GO-Y078, a Curcumin Analog, Induces Both Apoptotic Pathways in Human Osteosarcoma Cells via Activation of JNK and p38 Signaling

Abstract

Osteosarcoma is the most common primary bone malignancy in teenagers and continues to confer a generally poor prognosis due to its highly metastatic potential. Poor solubility in water and instability of curcumin limits its bioavailability for use in the adjuvant situation to improve the prognosis and the long-term survival of patients with osteosarcoma. To further obtain information regarding the apoptosis induced by a new curcumin analog, GO-Y078, in human osteosarcoma cells, flow cytometric analysis, annexin V-FITC/PI apoptosis staining assay, human apoptosis array, and Western blotting were employed. GO-Y078 dose-dependently decreased viabilities of human osteosarcoma U2OS, MG-63, 143B, and Saos-2 cells and induced sub-G1 fraction arrest and apoptosis in U2OS and 143B cells. In addition to the effector caspase 3 and poly adenosine diphosphate-ribose polymerase, GO-Y078 significantly activated both initiators of extrinsic caspase 8 and intrinsic caspase 9, whereas cellular inhibitors of apoptosis 1 (cIAP-1) and X-chromosome-linked IAP (XIAP) in U2OS and 143B cells were significantly repressed. Moreover, GO-Y078 increased phosphorylation of extracellular signal-regulated protein kinases (ERK)1/2, c-Jun N-terminal kinases (JNK)1/2, and p38 in U2OS and 143B cells. Using inhibitors of JNK (JNK-in-8) and p38 (SB203580), GO-Y078′s increases in cleaved caspases 8, 9, and 3 could be expectedly suppressed, but they could not be affected by co-treatment with the ERK inhibitor (U0126). Altogether, GO-Y078 simultaneously induces both apoptotic pathways and cell arrest in U2OS and 143B cells through activating JNK and p38 signaling and repressing IAPs. These findings contribute to a better understanding of the mechanisms responsible for GO-Y078′s apoptotic effects on human osteosarcoma cells.

Curcumin analog HO‐3867 triggers apoptotic pathways through activating JNK1/2 signalling in human oral squamous cell carcinoma cells

“…To test this hypothesis, we first examined human apoptosis array (ARY009, R&D systems) in SCC‐9 cells. The human apoptosis array contains 35 proteins that associated with apoptotic process as our previous reports 45 . We identified a serial change in the protein amounts (Figure 4A) We found that cleaved caspase‐3 was increased by approximately 2.3‐fold in HO‐3867‐treated SCC‐9 cells compared to the vehicle control (Figure 4B).…”

“…The human apoptosis array contains 35 proteins that associated with apoptotic process as our previous reports. 45 We identified a serial change in the protein amounts (Figure 4A) We found that cleaved caspase-3 was increased by approximately 2.3-fold in HO-3867-treated SCC-9 cells compared to the vehicle control (Figure 4B). Nevertheless, we also found that XIAP, cIAP and Survivin were reduced by 60% upon HO-3867 (Figure 4B).…”

HO-3867 Induces Apoptosis via the JNK Signaling Pathway in Human Osteosarcoma Cells

“…We can determine the phases of the cell cycle affected by HO-3867, as well as several other cellular components such as DNA, using flow cytometry analysis on U2OS and HOS cells. Summarily, we plated 8 × 105 U2OS and 6 × 10 5 HOS cells in 6 cm dishes and placed them in experimental concentration range (0, 2, 4, 8, and 16 µM) of HO-3867 for 24 h. After propidium iodide (PI) staining, 2 × 105 U2OS and HOS cells were placed in an Eppendorf tube to analyze the cell cycle using a BD AccuriTM C6 Plus personal flow cytometer [28,29].…”

“…Following the protocol given by the manufacturer (BD Biosciences, San Jose, CA, USA), FITC Annexin V Apoptosis Detection Kit I was administered, followed by the analysis of the cell cycle through flow cytometry. Annexin V-FITC apoptosis staining was used in conjugation with PI staining to determine apoptosis from necrosis [29,30].…”

Deoxyshikonin Mediates Heme Oxygenase-1 Induction and Apoptotic Response via p38 Signaling in Tongue Cancer Cell Lines

“…Levels of cellular DNA and membrane annexin V were monitored to determine the cell cycle and apoptotic response of DSK-treated tongue cancer cells, respectively, by using flow cytometry as described previously [42]. In brief, 3.5 × 10 5 /well HSC-3 and 3 × 10 5 /well SCC-9 cells in 6-well plates in response to various concentrations of DSK for 24 h were stained with PI (Invitrogen, Carlsbad, CA, USA), and the analyses of cell cycle distribution were performed with a BD AccuriTM C6 Plus personal flow cytometer (BD Biosciences, San Jose, CA, USA).…”