Curcumin Analog DK1 Induces Apoptosis in Human Osteosarcoma Cells In Vitro through Mitochondria-Dependent Signaling Pathway

Abstract

Osteosarcoma is one of the primary malignant bone tumors that confer low survival rates for patients even with intensive regime treatments. Therefore, discovery of novel anti-osteosarcoma drugs derived from natural products that are not harmful to the normal cells remains crucial. Curcumin is one of the natural substances that have been extensively studied due to its anti-cancer properties and is pharmacologically safe considering its ubiquitous consumption for centuries. However, curcumin suffers from a poor circulating bioavailability, which has led to the development of a chemically synthesized curcuminoid analog, namely (Z)-3-hydroxy-1-(2-hydroxyphenyl)-3-phenylprop-2-en-1-one (DK1). In this study, the cytotoxic effects of the curcumin analog DK1 was investigated in both U-2OS and MG-63 osteosarcoma cell lines using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay and cell death was microscopically examined via acridine orange/propidium iodide (AO/PI) double staining. Flow cytometer analysis including Annexin V/Fluorescein isothiocyanate (FITC), cell cycle analysis and JC-1 were adapted to determine the mode of cell death. Subsequently in order to determine the mechanism of cell death, quantitative polymerase chain reaction (qPCR) and proteome profiling was carried out to measure the expression of several apoptotic-related genes and proteins. Results indicated that DK1 induced U-2 OS and MG-63 morphological changes and substantially reduced cell numbers through induction of apoptosis. Several apoptotic genes and proteins were steadily expressed after treatment with DK1; including caspase 3, caspase 9, and BAX, which indicated that apoptosis occurred through a mitochondria-dependent signaling pathway. In conclusion, DK1 could be considered as a potential candidate for an anti-osteosarcoma drug in the near future, contingent upon its ability to induce apoptosis in osteosarcoma cell lines.

DK1 Induces Apoptosis via Mitochondria-Dependent Signaling Pathway in Human Colon Carcinoma Cell Lines In Vitro

“…The externalization of phosphatidylserine occurs due to the disintegration of the membrane structure, indicating that DK1 induced cell death via apoptosis in both HT29 and SW620 cells. These results are also in agreement with previous studies conducted on breast cancer cells and osteosarcoma cells, which concluded that DK1 induces cell death through the apoptosis pathway [ 15 , 16 ].…”

“…The effects of DK1 treatment towards HT29 and SW620 cells were further assessed by conducting qRT-PCR and human apoptosis proteome profiler. Apoptosis is mainly regulated by the caspase cascade and a family of apoptotic proteins called Bcl-2 family protein [ 16 , 18 ]. Caspase-3, caspase-8, and caspase-9 were chosen as these genes are involved in the caspase cascade that would then trigger the regulation of Bax and cytochrome c. The gene regulation of cyclin A and Cdk 2 genes were also assessed as they form a complex that plays an important role in the cell cycle progression at S phase, as shown in Figure 4 .…”

“…A recent study by Ali et al found that DK1 treatment was more selective and cytotoxic towards MCF-7 and MDA MB-213 breast cancer cell lines thantheMCF-10A normal breast cell line and induced G2/M phase arrest in the breast cancer cells [ 15 ]. Aziz et al reported remarkable results where DK1 induced S phase arrest in cell cycle progression and apoptosis via the intrinsic pathway in two bone cancer cell lines, U-2 OS and MG-63 [ 16 ]. These two studies showed that DK1 may be a potential multi-target drug in cancer therapy.…”

Novel molecular insights and new therapeutic strategies in osteosarcoma

“…Curcumin has been shown to play an anti-cancer effect in OS cells, its mediation can potentially be occurred via Notch-1 signaling inactivation, suggesting that curcumin is involved in upregulation of Notch-1 may be considered as a potential therapeutic strategy for OS [ 191 ]. It has been reported that apoptosis can be induced by Curcumin analog DK1 in human OS cells via mitochondria-dependent signaling, indicating its potential for future cancer treatment [ 192 ]. Moreover, Curcumin-loaded nanoparticles have been revealed to be involved in increasing apoptosis in OS cells [ 193 ].…”

A trienone analog of curcumin, 1,7-bis(3-hydroxyphenyl)-1,4,6-heptatrien-3-one, possesses ROS- and caspase-mediated apoptosis in human oral squamous cell carcinoma cells in vitro

“…Synthetic curcuminoid derivatives have, therefore, been developed by modifying the parent structure to improve anticancer potency. These analogs include DK1 [13], B63 [14], EF24 [15], WZ35 [16], and those with 1,4,6-trien-one function or trienones [17].…”