The Synergistic Antitumor Effect of Decitabine and Vorinostat Combination on HepG2 Human Hepatocellular Carcinoma Cell Line via Epigenetic Modulation of Autophagy–Apoptosis Molecular Crosstalk
Hepatocellular carcinoma (HCC) is a worldwide health issue. Epigenetic alterations play a crucial role in HCC tumorigenesis. Using epigenetic modulators for HCC treatment confers a promising therapeutic effect. The aim of this study was to explore the effect of a decitabine (DAC) and vorinostat (VOR...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2023-07-01
|
| Series: | Current Issues in Molecular Biology |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1467-3045/45/7/375 |
| _version_ | 1827733457989533696 |
|---|---|
| author | Basant M. Salama Maged W. Helmy Hosny Fouad Marium M. Shamaa Maha E. Houssen |
| author_facet | Basant M. Salama Maged W. Helmy Hosny Fouad Marium M. Shamaa Maha E. Houssen |
| author_sort | Basant M. Salama |
| collection | DOAJ |
| description | Hepatocellular carcinoma (HCC) is a worldwide health issue. Epigenetic alterations play a crucial role in HCC tumorigenesis. Using epigenetic modulators for HCC treatment confers a promising therapeutic effect. The aim of this study was to explore the effect of a decitabine (DAC) and vorinostat (VOR) combination on the crosstalk between apoptosis and autophagy in the HCC HepG2 cell line at 24 h and 72 h. Median inhibitory concentrations (IC<sub>50</sub>s) of VOR and DAC were assessed in the HepG2 cell line. The activity of caspase-3 was evaluated colorimetrically, and Cyclin D1(CCND1), Bcl-2, ATG5, ATG7, and P62 levels were assessed using ELISA at different time intervals (24 h and 72 h), while <i>LC3IIB</i> and <i>Beclin-1</i>gene expression were measured by using qRT-PCR. The synergistic effect of VOR and DAC was confirmed due to the observed combination indices (CIs) and dose reduction indices (DRIs). The combined treatment with both drugs inhibited the proliferation marker (CCND1), and enhanced apoptosis compared with each drug alone at 24 h and 72 h <i>(</i>via active caspase-3 upregulation and Bcl-2 downregulation). Moreover, the combination induced autophagy as an early event via upregulation of <i>Beclin-1</i>, <i>LC3IIB</i>, ATG5, and ATG7 gene expression. The initial induction of autophagy started to decrease after 72 h due to <i>Beclin-1</i> downregulation, and there was decreased expression of <i>LC3IIB</i> compared with the value at 24 h. Herein, epigenetic modulation via the VOR/DAC combination showed an antitumor effect through the coordination of an autophagy–apoptosis crosstalk and promotion of autophagy-induced apoptosis, which ultimately led to the cellular death of HCC cancer cells. |
| first_indexed | 2024-03-11T01:10:39Z |
| format | Article |
| id | doaj.art-5ca409d2b6d54e3382d317602c37edc7 |
| institution | Directory Open Access Journal |
| issn | 1467-3037 1467-3045 |
| language | English |
| last_indexed | 2024-03-11T01:10:39Z |
| publishDate | 2023-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Current Issues in Molecular Biology |
| spelling | doaj.art-5ca409d2b6d54e3382d317602c37edc72023-11-18T18:51:22ZengMDPI AGCurrent Issues in Molecular Biology1467-30371467-30452023-07-014575935594910.3390/cimb45070375The Synergistic Antitumor Effect of Decitabine and Vorinostat Combination on HepG2 Human Hepatocellular Carcinoma Cell Line via Epigenetic Modulation of Autophagy–Apoptosis Molecular CrosstalkBasant M. Salama0Maged W. Helmy1Hosny Fouad2Marium M. Shamaa3Maha E. Houssen4Department of Biochemistry, Faculty of Pharmacy, Damanhour University, Damanhour 22511, EgyptPharmacology and Toxicology Department, Faculty of Pharmacy, Damanhour University, Damanhour 22511, EgyptPharmacology Department, Faculty of Pharmacy, Alexandria University, Alexandria 21521, EgyptDepartment of Biochemistry, Clinical and Biological Science Division, College of Pharmacy, Arab Academy for Science, Technology and Maritime Transport, Alexandria 1029, EgyptDepartment of Biochemistry, Faculty of Pharmacy, Damanhour University, Damanhour 22511, EgyptHepatocellular carcinoma (HCC) is a worldwide health issue. Epigenetic alterations play a crucial role in HCC tumorigenesis. Using epigenetic modulators for HCC treatment confers a promising therapeutic effect. The aim of this study was to explore the effect of a decitabine (DAC) and vorinostat (VOR) combination on the crosstalk between apoptosis and autophagy in the HCC HepG2 cell line at 24 h and 72 h. Median inhibitory concentrations (IC<sub>50</sub>s) of VOR and DAC were assessed in the HepG2 cell line. The activity of caspase-3 was evaluated colorimetrically, and Cyclin D1(CCND1), Bcl-2, ATG5, ATG7, and P62 levels were assessed using ELISA at different time intervals (24 h and 72 h), while <i>LC3IIB</i> and <i>Beclin-1</i>gene expression