Dantrolene Potentiates the Antineoplastic Effect of Sorafenib in Hepatocellular Carcinoma via Targeting Ca+2/PI3K Signaling Pathway

Background: Hepatocellular carcinoma (HCC) is the 6th prevalent cancer and the 4th leading cause of cancer related deaths all over the world. A major challenge for sorafenib, the standard chemotherapeutic agent in HCC treatment, is the chemo-resistance. Objective: This study was conducted to evaluate the role of dantrolene as a possible antineoplastic agent in HCC, and in chemo-sensitization of sorafenib via targeting Ca+2/PI3K pathway. Methods: HCC was induced in rats using a single dose of diethyl nitrosamine (DENA) (200 mg/kg, ip), followed by phenobarbital sodium (0.05%) in drinking water for 18 weeks. At the end of 18th week, rats were allocated into 4 groups (10 rats/each), one group was left without treatment (DENA group) and the other three groups were treated with either sorafenib, dantrolene, or their combination for further 4 weeks. One day after last injection, serum and liver tissues were collected. Liver tissue p53, VEGF, MMP-9, Cyclin D1, PI3K, and, serum AFP were assessed using immunoassay. Hepatic and serum Ca+2 were also computed. Furthermore, Ki67 was assessed immunohistochemically. Results: Dantrolene exhibited synergistic effect when used in combination with sorafenib compared to either drug alone (p <0.05) through decreasing p53, VEGF, MMP-9, Cyclin D1, and Ki-67. In addition, dantrolene was evidenced to have an inhibitory effect on Ca+2/PI3K pathway that mediates its antineoplastic action when used alone or in combination with sorafenib. Conclusion: Dantrolene exerted antineoplastic effect as well as augmented sorafenib antineoplastic activity via intervention of Ca+2/PI3K pathway, manifested by ameliorating angiogenesis, apoptosis, proliferation and metastasis.

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Possible Role of IRS-4 in the Origin of Multifocal Hepatocellular Carcinoma

Cancers ◽

10.3390/cancers13112560 ◽

2021 ◽

Vol 13 (11) ◽

pp. 2560

Author(s):

Luis G. Guijarro ◽

Patricia Sanmartin-Salinas ◽

Eva Pérez-Cuevas ◽

M. Val Toledo-Lobo ◽

Jorge Monserrat ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Liver Cancer ◽

Hepg2 Cells ◽

Cellular Polarity ◽

Ki 67 ◽

Tissue Samples ◽

Vitro Analysis ◽

Tumoral Cells

New evidence suggests that insulin receptor substrate 4 (IRS-4) may play an important role in the promotion of tumoral growth. In this investigation, we have evaluated the role of IRS-4 in a pilot study performed on patients with liver cancer. We used immunohistochemistry to examine IRS-4 expression in biopsies of tumoral tissue from a cohort of 31 patient suffering of hepatocellular carcinoma (HCC). We simultaneously analyzed the expression of the cancer biomarkers PCNA, Ki-67, and pH3 in the same tissue samples. The in vitro analysis was conducted by studying the behavior of HepG2 cells following IRS-4 overexpression/silencing. IRS-4 was expressed mainly in the nuclei of tumoral cells from HCC patients. In contrast, in healthy cells involved in portal triads, canaliculi, and parenchymal tissue, IRS-4 was observed in the cytosol and the membrane. Nuclear IRS-4 in the tumoral region was found in 69.9 ± 3.2%, whereas in the surrounding healthy hepatocytes, nuclear IRS-4 was rarely observed. The percentage of tumoral cells that exhibited nuclear PCNA and Ki-67 were 52.1 ± 7%, 6.1 ± 1.1% and 1.3 ± 0.2%, respectively. Furthermore, we observed a significant positive linear correlation between nuclear IRS-4 and PCNA (r = 0.989; p < 0.001). However, when we correlated the nuclear expression of IRS-4 and Ki-67, we observed a significant positive curvilinear correlation (r = 0.758; p < 0.010). This allowed us to define two populations, (IRS-4 + Ki-67 ≤ 69%) and (IRS-4 + Ki-67 > 70%). The population with lower levels of IRS-4 and Ki-67 had a higher risk of suffering from multifocal liver cancer (OR = 16.66; CI = 1.68–164.8 (95%); p < 0.05). Immunoblot analyses showed that IRS-4 in normal human liver biopsies was lower than in HepG2, Huh7, and Chang cells. Treatment of HepG2 with IGF-1 and EGF induced IRS-4 translocation to the nucleus. Regulation of IRS-4 levels via HepG2 transfection experiments revealed the protein’s role in proliferation, cell migration, and cell-collagen adhesion. Nuclear IRS-4 is increased in the tumoral region of HCC. IRS-4 and Ki-67 levels are significantly correlated with the presence of multifocal HCC. Moreover, upregulation of IRS-4 in HepG2 cells induced proliferation by a β-catenin/Rb/cyclin D mechanism, whereas downregulation of IRS-4 caused a loss in cellular polarity and in its adherence to collagen as well as a gain in migratory and invasive capacities, probably via an integrin α2 and focal adhesion cascade (FAK) mechanism.

