scholarly journals Relationship between IL10 and PD-L1 in Liver Hepatocellular Carcinoma Tissue and Cell Lines

2020 ◽  
Vol 2020 ◽  
pp. 1-14
Author(s):  
Qian Qian ◽  
Changping Wu ◽  
Jianping Chen ◽  
Weibing Wang
Keyword(s):  
Hepatocellular Carcinoma ◽  
Monoclonal Antibody ◽  
Cell Lines ◽  
Western Blotting ◽  
Large Scale ◽  
Expression Level ◽  
Survival Duration ◽  
Great Promise ◽  
Clinical Response Rate ◽  
Liver Hepatocellular Carcinoma

Background. Despite the large-scale clinical application of programmed death-ligand 1 (PD-L1) monoclonal antibody, reduction in its clinical response rate has become a gradual problem. As such, use of PD-L1 monoclonal antibody in combination with other anticarcinoma drugs has been the main strategy in improving its efficacy. Interleukin 10 (IL10) is a recognized inflammatory and immunosuppressive factor. Previous studies have suggested that there is a link between PD-L1 and IL10. Objective. This study was aimed at clarifying the relationship between PD-L1 and IL10 in liver hepatocellular carcinoma (LIHC) and whether IL10 enhances the efficacy of PD-L1 inhibitor. Methods. Expression levels of PD-L1 and IL10 in carcinoma and adjacent tissues were tested by immunochemistry, Western blotting, and RT-PCR. Survival duration and follow-up data of each patient were recorded. LIHC cell lines Bel7405 and MHCC 97-H were used for in vitro experiments. Exogenous IL10 and anti-IL10 were added to cell supernatant. Expression level of PD-L1 in the LIHC cell lines was determined using Western blotting and ELISA. CCK8 and transwell assays were adopted to examine the effect of PD-L1 combined with IL10 on proliferation, invasion, and metastasis of LIHC cells. Results. The survival period of patients with low expression of IL10 was longer than that of patients with high expression (P=0.01). Overexpression of PD-L1 increased the IL10 and Met levels in LIHC tissues and cell lines. IL10 downregulated the expression level of PD-L1 and enhanced the efficacy of crizotinib via the Met signaling pathway in the LIHC cells. Conclusions. A combination of IL10 and PD-L1 inhibitor holds great promise as an effective treatment for LIHC.

Effect of 5'-fluoro-2'-deoxycytidine and sodium butyrate on the genes of the intrinsic apoptotic pathway, p21, p53, cell viability, and apoptosis in human hepatocellular carcinoma cell lines

2021 ◽  
Author(s):  
Masumeh Sanaei ◽  
Fraidoon Kavoosi ◽  
Mohammad Amin Moezzi
Keyword(s):  
Gene Expression ◽  
Hepatocellular Carcinoma ◽  
Cell Viability ◽  
Cell Lines ◽  
Sodium Butyrate ◽  
Cell Apoptosis ◽  
Chromatin Organization ◽  
Expression Level ◽  
Intrinsic Apoptotic Pathway ◽  
Apoptotic Pathway

Backgrounds: Epigenetic regulation such as DNA methylation plays a major role in chromatin organization and gene transcription. Additionally, histone modification is an epigenetic regulator of chromatin structure and influences chromatin organization and gene expression. The relationship between DNA methyltransferase (DNMTs) expression and promoter methylation of the tumor suppressor genes (TSGs) has been reported in various cancers. Previously, the effect of 5-aza-2'-deoxycytidine (5-AZA-CdR), trichostatin A (TSA), and valproic acid (VPA) was shown on various cancers. This study aimed to investigate the effect of 5'-fluoro-2'-deoxycytidine (FdCyd) and sodium butyrate on the genes of the intrinsic apoptotic pathway, p21, p53, cell viability, and apoptosis in human hepatocellular carcinoma SNU449, SNU475, and SNU368 cell lines. Materials and Methods: In this lab trial study, the SNU449, SNU475, and SNU368 cells were cultured and treated with 5'-fluoro-2'-deoxycytidine and sodium butyrate. To determine cell viability, cell apoptosis, and the relative gene expression level, MTT assay, flow cytometry assay, and qRT-PCR were done respectively. Results: 5'-fluoro-2'-deoxycytidine and sodium butyrate changed the expression level of the BAX, BAK, APAF1, Bcl-2, Bcl-xL, p21, and p53 gene (P<0.0001) by which induced cell apoptosis and inhibit cell growth in all three cell lines, SNU449, SNU475, and SNU368.  Conclusion: Both compounds played their roles through the intrinsic apoptotic pathway to induce cell apoptosis.

scholarly journals Exploration of prognostic index based on immune-related genes in patients with liver hepatocellular carcinoma

