Potential Chemopreventive role of Boldine against Hepatocellular Carcinoma via modulation of Cell Cycle Proteins in Rat Model

2021 ◽  
Vol 21 ◽  
Author(s):  
Nirmala Subramaniam ◽  
Pugazhendhi Kannan ◽  
Jagan Sundaram ◽  
Ashok Mari ◽  
Sathesh K. Velli ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Cell Cycle ◽  
Albino Rats ◽  
Oxygen Species ◽  
Cyclin E1 ◽  
Chemopreventive Agent ◽  
Regulated Expression ◽  
Proliferative Markers ◽  
Wistar Albino Rats ◽  
Phase I And Ii

Background: To evaluate the chemopreventive potential of boldine against diethylnitrosamine (DEN) induced hepatocellular carcinoma (HCC) in wistar albino rats. Objective: Boldine is an alkaloid isolated from Peumus boldus. The primary active constituents of boldine exhibited several potential medicinal properties. The present study was evaluated to explore the chemopreventive agent of boldine on anti-proliferative efficacy against diethylnitrosamine (DEN) induced hepatocellular carcinoma (HCC) in wistar albino rats. Methods: The effect of boldine on cellular proliferative markers, i.e., PCNA and Ki67 on hepatocellular carcinoma rats was determined by immuno expression study. Liver marker enzymes, tumor biomarker, oxidative stress markers, anti-oxidant status and xenobiotic phase I and II enzymes in HCC rats were analyzed. Moreover, cell cycle proteins, i.e., p21Cip1/Kip1, p27 Cip1/Kip1, Cyclin D1, CDK 4, Cyclin E1, and CDK 2 were investigated using immuno expression analysis. Results: Treatment of boldine protected the liver against reactive oxygen species such as hydrogen peroxide, superoxide, protein carbonyl and lipid peroxide during hepatocarcinogenesis by boosted antioxidants-superoxide dismutase (SOD), catalase (CAT). Boldine caused substantial enhanced detoxification process by moderating phase I and II xenobiotic metabolizing enzymes. In addition, the study was found that boldine significantly inhibited the cellular proliferative markers like PCNA and Ki67 and regulated the specific cell cycle associated proteins by up-regulated expression of p21Cip1/Kip1 and p27 Cip1/Kip1 and down-regulated expression of Cyclin D1, CDK 4, Cyclin E1, and CDK 2. Conclusion: Our data manifests the anti-proliferative effect of boldine, which negatively modulates cellular proliferation and regulates cell cycle by protecting the cell from reactive oxygen species (ROS), suggesting that boldine establish it as a chemopreventive agent in diethylnitrosamine-induced hepatocarcinogenesis in rats.

scholarly journals Cyclin E1 and cyclin-dependent kinase 2 are critical for initiation, but not for progression of hepatocellular carcinoma

2018 ◽  
Vol 115 (37) ◽  
pp. 9282-9287 ◽  
Author(s):  
Roland Sonntag ◽  
Nives Giebeler ◽  
Yulia A. Nevzorova ◽  
Jörg-Martin Bangen ◽  
Dirk Fahrenkamp ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Cell Cycle ◽  
Gene Signature ◽  
Cyclin Dependent Kinase ◽  
Regulatory Subunits ◽  
Cyclin E1 ◽  
Quiescent Cells ◽  
Similar Expression Profile ◽  
Regulation Of Cell Cycle

E-type cyclins E1 (CcnE1) and E2 (CcnE2) are regulatory subunits of cyclin-dependent kinase 2 (Cdk2) and thought to control the transition of quiescent cells into the cell cycle. Initial findings indicated that CcnE1 and CcnE2 have largely overlapping functions for cancer development in several tumor entities including hepatocellular carcinoma (HCC). In the present study, we dissected the differential contributions of CcnE1, CcnE2, and Cdk2 for initiation and progression of HCC in mice and patients. To this end, we tested the HCC susceptibility in mice with constitutive deficiency for CcnE1 or CcnE2 as well as in mice lacking Cdk2 in hepatocytes. Genetic inactivation of CcnE1 largely prevented development of liver cancer in mice in two established HCC models, while ablation of CcnE2 had no effect on hepatocarcinogenesis. Importantly, CcnE1-driven HCC initiation was dependent on Cdk2. However, isolated primary hepatoma cells typically acquired independence on CcnE1 and Cdk2 with increasing progression in vitro, which was associated with a gene signature involving secondary induction of CcnE2 and up-regulation of cell cycle and DNA repair pathways. Importantly, a similar expression profile was also found in HCC patients with elevated CcnE2 expression and poor survival. In general, overall survival in HCC patients was synergistically affected by expression of CcnE1 and CcnE2, but not through Cdk2. Our study suggests that HCC initiation specifically depends on CcnE1 and Cdk2, while HCC progression requires expression of any E-cyclin, but no Cdk2.

scholarly journals Regulation of cell cycle of hepatocellular carcinoma by NF90 through modulation of cyclin E1 mRNA stability

Oncogene ◽  
2014 ◽  
Vol 34 (34) ◽  
pp. 4460-4470 ◽  
Author(s):  
W Jiang ◽  
H Huang ◽  
L Ding ◽  
P Zhu ◽  
H Saiyin ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Cell Cycle ◽  
Mrna Stability ◽  
Cyclin E1 ◽  
Regulation Of Cell Cycle

scholarly journals Tamarix articulata Inhibits Cell Proliferation, Promotes Cell Death Mechanisms and Triggers G0/G1 Cell Cycle Arrest in Hepatocellular Carcinoma Cells