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The Role of miRNAs in PI3K Signaling Pathway in Blood Malignancies: A Review Article

Journal of Zabol Medical School ◽

10.18502/jzms.v4i1.6760 ◽

2021 ◽

Author(s):

Haniyeh Gaffari-Nazari ◽

Samira Karami ◽

Leila Noorazar ◽

Sayeh Parkhideh ◽

Elham Roshandel ◽

...

Keyword(s):

Treatment Failure ◽

Signaling Pathway ◽

Intracellular Signaling ◽

Pi3k Pathway ◽

Pi3k Signaling ◽

Laboratory Findings ◽

A Cell ◽

Pi3k Signaling Pathway ◽

Relationship Of

Background: The PI3K/Akt/mTOR signaling pathway is one of the most important intracellular signaling pathways by regulating the cell cycle process. The direct relationship of this pathway with important mechanisms such as cell quiescence, longevity, and proliferation has been established. The overactive PI3K pathway with decreased and increased apoptosis and cell proliferation respectively is involved in pathogenesis of many cancers, including blood malignancies such as leukemia. Methods: Laboratory findings have shown that different factors, such as miRNAs, play a role in regulating PI3K signaling pathway. These molecules can alter the fate of a cell by interfering in suppression/overexpression of mRNA, transcription factors or stimulating the transcription of some genes. In this article, we reviewed the role of miRNAs in regulating the PI3K/Akt/mTOR pathway and its effect on leukemic progression and treatment failure. Conclusion: At present, miRNAs are known to be one of the causes of treatment failure and relapse in cancers.

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The emerging role of the molecular marker p27 in the differential diagnosis of adrenocortical tumors

Endocrine Connections ◽

10.1530/ec-13-0025 ◽

2013 ◽

Vol 2 (3) ◽

pp. 137-145 ◽

Cited By ~ 12

Author(s):

Sofia S Pereira ◽

Tiago Morais ◽

Madalena M Costa ◽

Mariana P Monteiro ◽

Duarte Pignatelli

Keyword(s):

