Cyclooxygenase-2 inhibitor is a robust enhancer of anticancer agents against hepatocellular carcinoma multicellular spheroids

Abstract

PurposeCelecoxib, an inhibitor of cyclooxygenase-2 (COX2), was investigated for enhancement of chemotherapeutic efficacy in cancer clinical trials. This study aimed to determine whether celecoxib combined with 5-fluorouracil or sorafenib or gefitinib is beneficial in HepG2 multicellular spheroids (MCSs), as well as elucidate the underlying mechanisms.MethodsThe human hepatocellular carcinoma cell line HepG2 MCSs were used as in vitro models to investigate the effects of celecoxib combined with 5-fluorouracil or sorafenib or gefitinib treatment on cell growth, apoptosis, and signaling pathway.ResultsMCSs showed resistance to drugs compared with monolayer cells. Celecoxib combined with 5-fluorouracil or sorafenib exhibited a synergistic action. Exposure to celecoxib (21.8 μmol/L) plus 5-fluorouracil (8.1 × 10−3 g/L) or sorafenib (4.4 μmol/L) increased apoptosis but exerted no effect on COX2, phosphorylated epidermal growth-factor receptor (p-EGFR) and phosphorylated (p)-AKT expression. Gefitinib (5 μmol/L), which exhibits no growth-inhibition activity as a single agent, increased the inhibitory effect of celecoxib. Gefitinib (5 μmol/L) plus celecoxib (21.8 μmol/L) increased apoptosis. COX2, p-EGFR, and p-AKT were inhibited.ConclusionCelecoxib combined with 5-fluorouracil or sorafenib or gefitinib may be superior to single-agent therapy in HepG2 MCSs. Our results provided molecular evidence to support celecoxib combination-treatment strategies for patients with human hepatocellular carcinoma. MCSs provided a good model to evaluate the interaction of anticancer drugs.

Parecoxib prevents complications in hepatocellular carcinoma patients receiving hepatic transarterial chemoembolization: a prospective score-matched cohort study

“…COX-2 is overexpressed in HCC and is thought to contribute to hepatocarcinogenesis [ 22 ]. Consistent with that idea, inhibition of COX-2 reportedly enhances chemotherapeutic efficacy in preclinical research and cancer clinical trials [ 23 , 24 ]. Although there was no difference between the effect of TACE on tumors in the parecoxib and control groups in our study, we can draw no definite conclusion yet, since this is a short-term medication.…”

Comparison of NSAIDs activity in COX-2 expressing and non-expressing 2D and 3D pancreatic cancer cell cultures

“…EC 50 value of celecoxib in 3D cell culture was 60.6±15.9 μM whereas in 2D cell culture it was 31.0±12.4 μM. 31 The reason behind these differences of drug activity between 2D and 3D cell cultures might be distinct mechanism of actions.…”

β1 integrin-mediated multicellular resistance in hepatocellular carcinoma through activation of the FAK/Akt pathway

“…HepG2 cells and Bel-7402 cells (American Type Culture Collection, Manassas, VA, USA) were cultured in RPMI-1640 medium containing 10% fetal bovine serum in 5% CO 2 at 37°C as a monolayer culture. As previously described, 10 – 12 a liquid overlay technique was used to obtain tumour multicellular spheroids. Briefly, a cell suspension was seeded at 2 x 10 5 cells in each culture flask coated with 2% agarose (Sigma-Aldrich, St Louis, MO, USA) before cell plating.…”

“…The cell proliferation inhibition effects of 5-FU and CDDP in monolayer cells and multicellular spheroids were determined using a 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay as previously described. 10 – 12 For each well, a total of 2500 cells were seeded as either monolayer cells or multicellular spheroids in 150 μl of tissue culture medium. Next, the cells were treated with 5-FU (Xudong Haipu Pharmaceutical Company, Shanghai, China) and CDDP (Qilu Pharmaceutical Company, Shandong, China) for 48 h at the indicated concentrations.…”

“… 3 , 5 – 7 Previous studies have shown that tumour spheroids were generally more resistant to chemotherapy than their conventional 2D monolayer culture counterparts. 8 – 12 This novel concept, called multicellular resistance, emphasized the importance of utilizing tumour multicellular spheroids for the evaluation of anticancer drug and mechanistic studies. 13 …”