International Journal of Hematology and Oncology
2023, Vol 33, Num 4 Page(s): 146-152
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Investigation of the Association Between TET2 Expression and Response to CAPE
Burcu YUCEL1, Muhammed Ali FURAL2, Bircan SONMEZ3, Ozlen BEKTAS4, Mehmet SONMEZ4
1Istanbul Medeniyet University, Faculty of Medicine, Department of Medical Biology, Istanbul, TURKEY
2Karadeniz Technical University, Faculty of Medicine, Department of Internal Medicine, Trabzon, TURKEY
3Karadeniz Technical University, Faculty of Medicine, Department of Nuclear Medicine, Trabzon, TURKEY
4Karadeniz Technical University, Faculty of Medicine, Department of Internal Medicine, Division of Hematology, Trabzon, TURKEY
Keywords: Leukemia, TET2 expression, shRNA-mediated gene silencing, Anti-cancer
Acute myeloid leukemia (AML) is a clonal, neoplastic disease characterized by abnormal proliferation of myeloid progenitor cells. Genetic and epigenetic changes in AML patients impair proliferation and differentiation of myeloid progenitor cells. TET2 functions in DNA demethylation and mutations are frequently observed in patients with AML. Caffeic acid phenethyl ester (CAPE) is an active component of propolis, a resinous substance collected by honey bees from various plant sources. CAPE’s antioxidant, anti-inflammatory, antiviral, immunostimulant and anticancer effects have been shown in various studies. In this study, we aimed to investigate CAPE’s effect on TET2 mRNA expression. In K562 cell line, TET2 mRNA expression upon CAPE treatment was controlled with qRT-PCR. To decrease TET2 mRNA expression commercially available TET2 mRNA targeting shRNAs and a control shRNA were used. Transfection grade plasmids were then used to transfect K562 cells and antibiotic selection was applied for stable transfection. CAPE activity on cell viability in TET2 downregulated cells was assessed with Wst-1 assay. 5mM CAPE treatment increased TET2 mRNA expression fold change up to 2.5 times (p< 0.05). Two different shRNAs downregulated TET2 mRNA expression almost 50% compare to control plasmid (p< 0.05). TET2 downregulated cells were found more resistant to CAPE treatment (shRNA1; p< 0.0001 and shRNA2; p< 0.05). Our findings suggest that CAPE acts on cells by changing TET2 mRNA expression in K562 leukemia cells. In addition, TET2 downregulated cells were found more resistant to CAPE suggesting that TET2 expression may play a role in response to conventional treatments.
Burcu YUCEL1, Muhammed Ali FURAL2, Bircan SONMEZ3, Ozlen BEKTAS4, Mehmet SONMEZ4
1Istanbul Medeniyet University, Faculty of Medicine, Department of Medical Biology, Istanbul, TURKEY
2Karadeniz Technical University, Faculty of Medicine, Department of Internal Medicine, Trabzon, TURKEY
3Karadeniz Technical University, Faculty of Medicine, Department of Nuclear Medicine, Trabzon, TURKEY
4Karadeniz Technical University, Faculty of Medicine, Department of Internal Medicine, Division of Hematology, Trabzon, TURKEY
Keywords: Leukemia, TET2 expression, shRNA-mediated gene silencing, Anti-cancer
Acute myeloid leukemia (AML) is a clonal, neoplastic disease characterized by abnormal proliferation of myeloid progenitor cells. Genetic and epigenetic changes in AML patients impair proliferation and differentiation of myeloid progenitor cells. TET2 functions in DNA demethylation and mutations are frequently observed in patients with AML. Caffeic acid phenethyl ester (CAPE) is an active component of propolis, a resinous substance collected by honey bees from various plant sources. CAPE’s antioxidant, anti-inflammatory, antiviral, immunostimulant and anticancer effects have been shown in various studies. In this study, we aimed to investigate CAPE’s effect on TET2 mRNA expression. In K562 cell line, TET2 mRNA expression upon CAPE treatment was controlled with qRT-PCR. To decrease TET2 mRNA expression commercially available TET2 mRNA targeting shRNAs and a control shRNA were used. Transfection grade plasmids were then used to transfect K562 cells and antibiotic selection was applied for stable transfection. CAPE activity on cell viability in TET2 downregulated cells was assessed with Wst-1 assay. 5mM CAPE treatment increased TET2 mRNA expression fold change up to 2.5 times (p< 0.05). Two different shRNAs downregulated TET2 mRNA expression almost 50% compare to control plasmid (p< 0.05). TET2 downregulated cells were found more resistant to CAPE treatment (shRNA1; p< 0.0001 and shRNA2; p< 0.05). Our findings suggest that CAPE acts on cells by changing TET2 mRNA expression in K562 leukemia cells. In addition, TET2 downregulated cells were found more resistant to CAPE suggesting that TET2 expression may play a role in response to conventional treatments.
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