miR-636 on OVC cell proliferation in vivo. Conclusion miR-636 mediates the activation of the Hh pathway via binding to Gli2, thus inhibiting EMT, suppressing cell proliferation and migration of OVC. Trial registration: The experimental protocol was established, according to the ethical guidelines of the Helsinki Declaration and was approved by the Human Ethics Committee of The Second Affiliated hospital of Zhejiang University School of Medicine (IR2019001235). Written informed consent was obtained from individual or guardian participants.

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Phenobarbital inhibits calpain activity and expression in mouse hepatoma cells

Biological Chemistry ◽

10.1515/hsz-2015-0223 ◽

2016 ◽

Vol 397 (1) ◽

pp. 91-96 ◽

Cited By ~ 5

Author(s):

Nicola Groll ◽

Ferdinand Kollotzek ◽

Jens Goepfert ◽

Thomas O. Joos ◽

Michael Schwarz ◽

...

Keyword(s):

Cell Proliferation ◽

Transcriptional Regulation ◽

Antiepileptic Drug ◽

Signaling Pathway ◽

Constitutive Androstane Receptor ◽

Hepatoma Cells ◽

Liver Cells ◽

Mrna Levels ◽

Calpain Activity ◽

Mouse Hepatoma

Abstract The antiepileptic drug phenobarbital (PB) exerts hepatic effects related to cell proliferation and tumorigenesis which are closely linked to the Wnt/β-catenin signaling pathway. This pathway is, amongst others, regulated by calpain proteases. We now identified PB as an inhibitor of Wnt/β-catenin signaling in mouse hepatoma cells. Further analyses revealed that PB inhibits calpain activity, an effect which is at least in parts mediated by a transcriptional regulation of calpain mRNA levels and which is furthermore independent of the constitutive androstane receptor, the known mediator of most effects of PB in liver cells.

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Evaluation of MACF1-regulatory non-coding RNA as a potential therapeutic target in Colorectal Cancer

QJM ◽

10.1093/qjmed/hcab088.011 ◽

2021 ◽

Vol 114 (Supplement_1) ◽

Author(s):

Rowaida Mohammed Reda M. M Aboushahba ◽

Fayda Ibrahim Abdel Motaleb ◽

Ahmed Abdel Aziz Abou-Zeid ◽

Enas Samir Nabil ◽

Dalia Abdel-Wahab Mohamed ◽

...

Keyword(s):

Colorectal Cancer ◽

Cell Proliferation ◽

Wnt Signaling ◽

Cell Line ◽

Signaling Pathway ◽

Therapeutic Target ◽

Molecular Mechanisms ◽

Wnt Signaling Pathway ◽

Control Group ◽

Potential Therapeutic Target

ABSTRACT Colorectal cancer (CRC) is one of the leading causes of cancer-related deaths world-wide. There is an increasing need for the identification of novel biomarkers/targets for early diagnosis and for the development of novel chemopreventive and therapeutic agents for CRC. Recently, MACF1 gene has emerged as a potential therapeutic target in cancer as it involved in processes critical for tumor cell proliferation, invasion and metastasis. It is suggested that MACF1 may function in cancers through Wnt signaling. MiR-34a is a well-known tumor suppressor miRNA.miR-34a targets MACF1 gene as a part of the wnt signaling pathway. In this study, 40 colonic tissues were collected from CRC patients (20) and control subjects (20). miR-34a-5p was assessed by real time PCR in all study groups. The results showed highly significant decrease (P < 0.01) in miR-34a relative expression in the CRC group (median RQ 0.13) when compared to the benign group (median RQ 5.3) and the healthy control group (median RQ 19.63). miR-34a mimic and inhibitor were transfected in CaCo-2 cell line and proliferation was assessed. The transfection of the cell line with miR-34a mimic decreased cell proliferation. Our study suggests that miR-34a-5p targets MACF1 gene as a part of the wnt signaling pathway leading to the involvement in the molecular mechanisms of CRC development and progression.