2020 ◽  
Vol 40 (7) ◽  
Author(s):  
Weidong Shi ◽  
Lanyun Feng ◽  
Shu Dong ◽  
Zhouyu Ning ◽  
Yongqiang Hua ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Function Analysis ◽  
Prognostic Index ◽  
Prognostic Indicator ◽  
Expression Level ◽  
Clinical Feature ◽  
Molecular Characteristics ◽  
Synthetic Index ◽  
Immune Related Genes ◽  
Liver Hepatocellular Carcinoma

Abstract The present study aimed to screen the immune-related genes (IRGs) in patients with liver hepatocellular carcinoma (LIHC) and construct a synthetic index for indicating the prognostic outcomes. The bioinformatic analysis was performed on the data of 374 cancer tissues and 50 normal tissues, which were downloaded from TCGA database. We observed that 17 differentially expressed IRGs were significantly associated with survival in LIHC patients. These LIHC-specific IRGs were validated with function analysis and molecular characteristics. Cox analysis was applied for constructing a RiskScore for predicting the survival. The RiskScore involved six IRGs and corresponding coefficients, which was calculated with the following formula: RiskScore = [Expression level of FABP5 *(0.064)] + [Expression level of TRAF3 * (0.198)] + [Expression level of CSPG5 * (0.416)] + [Expression level of IL17D * (0.197)] + [Expression level of STC2 * (0.036)] + [Expression level of BRD8 * (0.140)]. The RiskScore was positively associated with the poor survival, which was verified with the dataset from ICGC database. Further analysis revealed that the RiskScore was independent of any other clinical feature, while it was linked with the infiltration levels of six types of immune cells. Our study reported the survival-associated IRGs in LIHC and then constructed IRGs-based RiskScore as prognostic indicator for screening patients with high risk of short survival. Both the screened IRGs and IRGs-based RiskScore were clinically significant, which may be informative for promoting the individualized immunotherapy against LIHC.

scholarly journals IMP3 accelerates the progression of prostate cancer through inhibiting PTEN expression in a SMURF1-dependent way

2020 ◽  
Vol 39 (1) ◽  
Author(s):  
Xiang Zhang ◽  
Dawei Wang ◽  
Boke Liu ◽  
Xingwei Jin ◽  
Xianjin Wang ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Cell Viability ◽  
Signaling Pathway ◽  
Cell Lines ◽  
Western Blotting ◽  
Cell Apoptosis ◽  
Expression Level ◽  
Mtor Signaling Pathway ◽  
Expression Levels ◽  
Cancer Tissues

Abstract Background Insulin-like growth factor 2 (IGF2) messenger RNA binding protein 3 (IMP3) has been testified to be overexpressed in prostate cancer and strongly related to patients’ poor prognosis. However, the functions of IMP3 and the underlying mechanisms in prostate cancer still remain unknown. Therefore, the current study was carried out to reveal the role and molecular mechanism of IMP3 in prostate cancer progression. Methods The expression levels of IMP3 in prostate cancer tissues and cells were detected by immunohistochemistry (IHC), western blotting and RT-PCR. CCK-8, clone formation, flow cytometry and in vivo tumor formation assays were used to determine cell growth, clone formation apoptosis and tumorigenesis, respectively. The effect of IMP3 on the expression levels of the key proteins in PI3K/AKT/mTOR signaling pathway, including PIP2, PIP3, p-AKT, AKT, p-mTOR, mTOR, PTEN and BAD activation of was determined by western blotting. IP (Immunoprecipitation) assay was used to evaluate the effects of IMP3 and SMURF1 (SMAD specific E3 ubiquitin protein ligase 1) on the ubiquitination of PTEN protein. Results IMP3 expression level was significantly increased in prostate cancer tissues and cell lines (LNCap, PC3 and DU145) as compared with the paracancerous normal tissues and cells (RWPE-1), respectively. High expression of IMP3 apparently promoted cell viability, tumorigenesis and inhibited cell apoptosis in prostate cancer LNCap, DU145 and PC3 cell lines. In mechanism, IMP3 upregulation significantly increased the phosphorylation levels of AKT and mTOR, and elevated PIP3 expression level, while induced significant reductions in the expression levels of BAD, PTEN and PIP2. And, IMP3 overexpression increased SMURF1 expression, which facilitated PTEN ubiquitination. In addition, SMURF1 overexpression enhanced prostate cancer cell viability and inhibited cell apoptosis. Silence of SMURF1 rescued the enhancements in cell proliferation and tumorigenesis and the inhibition in cell apoptosis rates induced by IMP3 in prostate cancer DU145 and LNCap cells. Conclusion This study reveals that IMP3 is overdressed in prostate cancer, which accelerates the progression of prostate cancer through activating PI3K/AKT/mTOR signaling pathway via increasing SMURF1-mediated PTEN ubiquitination.

