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Curcumin and 6-Shogaol Increase Hemoglobin F Levels by Inhibiting Expression of STAT3 mRNA Gene in K562 Line Cell


Affiliations
1 Department of Biochemistry, Faculty of Medicine, University of Jenderal Soedirman, Purwokerto, Indonesia
2 Department of Biochemistry, Faculty of Medicine, Public Health and Nursing, University of Gadjah Mada, Jogjakarta, Indonesia
3 Cancer Chemoprevention Research Center, Faculty of Pharmacy, University of Gadjah Mada, Jogjakarta, Indonesia
4 Department of Pharmacology, Faculty of Medicine, Public Health and Nursing, University of Gadjah Mada, Jogjakarta, Indonesia
     

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One of the approaches for beta-thalassemia therapy is the induction of the Haemoglobin F (Hb F). Curcumin and 6-Shogaol are empirically known to induce HbF, but the signalling cascade has not been widely explained. This study aims to uncover the potential of Curcumin and 6-shogaol in inhibiting the expression of STAT3 mRNA gene.This study uses the K562 erythroleukemic line cell model with an experimental design post-test only with a control group. There are 5 groups, each group has 3 replications, named the control group without treatment, the positive control group with Hydroxyurea treatment (75 μM), the combination treatment group of Curcumin (2 μM) and 6-Shogaol (10 μM), the single curcumin (2 μM) treatment group and a single 6-Shogaol (10 μM) treatment group. Test samples were taken in 72-h and 96-h time series, then RNA extraction from the cell line was continued by cDNA synthesis. The expression of STAT3 mRNA gene was measured using the qRT-PCR technique; then, the Hb F level was measured by the ELISA method. Statistical analysis using ANOVA test with significance level p <0.05. In the 72-h time series, there was a significant decrease in STAT3 Gena mRNA expression (p <0.05). The lowest single curcumin group (p <0.01) followed by a single 6-Shogaol group (p <0.05) compared to the untreated control group, while the positive control group with hydroxyurea treatment and the Curcumin and 6-shogaol combination treatment groups are not significant. Hb F levels, there was an increase in 96-h time series (p <0.05) respectively from highest to lowest in the curcumin group (p <0.05), 6-shogaol (p> 0.05) compared to the control group, but in the positive control group (p> 0.05) and the combination group (p> 0.05) it is lower than the control group without any treatment. Curcumin and 6-shogaol increase Hb F levels through inhibition expression of STAT3 mRNA Gene on K562 cells. The results of this study could be the basis for further research in vivo to reveal the signalling pathway in Hb Finduction therapy.

Keywords

Curcumin, 6-shogaol, STAT3 Gena mRNA, Hemoglobin F, K562 cells
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  • Curcumin and 6-Shogaol Increase Hemoglobin F Levels by Inhibiting Expression of STAT3 mRNA Gene in K562 Line Cell

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Authors

Joko Setyono
Department of Biochemistry, Faculty of Medicine, University of Jenderal Soedirman, Purwokerto, Indonesia
Ahmad Hamim Sadewa
Department of Biochemistry, Faculty of Medicine, Public Health and Nursing, University of Gadjah Mada, Jogjakarta, Indonesia
Edy Meiyanto
Cancer Chemoprevention Research Center, Faculty of Pharmacy, University of Gadjah Mada, Jogjakarta, Indonesia
Mustofa Mustofa
Department of Pharmacology, Faculty of Medicine, Public Health and Nursing, University of Gadjah Mada, Jogjakarta, Indonesia

Abstract


One of the approaches for beta-thalassemia therapy is the induction of the Haemoglobin F (Hb F). Curcumin and 6-Shogaol are empirically known to induce HbF, but the signalling cascade has not been widely explained. This study aims to uncover the potential of Curcumin and 6-shogaol in inhibiting the expression of STAT3 mRNA gene.This study uses the K562 erythroleukemic line cell model with an experimental design post-test only with a control group. There are 5 groups, each group has 3 replications, named the control group without treatment, the positive control group with Hydroxyurea treatment (75 μM), the combination treatment group of Curcumin (2 μM) and 6-Shogaol (10 μM), the single curcumin (2 μM) treatment group and a single 6-Shogaol (10 μM) treatment group. Test samples were taken in 72-h and 96-h time series, then RNA extraction from the cell line was continued by cDNA synthesis. The expression of STAT3 mRNA gene was measured using the qRT-PCR technique; then, the Hb F level was measured by the ELISA method. Statistical analysis using ANOVA test with significance level p <0.05. In the 72-h time series, there was a significant decrease in STAT3 Gena mRNA expression (p <0.05). The lowest single curcumin group (p <0.01) followed by a single 6-Shogaol group (p <0.05) compared to the untreated control group, while the positive control group with hydroxyurea treatment and the Curcumin and 6-shogaol combination treatment groups are not significant. Hb F levels, there was an increase in 96-h time series (p <0.05) respectively from highest to lowest in the curcumin group (p <0.05), 6-shogaol (p> 0.05) compared to the control group, but in the positive control group (p> 0.05) and the combination group (p> 0.05) it is lower than the control group without any treatment. Curcumin and 6-shogaol increase Hb F levels through inhibition expression of STAT3 mRNA Gene on K562 cells. The results of this study could be the basis for further research in vivo to reveal the signalling pathway in Hb Finduction therapy.

Keywords


Curcumin, 6-shogaol, STAT3 Gena mRNA, Hemoglobin F, K562 cells



DOI: https://doi.org/10.37506/v11%2Fi2%2F2020%2Fijphrd%2F194994