Differential Diagnosis ◽

Cyclin D1 ◽

Adrenal Glands ◽

Malignant Tumors ◽

Molecular Profile ◽

Benign Tumors ◽

Ki 67 ◽

Adrenocortical Tumors ◽

Percentage Area

Malignant adrenocortical tumors (ACTs) are rare and highly aggressive; conversely, benign tumors are common and frequently found incidentally (the so-called incidentalomas). Currently, the use of molecular markers in the diagnosis of ACTs is still controversial. The aim of this study was to analyze the molecular profile of different ACTs with the purpose of identifying markers useful for differentiating between these tumors. The ACTs that were studied (n=31) included nonfunctioning adenomas (ACAn)/incidentalomas (n=13), functioning adenomas with Cushing's syndrome (ACAc) (n=7), and carcinomas (n=11); normal adrenal glands (n=12) were used as controls. For each sample, the percentage area stained for the markers StAR, IGF2, IGF1R, p53, MDM2, p21, p27, cyclin D1, Ki-67, β-catenin, and E-cadherin was quantified using a morphometric computerized tool. IGF2, p27, cyclin D1, and Ki-67 were the markers for which the percentage of stained area was significantly higher in carcinoma samples than in adenoma samples. Ki-67 and p27 were the markers that exhibited the highest discriminative power for differential diagnosis between carcinomas and all type of adenomas, while IGF2 and StAR were only found to be useful for differentiating between carcinomas and ACAn and between carcinomas and ACAc respectively. The usefulness of Ki-67 has been recognized before in the differential diagnosis of malignant tumors. The additional use of p27 as an elective marker to distinguish benign ACTs from malignant ACTs should be considered.

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Long non-coding RNA ZEB1-AS1 promotes proliferation and metastasis of hepatocellular carcinoma cells by targeting miR-299-3p/E2F1 axis

The Journal of Biochemistry ◽

10.1093/jb/mvab042 ◽

2021 ◽

Author(s):

Baiyin Mu ◽

Chenlan Lv ◽

Qingli Liu ◽

Hong Yang

Keyword(s):

Hepatocellular Carcinoma ◽

Cell Proliferation ◽

Luciferase Reporter ◽

Migration And Invasion ◽

Non Coding Rna ◽

Tumorigenesis And Progression ◽

Novel Biomarkers ◽

Proliferation And Metastasis ◽

Long Non Coding Rna

Abstract There is emerging evidence that dysregulation of long non-coding RNAs (lncRNAs) is associated with hepatocellular carcinoma (HCC). Zinc finger E-box binding homeobox 1 antisense 1 (ZEB1-AS1) functions as an oncogenic regulator in various malignancies. Nonetheless, the potential role of ZEB1-AS1 in HCC remains poorly elucidated. Herein, qRT-PCR was employed for examining ZEB1-AS1, miR-299-3p and E2F1 mRNA expressions in HCC cells and tissues. MTT assay was performed to evaluate cell proliferation. Transwell assay was utilized for evaluating cancer cell migration and invasion. Western blot was employed for measuring E2F1 protein expression. What’s more, dual-luciferase reporter assay was utilized for verifying the targeting relationships between ZEB1-AS1 and miR-299-3p, as well as E2F1 and miR-299-3p. It was demonstrated that, in HCC tissues and cells, ZEB1-AS1 expression was markedly increased, and meanwhile, its high expression level is related to the unfavorable clinicopathologic indicators. ZEB1-AS1 overexpression facilitated HCC cell proliferation, migration and invasion, while its knockdown led to the opposite effects. In terms of mechanism, we discovered that ZEB1-AS1 could decoy miR-299-3p and up-regulate E2F1 expression. This work reveals the functions and mechanism of ZEB1-AS1 in HCC tumorigenesis and progression, which provides novel biomarkers and therapeutic targets for HCC.

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The Significant Role of Cyclin D1 in the Synergistic Growth-inhibitory Effect of Combined Therapy of Vandetanib with 5-Fluorouracil for Gastric Cancer

Anticancer Research ◽

10.21873/anticanres.11092 ◽

2016 ◽

Vol 36 (10) ◽

pp. 5215-5226 ◽

Cited By ~ 3

Author(s):

KAZUNORI YAWATA ◽

SHINJI OSADA ◽

TOSHIYUKI TANAHASHI ◽

SATOSHI MATSUI ◽

YOSHIYUKI SASAKI ◽

...

