scholarly journals Integrative Analysis of HMMR as a Potential Target of Prognosis and Therapy in Hepatocellular Carcinoma

2021 ◽  
pp. 1-12
Author(s):  
Yilan Huang ◽  
Bin Yu ◽  
Qinhui Liu ◽  
Xuping Yang ◽  
Xin Liu ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Overall Survival ◽  
Western Blotting ◽  
Gene Set Enrichment Analysis ◽  
Rt Pcr ◽  
Poor Prognostic Factor ◽  
Western Blotting Analysis ◽  
Blotting Analysis ◽  
Potential Target ◽  
Tcga Dataset

Background: Previous work has indicated Hyaluronic acid-mediated motor receptor (HMMR) plays an important role in regulating tumor metastasis. However, few researchers address the clinical significance of HMMR and its underlying mechanisms for regulating hepatocellular carcinoma (HCC). This study focuses on the underlying effect of HMMR in the development and prognosis of HCC. Materials and Methods: In the present study, data of RNA and miRNA sequencing array were obtained from Oncomine dataset or The Cancer Genome Atlas (TCGA) dataset, the distinctive genomic patterns associated with HMMR expression and its correlation with prognosis were analysed by using R package. Gene set enrichment analysis (GSEA) were performed on genes expressed aberrantly. We also performed Reverse Transcription-polymerase Chain Reaction (RT-PCR), Immunohistochemical (IHC) staining and Western blotting analysis to evaluate the expression of HMMR in liver cancer cell lines or 12 HCC samples from The Affiliated Hospital of Southwest Medical University. Results: A total of 407 tumor tissue samples in TCGA dataset were evaluated, combined with analysis in Oncomine dataset, we found HMMR expression was increased in HCC compared to normal tissues. Higher expression of HMMR was correlated with poorer overall survival and disease-free survival outcomes. Moreover, multivariate Cox regression analysis revealed that HMMR expression was an independent risk factor for overall survival (HMMR: hazard ratio [HR] = 1.154, 95% confidence interval [CI] = 1.080-1.233, p<0.001). Consistently, RT-PCR, IHC staining and Western blotting analysis further confirmed that HMMR expression was increased in HCC compared with patient-matched adjacent normal liver tissues. Notably, GSEA analysis revealed that differential gene expression in HMMR-high patients (compared with HMMRlow patients) were enriched in cell proliferation and p53 signaling pathway. Moreover, comprehensive analysis showed a strong correlation between HMMR upregulation and miRNA changes. Conclusion: The high expression of HMMR is a poor prognostic factor in HCC and might serve as a potential target of therapy in patients with HCC.

scholarly journals Reduced Expression of Corticotropin-Releasing Hormone Receptor Type-1α in Human Preeclamptic and Growth-Restricted Placentas

2003 ◽  
Vol 88 (1) ◽  
pp. 363-370 ◽  
Author(s):  
E. Karteris ◽  
A. Goumenou ◽  
E. Koumantakis ◽  
E. W. Hillhouse ◽  
D. K. Grammatopoulos
Keyword(s):  
Western Blotting ◽  
Dynamic Balance ◽  
Maternal Plasma ◽  
Receptor Expression ◽  
Control Group ◽  
Mrna Levels ◽  
Angiotensin Ii Receptor ◽  
Rt Pcr ◽  
Western Blotting Analysis ◽  
Blotting Analysis

Placentally derived CRH seems to play a major role in the mechanisms controlling human pregnancy and parturition, via activation of specific receptors widespread in reproductive tissues. In the human placenta, CRH seems to modulate vasodilation, prostaglandin production, and ACTH secretion. It has also been suggested that CRH might act as a placental clock, determining the length of gestation. In addition, maternal plasma CRH concentrations are further elevated in pregnancies associated with abnormal placental function, such as preeclampsia and intrauterine growth retardation (IUGR). In this study, we sought to investigate the expression of CRH-R1α levels in placentas from women who have undergone normal deliveries (control group) and patients who have been diagnosed as having preeclampsia or IUGR. Results showed that placental CRH-R1α mRNA levels (as shown by quantitative RT-PCR) and protein levels (shown by Western blotting analysis) were significantly (P &lt; 0.05) reduced in all of the complicated pregnancies. In contrast, levels of the angiotensin II receptor were elevated in preeclampsia and reduced in IUGR subjects, as shown by RT-PCR and Western blotting analysis. These findings might suggest that changes in receptor expression may contribute toward dysregulation of the dynamic balance controlling vascular resistance.

