Anti-Metastatic and Anti-Angiogenic Effects of Curcumin Analog DK1 on Human Osteosarcoma Cells In Vitro

Abstract

Osteosarcoma (OS) is a life-threatening malignant bone tumor associated with poor prognosis among children. The survival rate of the patient is still arguably low even with intensive treatment provided, plus with the inherent side effects from the chemotherapy, which gives more unfavorable outcomes. Hence, the search for potent anti-osteosarcoma agent with promising safety profile is still on going. Natural occurring substance like curcumin has gained a lot of attention due to its splendid safety profile as well as it pharmacological advantages such as anti-metastasis and anti-angiogenesis. However, natural curcumin was widely known for its poor cellular uptake, which undermines all potential that it possesses. This prompted the development of synthetically synthesized curcuminoid analog, known as (Z)-3-hydroxy-1-(2-hydroxyphenyl)-3-phenylprop-2- en-1-one (DK1). In this present study, in vitro scratch assay, transwell migration/invasion assay, HUVEC tube formation assay, and ex vivo rat aortic ring assays were performed in order to investigate the anti-metastatic and anti-angiogenic potential of DK1. For further comprehension of DK1 mechanism on human osteosarcoma cell lines, microarray gene expression analysis, quantitative polymerase chain reaction (qPCR), and proteome profiler were adopted, providing valuable forecast from the expression of important genes and proteins related to metastasis and angiogenesis. Based on the data gathered from the bioassays, DK1 was able to inhibit the metastasis and angiogenesis of human osteosarcoma cell lines by significantly reducing the cell motility, number of migrated and invaded cells as well as the tube formation and micro-vessels sprouting. Additionally, DK1 also has significantly regulated several cancer pathways involved in OS proliferation, metastasis, and angiogenesis such as PI3K/Akt and NF-κB in both U-2 OS and MG-63. Regulation of PI3K/Akt caused up-regulation of genes related to metastasis inhibition, namely, PTEN, FOXO, PLK3, and GADD45A. Meanwhile, NF-κB pathway was regulated by mitigating the expression of NF-κB activator such as IKBKB and IKBKE in MG-63, whilst up-regulating the expression of NF-κB inhibitors such as NFKBIA and NFKBIE in U-2 OS. Finally, DK1 also has successfully hindered the metastatic and angiogenic capability of OS cell lines by down-regulating the expression of pro-metastatic genes and proteins like MMP3, COL11A1, FGF1, Endoglin, uPA, and IGFBP2 in U-2 OS. Whilst for MG-63, the significantly down-regulated oncogenes were Serpin E1, AKT2, VEGF, uPA, PD-ECGF, and Endoglin. These results suggest that curcumin analog DK1 may serve as a potential new anti-osteosarcoma agent due to its anti-metastatic and anti-angiogenic attributes.

Develop a Novel Signature to Predict the Survival and Affect the Immune Microenvironment of Osteosarcoma Patients: Anoikis-Related Genes

“…Additionally, compounds like the curcumin analog DK1 can suppress prometastatic genes and proteins including MMP3, thereby reducing the metastatic and angiogenic potential of OS cell lines [36]. Notably, MMP3 has been implicated in stimulating tumor cell metastasis in OS [36].…”

Sanguinarine Inhibition of TNF-α-Induced CCL2, IKBKE/NF-κB/ERK1/2 Signaling Pathway, and Cell Migration in Human Triple-Negative Breast Cancer Cells

“…Our migration analysis suggested the antimetastatic activity of SANG in both cell lines under investigation with relatively higher effects in MDA-MB-231 cells ( Figure 6 ). Angiogenesis is cohesively linked to metastasis, fostering tumor progression at distant sites [ 136 , 137 ]. Metastasis is the key to cancer-related death [ 138 ] since, in the metastasis process, tumor cells segregate from the initial tumor and invade surrounding or distant tissues to establish a secondary tumor [ 48 ].…”

Curcumin and its Analogs and Carriers: Potential Therapeutic Strategies for Human Osteosarcoma

“…In addition to cytotoxic activity in breast MCF-7 cancer cells by inducing G 2 /M cell cycle arrest and apoptosis, DK1 leads to more morphological changes in human osteosarcoma U2OS and MG-63 cells and a significant reduction in cell numbers by inducing apoptosis than curcumin, which occurs through the mitochondrial-dependent signaling pathway [162]. Furthermore, DK1 has been reported to inhibit metastasis and angiogenesis by up-regulation of PI3K/Akt in U2OS cells and NF-κB in both U2OS and MG-63 cells [48]. However, the authors should exclude the effects of IC25 and IC 50 on cell growth before drawing any conclusions about the inhibition of DK1 motility, migration, invasion, and angiogenesis.…”

“…After carcinogenesis, several distinct hallmarks of cancer cells develop, such as evading apoptotic cell death, endless proliferation, and distant metastasis, resulting in host mortality. During tumor growth or disease, a physiological or pathological process, newly formed vascular vessels evolve from pre-existing vessels through angiogenesis [48]. Autophagy (autophagocytosis), a cellular degradation process, allows cells to remove unnecessary or dysfunctional components by inducing the lysosomal recycling of intracellular nutrients and caspaseindependent cell death in conditions of starvation [49].…”