Keyword(s):

Gastric Cancer ◽

Cyclin D1 ◽

Significant Role ◽

Combined Therapy ◽

Inhibitory Effect ◽

Growth Inhibitory Effect ◽

Growth Inhibitory

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MicroRNA-20a Suppresses Tumor Proliferation and Metastasis in Hepatocellular Carcinoma by Directly Targeting EZH1

Frontiers in Oncology ◽

10.3389/fonc.2021.737986 ◽

2021 ◽

Vol 11 ◽

Author(s):

Qianqian Zhang ◽

Xiaohong Deng ◽

Xiuxin Tang ◽

Ying You ◽

Meihua Mei ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Cell Lines ◽

Inhibitory Effect ◽

Luciferase Assay ◽

Migration And Invasion ◽

Tumor Proliferation ◽

Huh7 Cells ◽

Proliferation And Metastasis ◽

Tumor Proliferation And Metastasis

PurposeHepatocellular carcinoma (HCC), a worldwide leading cause of morbidity and mortality, is the most frequent primary liver tumor. Most HCC patients are diagnosed with advanced liver cancer, resulting in a very low 5-year survival rate. Thus, there is an urgent need for the development of targeted therapies. In this study, we aimed to investigate the effect and mechanism of the miR-20a/EZH1 axis on the proliferation and metastasis of HCC and the inhibitory effect of the EZH1/EZH2 inhibitor UNC1999 on HCC.Materials and MethodsThe expression of miR-20a in human HCC tissues and cell lines was detected using quantitative real-time PCR (qRT-PCR). The expressions of proteins were analyzed with immunohistochemistry and Western blotting. Luciferase assay was used to verify whether miR-20a targets EZH1 or EZH2. The effect of miR-20a on HCC progression was studied in vivo and in vitro. The tumor inhibitory effect of UNC1999 was confirmed in vivo. CCK8 assay, wound healing assay, cell migration and invasion assay were used to evaluate the synergistic effect of UNC1999 with sorafenib. RNA sequencing (RNA-seq) was performed to screen the differentially expressed genes in the Huh7 and SMMC7721 cell lines after UNC1999, sorafenib, and combination treatments.ResultsIn this study, miR-20a showed a lower expression in both HCC tissues and cell lines. MiR-20a inhibited the proliferation and migration of SMMC7721 and Huh7 cells. The results of the luciferase assay and Western blot analysis revealed that miR-20a directly targeted EZH1, a histone methyltransferase. We demonstrated that miR-20a negatively regulated the expression of EZH1 and inhibited the proliferation and metastasis of HCC by reducing H3K27 methylation. We found UNC1999 inhibited tumor cells proliferation and enhanced the inhibitory effect of sorafenib.ConclusionWe demonstrated that miR-20a suppresses the tumor proliferation and metastasis in HCC by directly targeting EZH1. UNC1999 can inhibit tumor proliferation in vivo and increase the sensitivity of hepatoma cell lines to sorafenib.

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The Role of HSP70, CD34 and Ki 67 Expression in Liver Cirrhosis and Hepatocellular Carcinoma, Immunohistochemical Study

Egyptian Academic Journal of Biological Sciences D Histology & Histochemistry ◽

10.21608/eajbsd.2018.85875 ◽

2018 ◽

Vol 10 (1) ◽

pp. 91-101

Author(s):

Sayed Abdel Raheem, ◽

Abdel Naby Saied ◽

Emadeldin Mater

Keyword(s):

Hepatocellular Carcinoma ◽

Liver Cirrhosis ◽

Immunohistochemical Study ◽

Ki 67

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Intra-tumoral distribution of Ki-67 and Cyclin D1 in ER+ mammary carcinoma: quantitative evaluation

African Health Sciences ◽

10.4314/ahs.v21i1.7 ◽

2021 ◽

Vol 21 (1) ◽

pp. 41-6

Author(s):