scholarly journals Plexin C1 Marks Liver Cancer Cells with Epithelial Phenotype and Is Overexpressed in Hepatocellular Carcinoma

2018 ◽  
Vol 2018 ◽  
pp. 1-9 ◽  
Author(s):  
Gorkem Odabas ◽  
Metin Cetin ◽  
Serdar Turhal ◽  
Huseyin Baloglu ◽  
A. Emre Sayan ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Monoclonal Antibody ◽  
Cell Lines ◽  
Tissue Microarrays ◽  
Mesenchymal Phenotype ◽  
Protein Levels ◽  
Epithelial Phenotype ◽  
Well Differentiated

Background and Aims. Hepatocellular carcinoma is an aggressive malignancy of the liver and is ranked as the sixth most common cancer worldwide. There is still room for novel markers to improve the diagnosis and monitoring of HCC. Our observations in cancer databases that PLXNC1 is upregulated in HCC led us to investigate the expression profile of Plexin C1 mRNA and protein in HCC cell lines and tissues. Methods. A recombinant protein encompassing part of the extracellular domain of Plexin C1 was used as an antigen for monoclonal antibody development. Transcript and protein levels of Plexin C1 in HCC cell lines were determined by RT-qPCR and Western blotting, respectively. In vivo evaluation of Plexin C1 expression in HCC tissues was accomplished by immunohistochemistry studies in tissue microarrays. Results. A monoclonal antibody, clone PE4, specific to Plexin C1, was generated. In silico and in vitro analyses revealed a Plexin C1-based clustering of well-differentiated HCC cell lines. Staining of HCC and nontumoral liver tissues with PE4 showed a membrane-localized overexpression of Plexin C1 in tumors (p=0.0118). In addition, this expression was correlated with the histological grades of HCC cases. Conclusions. Plexin C1 distinguishes HCC cells of epithelial characteristics from those with the mesenchymal phenotype. Compared to the nontumoral liver, HCC tissues significantly overexpress Plexin C1. The newly generated PE4 antibody can be evaluated in larger HCC cohorts and might be exploited for the examination of Plexin C1 expression pattern in other epithelial malignancies.

scholarly journals Altered staining patterns and expression level of Engrailed-2 in Benign prostatic hyperplasia and Prostate Cancer predict Prostatic disease progression

2020 ◽  
Author(s):  
Qi Li ◽  
Yibo Shi ◽  
Rigai Sa ◽  
Jun Hao ◽  
Jinhao Hu ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Monoclonal Antibody ◽  
Benign Prostatic Hyperplasia ◽  
Disease Progression ◽  
Cell Lines ◽  
Immunohistochemical Staining ◽  
Prostatic Hyperplasia ◽  
Expression Level ◽  
Clinical Staging ◽  
Rt Pcr

Abstract Background: Prostate cancer (PC) , a common malignant tumor, is the second-leading cause of cancer death among American men. Its successful treatment greatly relies on the early diagnose. Engrailed-2 (EN2) has been confirmed being existed with a high level in the urine of PC patients. In this study, to explore the application of EN2 in PC, we detected the immunohistochemical staining difference and EN2 expression level between benign prostatic hyperplasia (BPH) and PC. Methods: We developed a monoclonal antibody against the helix 3 in EN2 and confirmed its specificity with Western blotting (WB) and immunofluorescence detecting the subcellular localization of endogenous and exogenous EN2 in three PC cell lines (LNCap, PC3, and DU145). We conducted immunohistochemical staining using this homemade antibody, and RT-PCR to detect the expression of EN2 in 25 PC and 25 BPH cases , and analyzed the correlation of EN2 expression and PC clinical staging. Results: The results of WB and immunofluorescence showed our homemade EN2 monoclonal antibody could specifically bind endogenous and exogenous EN2 protein in three different PC cell lines. Endogenous EN2 was generally expressed in the cytoplasm and exogenous EN2 mostly existed in the nucleus of these cell lines. Immunohistochemical staining in PC had extremely stronger signals than that in BPH, suggesting a higher EN2 expression level in PC, which was confirmed by RT-PCR. Interestingly, the stained areas in BPH tissues were mainly in nucleus and cytoplasm, while in PC tissues were mainly on cytomembrane. Moreover, the expression level of EN2 was positively correlated with the PC clinical staging. Conclusion: Using our homemade EN2 antibody, we have found different staining patterns and expression level of EN2 in BPH and PC，which may be helpful to predict prostatic disease progression.

scholarly journals Relationship between IL-10 and PD-L1 in liver hepatocellular carcinoma tissue and cell lines

2020 ◽  
Author(s):  
Qian Qian ◽  
Changping Wu ◽  
Jianping Chen ◽  
Weibing Wang
Keyword(s):  
Hepatocellular Carcinoma ◽  
Cell Lines ◽  
Cancer Progression ◽  
Critical Role ◽  
Clinical Importance ◽  
Interleukin 10 ◽  
Migration And Invasion ◽  
Western Blots ◽  
Expression Levels ◽  
Liver Hepatocellular Carcinoma