scholarly journals Analysis of the physical activity effects and measurement of pro-inflammatory cytokines in irradiated lungs in rats

2012 ◽  
Vol 27 (3) ◽  
pp. 223-230 ◽  
Author(s):  
Renata Cristiane Gennari Bianchi ◽  
Eduardo Rochete Ropelle ◽  
Carlos Kiyoshi Katashima ◽  
José Barreto Campello Carvalheira ◽  
Luiz Roberto Lopes ◽  
...  
Keyword(s):  
Physical Activity ◽  
Western Blotting ◽  
Inflammatory Cytokines ◽  
Lower Lobe ◽  
Whole Body ◽  
Control Group ◽  
Western Blotting Analysis ◽  
Blotting Analysis ◽  
Tnf Α ◽  
Pro Inflammatory Cytokines

PURPOSE: To study if the pre-radiotherapy physical activity has radio-protective elements, by measuring the radio-induced activation of pro-inflammatory cytokines as interleukin-6 (il-6), transforming growth factor -β (tgf -β), tumor necrosis factor -α (tnf-α) and protein beta kinase β (ikkβ), through western blotting analysis. METHODS: A randomized study with 28 Wistar hannover rats, males, with a mean age of 90 days and weighing about 200 grams. The animals were divided into three groups: (GI, GII and GIII). GIII group were submitted to swimming for eight weeks (zero load, three times a week, about 30 minutes). Then, the groups (except the control group) were submitted to irradiation by cobalt therapy, single dose of 3.5 gray in the whole body. All animals were sacrificed by overdose of pentobarbital, according to the time for analysis of cytokines, and then a fragment of the lower lobe of the right lung went to western blotting analysis. RESULTS: The cytokines IKK β, TNF-α and IL-6 induced by radiation in the lung were lower in the exercised animals. However, exercise did not alter the radiation-induced increase in tgf-β. CONCLUSION: The results show a lower response in relation to inflammatory cytokines in the group that practiced the exercise pre-radiotherapy, showing that exercise can protect tissues from tissue damage due to irradiation.

scholarly journals Mechanisms Involved in the Anti-Tumor Effects of Toosendanin in Glioma Cells

2021 ◽  
Author(s):  
haiyan huang ◽  
Chaochao Zhang ◽  
Haijun Gao ◽  
Ziqiang Liu ◽  
Jiacheng Lai ◽  
...  
Keyword(s):  
Flow Cytometry ◽  
Wound Healing ◽  
Western Blotting ◽  
Colony Formation ◽  
Glioma Cell ◽  
Glioma Cells ◽  
Migration And Invasion ◽  
Antitumor Effects ◽  
Western Blotting Analysis ◽  
Blotting Analysis

Abstract Background: Toosendanin (TSN) is a triterpenoid compound mainly used as an ascaris repellant. Recent studies have shown that it possesses antitumor effects in many types of tumor cells. However, the effects of TSN on glioma cells have rarely been reported. Methods: Different assays were performed to investigate the effects of TSN on the different glioma cell lines including U87MG and LN18. The assays included colony formation, wound healing, and transwell assays. Furthermore, Hoechst 3342 staining, flow cytometry, and western blotting analysis were performed to investigate the apoptotic activities of TSN. Finally, the results were confirmed using a xenograft tumor model that comprised of nude mice. Results: In vitro, the CCK-8 and colony formation assays showed that TSN effectively inhibited glioma cell proliferation. Moreover, the inhibitory effects on glioma cell migration and invasion were demonstrated through the wound healing and transwell assays, respectively. Hoechst 33342 staining, flow cytometry, and western blotting assays demonstrated the significant effect of TSN in the apoptosis induction of glioma cells. Furthermore, the anti-glioma effect of TSN was exerted through the inhibition of the PI3K/Akt/mTOR signaling pathways as demonstrated by western blotting analysis. In addition, the effects of TSN on glioma cell viability, apoptosis, cell cycle arrest, migration, and invasion were reversed by 740Y-P, a PI3K activator. Finally, the mouse xenograft model confirmed the suppressive effect of TSN on tumor growth in vivo. Conclusion: Our results suggest that TSN is a promising chemotherapeutic drug for patients with glioma.

scholarly journals Identification of a five-gene signature in association with overall survival for hepatocellular carcinoma