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Influence of Portosystemic Shunt on Liver Regeneration after Hepatic Resection in Pigs

HPB Surgery ◽

10.1155/2009/835965 ◽

2009 ◽

Vol 2009 ◽

pp. 1-8 ◽

Cited By ~ 1

Author(s):

R. Ladurner ◽

M. Schenk ◽

R. Margreiter ◽

F. Offner ◽

A. Königsrainer

Keyword(s):

Liver Regeneration ◽

Portosystemic Shunt ◽

Control Group ◽

Minimal Amount ◽

Remnant Liver ◽

Ki 67 ◽

Tissue Sections ◽

Hepatocyte Regeneration ◽

Shunt Group ◽

Left Hemihepatectomy

Objective. The minimal amount of liver mass necessary for regeneration is still a matter of debate. The aim of the study was to analyze liver regeneration factors after extended resection with or without portosystemic shunt. Methods. An extended left hemihepatectomy was performed in 25 domestic pigs, in 15 cases after a portosystemic H-shunt. The expression of Ki-67, VEGF, TGF-, FGF, and CK-7 was analyzed in paraffin-embedded tissue sections. Results. The volume of the remnant liver increased about 2.5-fold at the end of the first week after resection. With 19 cells/10 Glisson fields versus 4/10, Ki-67-expression was significantly higher in the H-shunt group. VEGF- and CK-7-expressions were significantly higher in the control group. No significant change was found in FGF-expression. The expression of TGF- was higher, but not significantly, in the control group. Conclusions. The expression of Ki-67, and therefore hepatocyte regeneration, was increased in the shunt group. The expression of CK-7 on biliary epithelium and the expression of VEGF, however, were stronger in the control group.

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miR-195 Regulates Proliferation and Apoptosis through Inhibiting the mTOR/p70s6k Signaling Pathway by Targeting HMGA2 in Esophageal Carcinoma Cells

Disease Markers ◽

10.1155/2017/8317913 ◽

2017 ◽

Vol 2017 ◽

pp. 1-9 ◽

Cited By ~ 5

Author(s):

Yong Li ◽

Dapeng Wu ◽

Pei Wang ◽

Xiaohui Li ◽

Gongning Shi

Keyword(s):

Cell Proliferation ◽

Esophageal Carcinoma ◽

Signaling Pathway ◽

Specific Inhibitor ◽

Inhibitory Effect ◽

Luciferase Reporter ◽

Ki 67 ◽

Proliferation And Apoptosis ◽

Ec Cells ◽

Induced Apoptosis

miR-195 is related to tumorigenesis and frequently inhibits cell proliferation and promotes apoptosis in various cancers, including esophageal carcinoma (EC). The mTOR/p70s6k signaling pathway, which is the major target pathway for HMGA2, regulates the survival and cell proliferation of many tumors and is commonly active in EC. The relationships of miR-195, HMGA2, and the mTOR/p70s6k signaling pathway in EC, however, remain unknown. In the present study, we found that the miR-195 level was significantly downregulated in EC tissues, while the mRNA expressions of HMGA2 were significantly upregulated. Dual-luciferase reporter assay demonstrated that HMGA2 is a target of miR-195. MTT assay and flow cytometry revealed that miR-195 overexpression inhibited cell proliferation and induced apoptosis by targeting HMGA2. We also found that HMGA2 restored the inhibitory effect of miR-195 on phosphorylation of mTOR and p70S6K. Furthermore, rapamycin, a specific inhibitor of the mTOR/p70S6K signaling pathway, decreased the levels of Ki-67 and Bcl-2/Bax ratio, inhibited cell proliferation, and promoted apoptosis in EC cells. In conclusion, upregulation of miR-195 significantly suppressed cell growth and induced apoptosis of EC cells via suppressing the mTOR/p70s6k signaling pathway by targeting HMGA2.

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Immunohistochemical WWOX Expression and Association with Angiogenesis, p53 Expression, Cell Proliferation and Clinicopathological Parameters in Cervical Cancer

Revista Brasileira de Ginecologia e Obstetrícia / RBGO Gynecology and Obstetrics ◽

10.1055/s-0037-1618597 ◽

2018 ◽

Vol 40 (02) ◽

pp. 079-085 ◽

Cited By ~ 3

Author(s):

Mariana Seabra ◽

Eduardo Cândido ◽

Paula Vidigal ◽

Rivia Lamaita ◽

Angélica Rodrigues ◽

...

