The curcumin analog (PAC) suppressed cell survival and induced apoptosis and autophagy in oral cancer cells

Abstract

PAC (3,5-Bis (4-hydroxy-3-methoxybenzylidene)-N-methyl-4-piperidone), a novel bioactive curcumin analog, has been reported to have anticancer properties against various tumors. However, the anti-cancer effects of PAC on oral cavity squamous cell carcinoma were not studied yet. Our aim is to investigate the anti-oral cancer properties of PAC in vitro, and determine the molecular mechanisms underlying these effects. Viability assays including MTT and LDH were conducted to measure cell proliferation. Flow cytometry-based cytotoxicity assay was performed to detect autophagic cell death and oxidative stress markers. Western blotting was used for measuring protein expression/activation in apoptotic, autophagic and pro-carcinogenic cellular signaling pathways. We demonstrated that PAC preferentially and, in a dose, -dependent way kills oral cancer cells, but was not toxic to normal human gingival cells. PAC destabilizes cell-cycle distributions, inhibits the expression of oncogenes (cyclin D1) and that of cyclin-dependent kinase inhibitor (p21WAF1) is upregulated, increases the expression of p53 gene, and inhibits epithelial-mesenchymal transition markers in oral cancer cells. The PAC effect involve various signaling pathways including NF-κB, MAPK, Wnt, caspase-3/9 and PARP1. Finally, PAC demonstrated ability to induce autophagy, decrease production of reactive oxygen species, increase intracellular glutathione (GSH) activity, and reduce mitochondrial membrane potential in oral cancer cells. In conclusion, PAC inhibits the proliferation and increases the apoptosis and autophagy and oxidative stress of oral cancer cells. These effects involve ERK1/2, p38/JNK, NF-κB and Wnt cellular signaling pathways. Overall, our study suggests the potential use of PAC to treat oral cancer.

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

“…70 Interestingly, several analogs of curcumin are identified their antitumour activity through induction of apoptosis in OSCC. 71,72 In HSC-…”

“…Moreover, curcumin promoted apoptosis by inducing cleaved caspase 3 and cleaved PARP in YD10B OSCC cells at the dose of 10 µM 70 . Interestingly, several analogs of curcumin are identified their antitumour activity through induction of apoptosis in OSCC 71,72 . In HSC‐4 and HSC‐2 human oral cancer cells lines, Dibenzylideneacetone inhibits cell viability by triggering apoptosis at the dose of 5–10 µM 71 .…”

Magnolol Triggers Caspase-Mediated Apoptotic Cell Death in Human Oral Cancer Cells through JNK1/2 and p38 Pathways

“…Thus, the role of magnolol-induced ERK1/2 activation in oral cancer requires further study. Moreover, several studies have revealed that JNK1/2 plays an anti-tumor role in oral cancer and emphasizes its positive role in apoptosis [34][35][36]. Numerous studies have also reported that magnolol inhibits cancer progression [14] or induces apoptosis [13,37] by regulating multiple cellular signaling pathways.…”

Naproxen-Loaded Poly(2-hydroxyalkyl methacrylates): Preparation and Drug Release Dynamics

“…The human oral cancer cell line Ca9–22 cells were cultured at 37 °C and 5% CO 2 in RPMI-1640 medium (Thermo Fisher Scientific, Burlington, ON, Canada), supplemented with L-glutamine, 5% fetal bovine serum (FBS, Gibco) provided by the same company, and 1% penicillin/streptomycin solution (Sigma-Aldrich, Oakville, ON, Canada). Cell proliferation was bi-evaluated using two MTT and LDH tests as described by Semlali et al 2021 [ 35 , 36 ] and Contant et al 2021 [ 37 ]. For the MTT assay, 10 5 Ca9-22 cells per well containing the sample were seeded in 24-well plates for 24 h. After adhesion and growth of the cells, the culture medium is replaced by a new one containing a solution of MTT at 5 mg∙ml −1 in PBS and left for 3 h at 37 °C in the dark.…”

“…LDH assay was realized by the LDH Cytotoxicity Detection Kit from Roche, which allows to directly quantify the cell death in culture based on the measurement of lactate dehydrogenase released into growth media. As described in our previous work [ 35 , 36 ], 10 5 cells per well were seeded in 24-well plates containing NPX/pHEMA and NPX/pHPMA with different compositions. After adhesion for 24 h, 50 μL of each supernatant was transferred in triplicate into a 96-well plate and supplemented with 50 μL reconstituted substrate mixture.…”