Interventional gene targeting of cell cycle regulators identifies Cyclin E1 as a suitable target for attenuating hepatocellular carcinoma progression

Hepatocellular carcinoma (HCC) is the second and sixth leading cause of cancer death in men and woman in 185 countries statistics, respectively. n-Butylidenephthalide (BP) has shown anti-HCC activity, but it also has an unstable structure that decreases its potential antitumor activity. The aim of this study was to investigate the cell uptake, activity protection, and antitumor mechanism of BP encapsulated in the novel liposome LPPC in HCC cells. BP/LPPC exhibited higher cell uptake and cytotoxicity than BP alone, and combined with clinical drug etoposide (VP-16), BP/LPPC showed a synergistic effect against HCC cells. Additionally, BP/LPPC increased cell cycle regulators (p53, p-p53, and p21) and decreased cell cycle-related proteins (Rb, p-Rb, CDK4, and cyclin D1), leading to cell cycle arrest at the G0/G1 phase in HCC cells. BP/LPPC induced cell apoptosis through activation of both the extrinsic (Fas-L and Caspase-8) and intrinsic (Bax and Caspase-9) apoptosis pathways and activated the caspase cascade to trigger HCC cell death. In conclusion, the LPPC complex improved the antitumor activity of BP in terms of cytotoxicity, cell cycle regulation and cell apoptosis, and BP/LPPC synergistically inhibited cell growth during combination treatment with VP-16 in HCC cells. Therefore, BP/LPPC is potentially a good candidate for clinical drug development or for use as an adjuvant for clinical drugs as a combination therapy for hepatocellular carcinoma.

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

Proceedings of the National Academy of Sciences ◽

10.1073/pnas.1807155115 ◽

2018 ◽

Vol 115 (37) ◽

pp. 9282-9287 ◽

Cited By ~ 13

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.

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Signficance of cell cycle regulators in human hepatocellular carcinoma and gene expression induced by cisplatin in hepatoma cell lines

10.5353/th_b3124224 ◽

2000 ◽

Author(s):

Lanfang Qin

Keyword(s):

Gene Expression ◽

Hepatocellular Carcinoma ◽

Cell Cycle ◽

Cell Lines ◽

Hepatoma Cell ◽

Human Hepatocellular Carcinoma ◽

Cell Cycle Regulators ◽

Hepatoma Cell Lines

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

Anti-Cancer Agents in Medicinal Chemistry ◽

10.2174/1871520621666210203102854 ◽

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.

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SKP2 and CKS1 Promote Degradation of Cell Cycle Regulators and Are Associated With Hepatocellular Carcinoma Prognosis

Gastroenterology ◽

10.1053/j.gastro.2009.08.005 ◽

2009 ◽

Vol 137 (5) ◽

pp. 1816-1826.e10 ◽

Cited By ~ 70

Author(s):

Diego F. Calvisi ◽

Sara Ladu ◽

Federico Pinna ◽

Maddalena Frau ◽

Maria L. Tomasi ◽

...

Keyword(s):

Hepatocellular Carcinoma ◽

Cell Cycle ◽

Cell Cycle Regulators

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DDX3 Regulates Cell Growth through Translational Control of Cyclin E1

Molecular and Cellular Biology ◽

10.1128/mcb.00560-10 ◽

2010 ◽

Vol 30 (22) ◽

pp. 5444-5453 ◽

Cited By ~ 96

Author(s):

Ming-Chih Lai ◽

Wen-Cheng Chang ◽

Sheau-Yann Shieh ◽

Woan-Yuh Tarn

Keyword(s):

Cell Cycle ◽

Cell Growth ◽

Translational Control ◽

Cell Cycle Control ◽

Temperature Sensitive ◽

Alternative Mechanism ◽

Cell Cycle Regulators ◽

Rna Helicases ◽

Cyclin E1 ◽

Mrna Targets

ABSTRACT DDX3 belongs to the DEAD box family of RNA helicases, but the details of its biological function remain largely unclear. Here we show that knockdown of DDX3 expression impedes G1/S-phase transition of the cell cycle. To know how DDX3 may act in cell cycle control, we screened for cellular mRNA targets of DDX3. Many of the identified DDX3 targets encoded cell cycle regulators, including G1/S-specific cyclin E1. DDX3 depletion specifically downregulates translation of cyclin E1 mRNA. Moreover, our data suggest that DDX3 participates in translation initiation of targeted mRNAs as well as in cell growth control via its RNA helicase activity. Consistent with these findings, we show that in the temperature-sensitive DDX3 mutant hamster cell line tsET24, cyclin E1 expression is downregulated at a nonpermissive temperature that inactivates mutant DDX3. Taken together, our results indicate that DDX3 is critical for translation of cyclin E1 mRNA, which provides an alternative mechanism for regulating cyclin E1 expression during the cell cycle.

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