Abstract Background Programmed death-ligand1 (PD-L1) plays a critical role in host immunity in the setting of cancer progression. Interleukin 10 (IL-10) is a multi-cellular, multi-functional cytokine that regulates cell growth and differentiation and participates in inflammatory and immune responses. The purpose of this study was to clarify the relationship between PD-L1 and IL-10 and their clinical importance in liver hepatocellular carcinoma(LIHC).Methods LIHC patients (n=100) who underwent surgery with preoperative therapy were included in the study. By immuno-histochemical staining, PD-L1, IL-10 and CD8 positive cells were examined in resected specimens. The gene expression levels of PD-L1, IL-10 and Met were detected by qRT-PCR and Western blots, and differentially compared in cancer, adjacent and normal tissues. In cell experiments, the Bel7405 and MHCC97-H cell-lines were incubated with IL-10 or anti-IL-10 antibody, and then PD-L1 and Met expression levels were compared by ELISA and Western blots. The effect of crizotinib and/or IL-10 on the proliferation, invasion and migration of LIHC cell-lines was estimated by CCK8 and transwell assay.Results In tumor tissues, the mRNA and protein levels of PD-L1, IL-10 and Met were higher than those in adjacent tissues. The high expression levels of PD-L1 and IL-10 indicated a poor prognosis. IL-10 reduced the expression of PD-L1 in LIHC cell-lines via Met signaling pathway. Over-expression of PD-L1 in increased the levels of IL-10, and Met in in LIHC tissue and cell lines. The combination of crizotinib and IL-10 were more effective in inhibiting the proliferation, migration and invasion of LIHC cell lines. Conclusions The combination of IL-10 and PD-L1 monoclonal antibody may have therapeutic promise in treating LIHC.

scholarly journals Antiproliferative Activity Evaluation of a Series ofN-1,3-Benzothiazol-2-ylbenzamides as Novel Apoptosis Inducers

2016 ◽  
Vol 2016 ◽  
pp. 1-5 ◽  
Author(s):  
Filomena Corbo ◽  
Alessia Carocci ◽  
Domenico Armenise ◽  
Nicolino De Laurentis ◽  
Antonio Laghezza ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Hepatocellular Carcinoma ◽  
Cell Lines ◽  
Antiproliferative Activity ◽  
Human Breast Cancer ◽  
Human Liver ◽  
Inhibitory Effect ◽  
Human Breast ◽  
Liver Hepatocellular Carcinoma ◽  
Mcf 7

A series ofN-1,3-benzothiazol-2-ylbenzamide derivatives were studied for their antiproliferative activity on human liver hepatocellular carcinoma (HepG2) and human breast cancer (MCF-7) cell lines. Most of them were found to show a prominent inhibitory effect on cell growth. Among the most active compounds,1kemerged for its proapoptotic effect that is particularly evident towards MCF-7 cancer cell lines.

scholarly journals Clinicopathologic Significance of G Protein-Coupled Receptor 81 in Hepatocellular Carcinoma

2017 ◽  
Vol 2 (2) ◽  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Prognostic Factor ◽  
Cell Line ◽  
G Protein ◽  
Cell Lines ◽  
Expression Level ◽  
Migration And Invasion ◽  
G Protein Coupled Receptor ◽  
Normal Liver Cell ◽  
G Protein Coupled

Background/ Aims: Lactate functions as a metabolic key player in cancer in various aspects. G-protein coupled receptor 81 (GPR81), a cell surface lactate receptor, is involved in the metabolism of lactate. However, only a few studies have been conducted on GPR81 expression in cancer, especially hepatocellular carcinoma (HCC). The present study aims to identify the clinical significance of GPR81 expression in HCC and its role as a prognostic factor. Methods: Tissues were obtained from 197 patients who had undergone surgery for HCC. GPR81 expression level was assessed by immunohistochemistry. And the function of GPR81 on HCC cell growth and mobility was explored through cell line studies. Results: GPR81 was overexpressed in the HepG2, Huh7, SNU182 and SK-Hep1 HCC cell lines and HCC tissues compared with that in the THLE-2 normal liver cell lines. Furthermore, high GPR81 expression levels were correlated significantly with disease recurrence. In addition, because of significant differences in cancer proliferation, migration, and invasion depending on the GPR81 expression level in various HCC cell line studies, GPR81 is believed to play a role in promoting aggressive cancer cell behavior. Conclusions: As such, GPR81 expression level was determined to be a useful prognostic factor for predicting HCC progression. The present study is the first to report on GPR81 expression in HCC and its significance. Henceforth, GPR81 is expected to become a highly valuable candidate for future target therapy.

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