PeerJ ◽  
2021 ◽  
Vol 9 ◽  
pp. e11273
Author(s):  
Lei Yang ◽  
Weilong Yin ◽  
Xuechen Liu ◽  
Fangcun Li ◽  
Li Ma ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Cell Cycle ◽  
Overall Survival ◽  
Cox Regression ◽  
Enrichment Analysis ◽  
Gene Signature ◽  
Gene Set Enrichment Analysis ◽  
Enrichment Score ◽  
Hub Genes ◽  
Cox Regression Analysis

Background Hepatocellular carcinoma (HCC) is considered to be a malignant tumor with a high incidence and a high mortality. Accurate prognostic models are urgently needed. The present study was aimed at screening the critical genes for prognosis of HCC. Methods The GSE25097, GSE14520, GSE36376 and GSE76427 datasets were obtained from Gene Expression Omnibus (GEO). We used GEO2R to screen differentially expressed genes (DEGs). A protein-protein interaction network of the DEGs was constructed by Cytoscape in order to find hub genes by module analysis. The Metascape was performed to discover biological functions and pathway enrichment of DEGs. MCODE components were calculated to construct a module complex of DEGs. Then, gene set enrichment analysis (GSEA) was used for gene enrichment analysis. ONCOMINE was employed to assess the mRNA expression levels of key genes in HCC, and the survival analysis was conducted using the array from The Cancer Genome Atlas (TCGA) of HCC. Then, the LASSO Cox regression model was performed to establish and identify the prognostic gene signature. We validated the prognostic value of the gene signature in the TCGA cohort. Results We screened out 10 hub genes which were all up-regulated in HCC tissue. They mainly enrich in mitotic cell cycle process. The GSEA results showed that these data sets had good enrichment score and significance in the cell cycle pathway. Each candidate gene may be an indicator of prognostic factors in the development of HCC. However, hub genes expression was weekly associated with overall survival in HCC patients. LASSO Cox regression analysis validated a five-gene signature (including CDC20, CCNB2, NCAPG, ASPM and NUSAP1). These results suggest that five-gene signature model may provide clues for clinical prognostic biomarker of HCC.

scholarly journals Decreased Expression of KPNA2 in Hepatocellular Carcinoma Infers Favorable Prognosis and High Immune Infiltrating Level

2021 ◽  
Author(s):  
kangming zhu ◽  
yvndi zhang ◽  
hui yvan ◽  
jing li
Keyword(s):  
Hepatocellular Carcinoma ◽  
Overall Survival ◽  
Logistic Regression ◽  
Survival Rate ◽  
Western Blotting ◽  
Good Prognosis ◽  
Overall Survival Rate ◽  
Pathological Stage ◽  
Down Regulation ◽  
Immune Infiltration

Abstract BackgroundLiver hepatocellular carcinoma (LIHC) is an important pathological type of liver cancer. The immune infiltration of the tumor microenvironment is negatively correlated with the overall survival rate of LIHC. At present , the role and molecular mechanism of KPNA2 in LIHC have not been elucidated, and the prognostic correlation between the two and the immune infiltration of LIHC are still unclear. Our study evaluated the role of KPNA2 in LIHC through TCGA data.MethodGene expression profiling interactive analysis (GEPIA) is used to analyze the expression of KPNA2 in LIHC. We evaluated the impact of KPNA2 on the survival of LIHC patients through the survival module. Then, We downloaded the LIHC data set from TCGA. Logistic regression was used to analyze the correlation between clinical information and KPNA2 expression. Cox regression analysis was used to analyze the clinicopathological characteristics related to the overall survival rate of TCGA patients. In addition, we used the "correlation" modules of CIBERSORT and GEPIA to explore the correlation between KPNA2 and cancer immune infiltrate. Western blotting was used to detect the expression of KPNA2.ResultUsing logistic regression for univariate analysis, increased KPNA2 expression was significantly correlated with pathological stage, tumor status, and lymph node status. In addition, multivariate analysis showed that down-regulation of KPNA2 expression, negative pathological stage and distant metastasis are independent prognostic factors for good prognosis. Specifically, CIBERSORT analysis was used to establish a negative correlation between the up-regulated expression of KPNA2 and the level of immune infiltration of B cells, NK cells, mast cells, and T cells. In addition, we confirmed this in the "Association" module of GEPIA. The expression of KPNA2 in LIHC tissues was significantly lower than that in adjacent normal tissues by western blotting.ConclusionThe down-regulation of KPNA2 expression is associated with a good prognosis and an increase in the proportion of immune cells in LIHC. These conclusions indicate that KPNA2 is related to the level of immune infiltration of LIHC and can be used as a potential prognostic biomarker of LIHC and a potential target for clinical tumor treatment.