were measured by using qRT-PCR. The synergistic effect of VOR and DAC was confirmed due to the observed combination indices (CIs) and dose reduction indices (DRIs). The combined treatment with both drugs inhibited the proliferation marker (CCND1), and enhanced apoptosis compared with each drug alone at 24 h and 72 h <i>(</i>via active caspase-3 upregulation and Bcl-2 downregulation). Moreover, the combination induced autophagy as an early event via upregulation of <i>Beclin-1</i>, <i>LC3IIB</i>, ATG5, and ATG7 gene expression. The initial induction of autophagy started to decrease after 72 h due to <i>Beclin-1</i> downregulation, and there was decreased expression of <i>LC3IIB</i> compared with the value at 24 h. Herein, epigenetic modulation via the VOR/DAC combination showed an antitumor effect through the coordination of an autophagy–apoptosis crosstalk and promotion of autophagy-induced apoptosis, which ultimately led to the cellular death of HCC cancer cells.https://www.mdpi.com/1467-3045/45/7/375DNA methyltransferase inhibitorsdecitabinehistone deacetylase inhibitorsvorinostatapoptosisautophagy |
| spellingShingle | Basant M. Salama Maged W. Helmy Hosny Fouad Marium M. Shamaa Maha E. Houssen The Synergistic Antitumor Effect of Decitabine and Vorinostat Combination on HepG2 Human Hepatocellular Carcinoma Cell Line via Epigenetic Modulation of Autophagy–Apoptosis Molecular Crosstalk Current Issues in Molecular Biology DNA methyltransferase inhibitors decitabine histone deacetylase inhibitors vorinostat apoptosis autophagy |
| title | The Synergistic Antitumor Effect of Decitabine and Vorinostat Combination on HepG2 Human Hepatocellular Carcinoma Cell Line via Epigenetic Modulation of Autophagy–Apoptosis Molecular Crosstalk |
| title_full | The Synergistic Antitumor Effect of Decitabine and Vorinostat Combination on HepG2 Human Hepatocellular Carcinoma Cell Line via Epigenetic Modulation of Autophagy–Apoptosis Molecular Crosstalk |
| title_fullStr | The Synergistic Antitumor Effect of Decitabine and Vorinostat Combination on HepG2 Human Hepatocellular Carcinoma Cell Line via Epigenetic Modulation of Autophagy–Apoptosis Molecular Crosstalk |
| title_full_unstemmed | The Synergistic Antitumor Effect of Decitabine and Vorinostat Combination on HepG2 Human Hepatocellular Carcinoma Cell Line via Epigenetic Modulation of Autophagy–Apoptosis Molecular Crosstalk |
| title_short | The Synergistic Antitumor Effect of Decitabine and Vorinostat Combination on HepG2 Human Hepatocellular Carcinoma Cell Line via Epigenetic Modulation of Autophagy–Apoptosis Molecular Crosstalk |
| title_sort | synergistic antitumor effect of decitabine and vorinostat combination on hepg2 human hepatocellular carcinoma cell line via epigenetic modulation of autophagy apoptosis molecular crosstalk |
| topic | DNA methyltransferase inhibitors decitabine histone deacetylase inhibitors vorinostat apoptosis autophagy |
| url | https://www.mdpi.com/1467-3045/45/7/375 |
| work_keys_str_mv | AT basantmsalama thesynergisticantitumoreffectofdecitabineandvorinostatcombinationonhepg2humanhepatocellularcarcinomacelllineviaepigeneticmodulationofautophagyapoptosismolecularcrosstalk AT magedwhelmy thesynergisticantitumoreffectofdecitabineandvorinostatcombinationonhepg2humanhepatocellularcarcinomacelllineviaepigeneticmodulationofautophagyapoptosismolecularcrosstalk AT hosnyfouad thesynergisticantitumoreffectofdecitabineandvorinostatcombinationonhepg2humanhepatocellularcarcinomacelllineviaepigeneticmodulationofautophagyapoptosismolecularcrosstalk AT mariummshamaa thesynergisticantitumoreffectofdecitabineandvorinostatcombinationonhepg2humanhepatocellularcarcinomacelllineviaepigeneticmodulationofautophagyapoptosismolecularcrosstalk AT mahaehoussen thesynergisticantitumoreffectofdecitabineandvorinostatcombinationonhepg2humanhepatocellularcarcinomacelllineviaepigeneticmodulationofautophagyapoptosismolecularcrosstalk AT basantmsalama synergisticantitumoreffectofdecitabineandvorinostatcombinationonhepg2humanhepatocellularcarcinomacelllineviaepigeneticmodulationofautophagyapoptosismolecularcrosstalk AT magedwhelmy synergisticantitumoreffectofdecitabineandvorinostatcombinationonhepg2humanhepatocellularcarcinomacelllineviaepigeneticmodulationofautophagyapoptosismolecularcrosstalk AT hosnyfouad synergisticantitumoreffectofdecitabineandvorinostatcombinationonhepg2humanhepatocellularcarcinomacelllineviaepigeneticmodulationofautophagyapoptosismolecularcrosstalk AT mariummshamaa synergisticantitumoreffectofdecitabineandvorinostatcombinationonhepg2humanhepatocellularcarcinomacelllineviaepigeneticmodulationofautophagyapoptosismolecularcrosstalk AT mahaehoussen synergisticantitumoreffectofdecitabineandvorinostatcombinationonhepg2humanhepatocellularcarcinomacelllineviaepigeneticmodulationofautophagyapoptosismolecularcrosstalk |