Keyword(s):

Cell Proliferation ◽

Cervical Squamous Cell Carcinoma ◽

Pelvic Lymphadenectomy ◽

Histological Evaluation ◽

Clinicopathological Parameters ◽

Control Group ◽

Ki 67 ◽

Tissue Samples ◽

P53 Expression ◽

Hematoxylin And Eosin

Objective The current study evaluated the expression of WW domain-containing oxidoreductase (WWOX), its association with clinicopathological features and with p53, Ki-67 (cell proliferation) and CD31 (angiogenesis) expression in patients with invasive cervical squamous cell carcinoma (ICSCC). To the best of our knowledge, no other study has evaluated this association. Methods Women with IB stage-ICSCC (n = 20) and women with uterine leiomyoma (n = 20) were prospectively evaluated. Patients with ICSCC were submitted to type B-C1 radical hysterectomy and pelvic lymphadenectomy. Patients in the control group underwent vaginal hysterectomy. Tissue samples were stained with hematoxylin and eosin for histological evaluation and protein expression was detected by immunohistochemistry studies. Results The WWOX expression was significantly lower in the tumor compared with the expression in the benign cervix (p = 0.019). The WWOX expression was inversely associated with the CD31 expression in the tumor samples (p = 0.018). There was no association between the WWOX expression with the p53 expression (p = 0.464) or the Ki-67 expression (p = 0.360) in the samples of invasive carcinoma of the cervix. There was no association between the WWOX expression and tumor size (p = 0.156), grade of differentiation (p = 0.914), presence of lymphatic vascular invasion (p = 0.155), parametrium involvement (p = 0.421) or pelvic lymph node metastasis (p = 0.310) in ICSCC tissue samples. Conclusion The results suggested that WWOX may be involved in ICSCC carcinogenesis, and this marker was associated with tumor angiogenesis.

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Platelet-Rich Plasma Suppresses Inflammation Reaction of Rat Articular Chondrocytes via Wingless-Related Integration Site/β-Catenin Axis

Journal of Biomaterials and Tissue Engineering ◽

10.1166/jbt.2021.2688 ◽

2021 ◽

Vol 11 (7) ◽

pp. 1372-1376

Author(s):

Wei Wang ◽

Weiwei Ma ◽

Xu Li ◽

Yihui Huang ◽

Xinyu Cao

Keyword(s):

Second Generation ◽

Articular Chondrocytes ◽

Glycogen Synthase ◽

Integration Site ◽

Platelet Rich Plasma ◽

Inflammatory Factors ◽

Blue Staining ◽

Saline Control ◽

Control Group ◽

Mrna Levels

Our study aims to elucidate the role of platelet-rich plasma (PRP) in rats chondrocytes inflammation and mechanism. PRP was obtained from 8 weeks old rats. Then, the knee joint of bilateral hind limbs was dissected and articular chondrocytes were obtained in super-clean table after dislocation and identified at the second generation during culture and passage. Chondrocytes were divided into control group 1 (addition of saline), control group 2 (IWP-2, Wnt/β-catenin axis inhibitor) and experimental group (PRP) followed by analysis of mRNA levels of glycogen synthase kinase-3 (GSK-3β), low-density lipoprotein receptor-associated protein 5 (LRP5), Wnt1 and β-catenin by RT-PCR, IL-1 and TNF-α after 1 week by ELISA. The second generation articular chondrocytes presented polygonal or triangular cell morphology, positive for collagen II and toluidine blue staining. PRP addition significantly reduced GSK-3β and LRP5 mRNA level, and increased β-catenin and Wnt1 mRNA levels in chondrocytes. Meanwhile, it suppressed IL-1 and TNF-α secretion and Wnt protein production inhibitor 2. PRP might suppresses inflammatory factors production of rat articular chondrocytes through inhibiting Wnt/β-catenin axis.