scholarly journals So-Ochim-Tang-Gamibang, a Traditional Herbal Formula, Ameliorates Depression by Regulating Hyperactive Glucocorticoid Signaling In Vitro and In Vivo

2020 ◽  
Vol 2020 ◽  
pp. 1-10
Author(s):  
Mirim Jin ◽  
Sun Young Park ◽  
Hye Jin Choi ◽  
Younmin Shin ◽  
Eunho Chun ◽  
...  
Keyword(s):  
Western Blotting ◽  
Hpa Axis ◽  
Plasma Levels ◽  
Molecular Mechanisms ◽  
Western Blotting Analysis ◽  
Blotting Analysis ◽  
Wky Rats ◽  
Glucocorticoid Signaling

So-ochim-tang-gamibang (SOCG) is a Korean traditional medicine; it has previously been shown to be safe and effective against depression. Persistently increased levels of circulating glucocorticoids have been considered as a pathological mechanism for depression and associated with decreased neurotrophic factors in the hippocampus. This study investigated whether SOCG controls the hyperactivity of the hypothalamic-pituitary-adrenal (HPA) axis and the molecular mechanisms underlying its effects in vivo and in vitro. Wistar Kyoto (WKY) rats were subjected to restraint stress, where SOCG was orally administered to the animals for 2 weeks. An open field test (OFT), forced swimming test (FST), and sucrose preference test (SPT) were performed to explore the antidepressant activity of SOCG in WKY rats. Plasma levels of HPA axis hormones were measured by ELISA or western blotting analysis. The expression levels or activation of HPA axis-related signaling molecules such as brain-derived neurotrophic factor (BDNF), cAMP response element-binding protein (CREB), extracellular regulated kinase (ERK), and glucocorticoid receptors (GRs) in the brain were determined by real-time PCR and western blotting analysis. Furthermore, a corticosterone- (CORT-) induced cell injury model was established using SH-SY5Y cells to explore the antidepressive effects of SOCG in vitro. The results of the OFT, FST, and SPT revealed that SOCG ameliorated depressive-like behaviors in the WKY rats. The blood plasma levels of HPA axis hormones such as CORT, CORT-releasing hormone (CRH), and adrenocorticotrophic hormone were downregulated by SOCG. On the other hand, SOCG upregulated the phosphorylation of CREB and ERK in both the rat hippocampus and CORT-treated SH-SY5Y cells. Moreover, it also increased the GR expression. These results suggested that SOCG may improve depression by controlling hyperactive glucocorticoid signaling via the downregulation of HPA axis hormones and upregulation of GR.

scholarly journals Effects of topical mechanical stability on the formation of Masquelet membrane in a rabbit radial defect model

2020 ◽  
Vol 10 (1) ◽  
Author(s):  
Jie Xie ◽  
Donghao Liu ◽  
Haoyi Wang ◽  
Haitao Long ◽  
Yong Zhu ◽  
...  
Keyword(s):  
Western Blotting ◽  
Mechanical Stability ◽  
Mechanical Test ◽  
Control Group ◽  
Rigid Fixation ◽  
X Ray ◽  
Western Blotting Analysis ◽  
Blotting Analysis ◽  
Fixation Group ◽  
Masquelet Technique

Abstract The exact mechanism of Masquelet technique is unknown. This study intends to explore the effects of topical mechanical stability on the formation of Masquelet membrane. Segmental radius shaft defect was created in all rabbits, which were filled with polymethylmethacrylate (PMMA) in Non-fixation group, and with PMMA fixed with plates in Fixation group, and subjected to no disposal in control group. The topical stability of PMMA and plates were monitored via X-ray and mechanical test. And the membranes were excised for further Histological, IHC and Western-Blotting analysis 4 and 6 weeks post-operatively. X-ray revealed no sign of plates loosening, or shift of PMMA. Mechanical tests revealed superior topical stability by plates. Pathological examinations suggested that vascularized and osteogenic membranes were formed around PMMA. IHC and Western-Blotting analysis revealed that both Fixation and Non-fixation group exerted significant effects on the expression of Ki67, COL I, and CD31 positive cells, as well as the protein expression of osteogenic (RUNX2, ALP) and angiogenic (VEGFA, TGF-β1) factors. And compared with membrane in Non-fixation group, Fixing PMMA spacer with plates caused a significant increase in osteogenic and angiogenic expression. This study indicates that rigid fixation provided by plate in Masquelet technique positively alters the quality of membrane formed surrounding PMMA, in terms of significantly osteogenic and angiogenic potential.

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