2021 ◽  
Vol 59 (2) ◽  
Author(s):  
Abdullah Mohammad Alnuqaydan ◽  
Bilal Rah
Keyword(s):  
Hepatocellular Carcinoma ◽  
Reactive Oxygen Species ◽  
Cell Cycle ◽  
Cell Death ◽  
Methanolic Extract ◽  
Reactive Oxygen ◽  
Carcinoma Cells ◽  
Oxygen Species ◽  
Hepatocellular Carcinoma Cells ◽  
Acridine Orange Staining

Research background. From ancient times plants have been used for medicinal purposes against various ailments. In the modern era, plants are a major source of drugs and are an appealing drug candidate for the anticancer therapeutics against various molecular targets. Here we tested Tamarix articulata methanolic extract of dry leaves for anticancer activity against a panel of hepatocellular carcinoma cells. Experimental approach. Cell viability of hepatocellular carcinoma cells was determined by MTT assay after dose-dependent treatment with extract of Tamarix articulata. Phase-contrast microscopy and DAPI staining were performed to analyze cellular and nuclear morphology. Immunoblotting was performed to determine the expression of proteins associated with autophagy, apoptosis, and cell cycle. However, flow cytometry was used for the quantification of apoptotic cells and the analysis of cells in different phases of the cell cycle after treatment with varying doses of Tamarix articulata. Additionally, acridine orange staining and DCFHDA dye were used to analyze the quantification of autophagosomes and reactive oxygen species. Results and conclusion. Our results demonstrate that Tamarix articulata methanolic extract exhibits promising antiproliferative activity with IC50 values (271±4.38), (298±7.08) and (336±6.11) µg/mL against HepG2, Huh7D12, and Hep3B cell lines, respectively. Mechanistically, we found Tamarix articulata methanolic extract induces cell death by activating apoptosis and autophagy pathways. First, Tamarix articulata methanolic extract promotes autophagy which was confirmed by acridine orange staining. The immunoblotting analysis further confirms that Tamarix articulata methanolic extract consistently induces the conversion of, LC3I to LC3II form with a gradual decrease in expression of autophagy substrate protein p62 at higher doses. Second, Tamarix articulata methanolic extract promotes reactive oxygen species production in hepatocellular carcinoma cells and activates reactive oxygen species-mediated apoptosis. Flow cytometry and immunoblotting analysis showed that Tamarix articulata methanolic extract induces dose-dependent apoptosis and activates proapoptotic proteins caspase-3 and PARP1. Additionally, Tamarix articulata methanolic extract triggers the arrest of the G0/G1 phase of the cell cycle and upregulates the protein expression of p27/Kip, and p21/Cip, with a decrease in cyclin-D1 expression in hepatocellular carcinoma cells. Novelty and scientific contribution. The current study demonstrates that Tamarix articulata methanolic extract exhibits promising anticancer potential to kill tumor cells by programmed-cell-death type I and II mechanisms and could be explored for potential drug candidate molecules to curtail cancer in the future.

Lappaconitine Sulfate Inhibits Proliferation and Induces Apoptosis in Human Hepatocellular Carcinoma HepG2 Cells through the Reactive Oxygen Species-Dependent Mitochondrial Pathway

Pharmacology ◽  
2020 ◽  
Vol 105 (11-12) ◽  
pp. 705-714
Author(s):  
Xuemei Zhang ◽  
Junyi Ma ◽  
Na Song ◽  
Yongyue Guo ◽  
Ling Hui ◽  
...  
Keyword(s):  
Hepatocellular Carcinoma ◽  
Reactive Oxygen Species ◽  
Cell Cycle ◽  
Cell Cycle Arrest ◽  
Hepg2 Cells ◽  
Reactive Oxygen ◽  
Oxygen Species ◽  
Antitumor Activities ◽  
Cycle Arrest

<b><i>Background:</i></b> Hepatocellular carcinoma (HCC) is the third leading cause of tumor-related deaths in the word. Lappaconitine (LA), a diterpenoid alkaloid, exerts antitumor activities. However, the effects and mechanisms of LA sulfate (LS) on HCC remain unclear. This study evaluated the activities and explored the underlying mechanisms of LS in HCC cell line HepG2 cells. <b><i>Materials and Methods:</i></b> The cell viability and proliferation were evaluated using the Cell Counting Kit-8 (CCK-8) and 5-ethynyl-2′-deoxyuridine (EdU) assay, respectively. The cell cycle distribution was detected by propidium iodide (PI) staining assay. The apoptosis was detected by Annexin ­V-fluorescein isothiocyanate (FITC)/PI double staining assay. The cell cycle arrest and apoptosis-related proteins were estimated by western blot analysis. The mitochondrial membrane potential (MMP) was ­determined through the 5, 5′, 6, 6′-tetrachloro-1, 1′, 3, 3′-tetraethylbenzimi-dazolyl carbocyanine iodide (JC-1) staining assay. The reactive oxygen species (ROS) was monitored by 20–70-dichlorofluorescein diacetate (DCFH-DA) staining assay. In vivo antitumor activities were investigated by HepG2 xenograft model. <b><i>Results:</i></b> Our results showed that LS significantly ­inhibited the viability and proliferation of HepG2 cells. LS triggered G0/G1 cell cycle arrest, apoptosis and caspase activation. Furthermore, LS induced MMP loss and ROS accumulation. Additionally, LS suppressed the phosphatidylinositol-3-kinase (PI3K)/protein kinase B (AKT)/glycogen synthase kinase 3β (GSK3β) signaling pathway. An in vivo assay showed that LS exhibited a pronounced antitumor effect in nude mice bearing HepG2 xenografts. <b><i>Conclusions:</i></b> Our results demonstrated that LS is a promising therapeutic agent for HCC directed ­toward the proliferation inhibition and the induction of apoptosis.

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