Thymoquinone Augments Methotrexate-Induced Apoptosis on Osteosarcoma Cells

Abstract

Background Osteosarcoma (OS) as the most frequent primary bone malignancy in children and adolescents has a short survival rate in advanced stages. Alternative herbal medicines with fewer side effects or the potency to protect common therapy’s side effects can be helpful in combinational therapies. Herein, we aim to explore the effects of Thymoquinone (TQ) combined with Methotrexate (MTX) on Saos-2 cells apoptosis. Methods The effects of TQ and MTX alone or in combination on Saos-2 cell viability were measured by MTT assay. Real-time PCR was applied for the measurement of Bax, BCL-2, and caspase-9 mRNA expression. Apoptosis evaluation was conducted by flow cytometry. Results TQ improves the cytotoxic effects of MTX on Saos-2 cells proliferation at lower doses. Indeed, the IC50 of MTX decreased from 26 μM to 15 μM when it combined with TQ. TQ and MTX can induce the expression level of pro-apoptotic factors, Bax and caspase-9 while inhibiting anti-apoptotic protein BCL-2. Moreover, the combination of TQ and MTX potentiates apoptosis to 73%, compared to either TQ (48%) or MTX (53%) treated cells. Conclusion The co-treatment of TQ and MTX is associated with the up-regulation of apoptotic factors and down-regulation of anti-apoptotic factors, conducting apoptosis aggravation and OS cell death.

Thymoquinone induces G2/M cell cycle phase arrest and apoptosis through inhibition of JNK phosphorylation and induction of p53 and p21 expression in HT-1080 fibrosarcoma cells

“…Induction of apoptosis was a central effect of TQ in HT-1080 cells, evidenced by increased the rate of AnnexinV positive cells in early and late stages of the apoptotic process. Our data further con rm the pro-apoptotic properties of TQ reported earlier on solid (31,32) and lymphoid cancer cells (11).…”

“…Our study is the rst to demonstrate that TQ anti-cancer activity towards the human brosarcoma HT1080 cells involve inhibition of JNK phosphorylation, and induction of p53 and p21 expression. The selective toxicity of TQ in cancer treatment was demonstrated in lymphoid and non-lymphoid cancers(46) and non-lymphoid (19,29,31,47). Concerning its potential association with other chemotherapeutic agents, TQ has been shown to synergize with several some drugs such as ifosfamide and cisplatin (30,47).…”

Thymoquinone: A Promising Therapeutic Agent for the Treatment of Colorectal Cancer

“…Particular attention is currently being paid to research into the anticancer effects of TQ. Thymoquinone has anticancer properties that have been confirmed in preclinical studies of many types of cancer, including ovarian, colon, laryngeal, breast, leukemia, lung and osteosarcoma [67][68][69][70][71][72][73][74][75][76]. Currently, there is only one clinical trial evaluating the chemopreventive effect of thymoquinone on potentially malignant oral lesions in the clinicaltrials.gov database (accessed 14 November 2023).…”

Potential anticancer properties and mechanisms of thymoquinone in colorectal cancer

“…A vast variety of proteins and signaling pathways that play key roles in cancer pathogenesis are reported to be modulated by TQ including inhibition of E2F-1 [ 67 ], eEF-2 K [ 21 ], microphthalmia‑associated transcription factor (MITF) [ 33 ], Rac1 [ 40 ], Notch1 [ 68 ], Src/FAK [ 21 , 31 , 69 – 71 ], PI3K/Akt/mTOR [ 21 , 31 , 34 , 36 , 48 , 72 , 73 ], TGF‑β/Smad2/3 [ 32 ], Wnt/β-catenin [ 33 , 42 ], tubulin α/β [ 74 ], NF-κB and p65 [ 21 , 22 , 31 ], TNF-α [ 75 ], anti-apoptotic proteins (IAP1/2, XIAP, Bcl-2, Bcl-xL Mcl-1, c-FLIP and survivin) [ 22 , 30 , 31 , 48 , 50 , 62 , 63 , 76 ], PD-L1 [ 45 , 64 ], HIF-1α [ 77 ], MUC4 [ 69 , 78 ], ENA-78 and Gro [ 79 ], androgen receptor [ 67 ], Plk1 [ 80 ], IRAK1 [ 81 ], proliferative proteins (cyclin A, cyclin B1, cyclin D1/2/3, cyclin E, CDK2/4, c-Myc, Ki-67, PCNA) [ 22 , 31 , 42 , 48 , 50 , 57 , 65 , 67 , 73 , 82 , 83 ], CXCR4 [ 84 , 85 ], Integrin-β1 [ 47 ], Beclin-1 and LC3 [ 47 ], COX-2 [ 22 , 84 ], HSP70 [ 58 ], u-PA [ 86 ], MMP-2/9 [ 22 , 39 , 70 ...…”

“…In-vitro studies also mentioned the combination of TQ with anti-neoplastic agents or flavonoids such as thalidomide [ 31 ], temozolomide [ 110 ], bortezomib [ 31 ], doxorubicin [ 30 ], 5-fluorouracil [ 30 , 111 ], paclitaxel [ 30 ], methotrexate [ 62 , 63 ], cisplatin [ 79 , 112 ], tamoxifen [ 113 ], topotecan [ 114 ], gemcitabine [ 68 , 94 ], curcumin [ 36 ], quercetin [ 115 ], and emodin [ 116 ] augmented cytotoxic effects of these chemicals against cancer cells.…”