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SLC39A1 Contributes to Gemcitabine Resistance of Pancreatic Ductal Adenocarcinoma by Activating AMPK Signaling

10.21203/rs.3.rs-856017/v1 ◽

2021 ◽

Author(s):

Hao Yu ◽

Xiaoping Mei ◽

Xueming Zhang ◽

Neng Qian ◽

Qingjiang Yu ◽

...

Keyword(s):

Cell Proliferation ◽

Protein Expression ◽

Pancreatic Ductal Adenocarcinoma ◽

Western Blotting ◽

Poor Prognosis ◽

Ductal Adenocarcinoma ◽

Tumor Type ◽

Ampk Signaling ◽

Gemcitabine Resistance ◽

Mrna And Protein Expression

Abstract Objective: Pancreatic ductal adenocarcinoma (PDAC) serves as a prevailing tumor type with high mortality and poor prognosis. The study aims to explore the mechanism of gemcitabine resistance in PDAC patients. Methods: Immunohistochemistry(IHC)was used to analyze the expression of SLC39A1 in PDAC samples. PDAC cells were culture and transfected with siSLC39A1 and siNC, respectively. Cell proliferation analysis was performed using CCK-8 assay. And qPCR and Western blotting was used to analysis the expression level of SLC39A1 and related signal molecular in cells. Results: IHC results demonstrated that the SLC39A1 expression was significantly up-regulated in the gemcitabine-resistant PDAC samples compared with gemcitabine-sensitive PDAC samples. The treatment of gemcitabine dose-dependently inhibited the viability of the PDAC cells. Meanwhile, the mRNA and protein expression of SLC39A1 were elevated in the gemcitabine-resistant PDAC. The treatment of gemcitabine remarkably decreased viability of PDACs, in which SLC39A1 depletion could reverse this effect. SLC39A1 knockdown could reverse the gemcitabine-induced phosphorylation of AMPK enhanced and gemcitabine-inhibited S6K expression. Conclusion: SLC39A1 contributed to gemcitabine resistance of PDAC by activating AMPK signaling.

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Effect of SRY-Related High Mobility Group Box 9 on Extracellular Signal-Regulated Kinase/p38 Signaling Pathway in Glioma

Journal of Biomaterials and Tissue Engineering ◽

10.1166/jbt.2021.2531 ◽

2021 ◽

Vol 11 (1) ◽

pp. 171-175

Author(s):

Tianlong Quan ◽

Chunhua Zhang ◽

Xin Song ◽

Lu Wang

Keyword(s):

Cell Proliferation ◽

Signaling Pathway ◽

Cell Invasion ◽

Cell Apoptosis ◽

Caspase 3 ◽

High Mobility ◽

Negative Control ◽

Glioma Cell Line ◽

High Mobility Group ◽

Control Group

As a common malignant tumor in neurosurgery, glioma is characterized as high incidence rate, easy to invade, metastasize and recurrent. It is difficult to treat and has a poor prognosis. The gliomas pathogenesis is complex and has not been fully resolved. Therefore, finding effective molecular targets for glioma is beneficial to improve therapeutic effect. The SRY-related high mobility group box 9 (SOX9) gene involves in mammalian development and is significantly increased in glioma. However, SOX9’s role in gliomas is unclear. The glioma cell line U87 was assigned into control group, scramble group that was transfected with siRNA negative control, and SOX9 siRNA group that was transfected with SOX9 siRNA followed by analysis of SOX9 mRNA and protein level by qPCR and Western blot, cell proliferation by MTT assay, cell apoptosis by Caspase 3 activity assay, cell invasion by Transwell assay, and MMP-9 level by ELISA. SOX9 siRNA transfection significantly downregulated SOX9 mRNA and protein expressions, inhibited U87 cell proliferation, enhanced Caspase 3 activity, suppressed cell invasion of U87, decreased the secretion of MMP-9 in the supernatant, and reduced ERK1/2 and P38 phosphorylation levels (P < 0.05). SOX9 can regulate the progression of glioma by regulating ERK/P38 signaling pathway, promoting cell apoptosis, inhibiting cell proliferation, and restraining cell invasion.

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