Mohammedi Latifa ◽

Djillali Doula Fatima ◽

Mesli Farida ◽

Senhadji Rachid

Keyword(s):

Breast Cancer ◽

Cyclin D1 ◽

Poor Response ◽

Labeling Index ◽

Intratumor Heterogeneity ◽

Cellular Mechanisms ◽

Ki 67 ◽

Heterogeneous Distribution ◽

Er Positive

Background: In spite of the strong evidence demonstrating the role of overexpression of Ki-67 and Cyclin D1 markers in breast carcinomas, clinical and pathological data remain to be discussed. This can be explained partly by intratumor het- erogeneity. Objectives: To define the prevalence and clinical significance of Ki-67 and Cyclin D1 overexpression in primary breast tumors ER positive, while highlighting the existence of intratumor heterogeneity in this type of cancer. Materials and methods: 51 ER positive breast cancer tumors were used to evaluate the intratumoral distribution of Ki-67 and Cyclin D1 expression. Image acquisition and visualization of the markers were performed by optical microscopy and stereology sampling method. Results: The mean Ki-67 labeling index was distributed heterogeneously in the same tumor, from 20.67±6.87 to 45.10±10.65. The coefficient of variation (COV) revealed dispersion values between 13.4% and 42.9%. Associated with positive ER status, all the tumors presented a Cyclin D1 expression with a COV varying between 19% and 28.5% and a mean labeling index fluctuating between 19.40±4.42 and 41.64±10.08 within the same patient showing important intratumor heterogeneous distribution. Conclusion: In this study, we have adopted a strictly quantitative approach to evaluate and demonstrate intratumor hetero- geneity. This establishes one of the main factors for poor response to cancer therapy. To achieve this, intratumor heteroge- neity should be usually definable and quantifiable but this domain awaits future progress and methods need to move towards a better understanding of molecular and cellular mechanisms that initiate and maintain this tumor heterogeneity. Keywords: Breast cancer; Cyclin D1; ER+; Intra-tumoral heterogeneity; Ki-67.

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microR-505/heterogeneous nuclear ribonucleoprotein M inhibits hepatocellular carcinoma cell proliferation and induces cell apoptosis through the Wnt/β-catenin signaling pathway

Biomarkers in Medicine ◽

10.2217/bmm-2019-0511 ◽

2020 ◽

Vol 14 (11) ◽

pp. 981-996

Author(s):

Xiaobin Chi ◽

Yi Jiang ◽

Yongbiao Chen ◽

Lizhi Lv ◽

Jianwei Chen ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Cell Proliferation ◽

Signaling Pathway ◽

Cell Apoptosis ◽

Heterogeneous Nuclear Ribonucleoprotein ◽

Apoptosis Assay ◽

Proliferation And Metastasis ◽

Novel Strategy ◽

Nuclear Ribonucleoprotein

Aim: This study aimed to investigate the expression of microRNA-505 (miR-505) and explore its clinical significance, biological function and mechanisms in hepatocellular carcinoma (HCC). Methods: Expression of miR-505 was measured in 128 paired HCC tissues and five cell lines by quantitative real-time polymerase chain reaction (qRT-PCR). MTT assay, Transwell migration, invasion assays and apoptosis assay were performed to explore the functional role of miR-505. The target gene of miR-505 was assessed using the bioinformatics assay and the related signaling pathway was confirmed using western blot. Results: Expression of miR-505 in HCC serum and tissues were downregulated. The overexpression of miR-505 in HCC cells inhibited cell proliferation and metastasis, as well as enhanced cell apoptosis by directly downregulating heterogeneous nuclear ribonucleoprotein M ( HNRNPM). The activity of the Wnt/β-catenin signaling pathway was suppressed by the overexpression of miR-505 but was promoted by the upregulation of HNRNPM. Conclusion: The results suggest that the regulation of miR-505/ HNRNPM may be a novel strategy to improve the targeted therapy of HCC.

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