The investigation of ceranib-2 on apoptosis and drug interaction with carboplatin in human non small cell lung cancer cells in vitro

Abstract

Ceramide is found to be involved in inhibition of cell division and induction of apoptosis in certain tumour cells. Ceranib-2 is an agent that increases ceramide levels by inhibiting ceramidase in cancer cells. Therefore, we aimed to investigate the effects of ceranib-2 on cell survival, apoptosis and interaction with carboplatin in human non-small cell lung cancer cells. The cytotoxic effect of ceranib-2 (1-100 µM) was determined by MTT assay in human lung adenocarcinoma (A549) and large cell lung carcinoma (H460) cells. Carboplatin (1-100 µM) and lung bronchial epithelial cells (BEAS-2B) were used as positive controls. Morphological and ultrastructural changes were analysed by light microscope and TEM. Apoptotic/necrotic cell death and acid ceramidase activity were analysed by ELISA. Combination effects of ceranib-2 and carboplatin were investigated by MTT. The expression levels of CASP3, CASP9, BAX and BCL-2 were examined by qRT-PCR. The IC of ceranib-2 was determined as 22 μM in A549 cells and 8 μM in H460 cells for 24 h. Morphological changes and induction of DNA fragmentation have revealed apoptotic effects of ceranib-2 in both cell lines. Ceranib-2 and carboplatin has shown synergism in combined treatment at 10 and 25 μM doses in H460 cells for 24 h. Ceranib-2 inhibited acid ceramidase activity by 44% at 25 µM in H460 cells. Finally, CASP3, CASP9 and BAX expressions were increased while BCL-2 expression was reduced in both cells. Our results obtained some preliminary results about the cytotoxic and apoptotic effects of ceranib-2 for the first time in NSCLC cell lines.

Inhibitors of Ceramide- and Sphingosine-Metabolizing Enzymes as Sensitizers in Radiotherapy and Chemotherapy for Head and Neck Squamous Cell Carcinoma

“…A novel CDase inhibitor, Ceranib-2, inhibited CDase activity, increased ceramide levels, reduced S1P levels, and inhibited cell proliferation and tumor growth [118]. Ceranib-2 and carboplatin exhibit synergism in combination for non-small cell lung cancer, where caspase 3, caspase 9, and Bax expression were increased, whereas Bcl-2 expression was reduced [122]. Treatment of acute myeloid leukemia cell lines with a novel aCDase inhibitor, SACLAC, effectively blocked aCDase activity and induced a decrease in S1P and an increase in total ceramide levels [123].…”

One‐Step Microfluidic Coating of Phospholipid Microbubbles with Natural Alginate Polymer as a Delivery System for Human Epithelial Lung Adenocarcinoma

“…First, cells in exponential growth phase were counted by using trypan blue dye in inverted light microscope (Axio Vert.A1, Carl Zeiss, Germany) and 1 × 10 4 cells per well were cultured in 96 well plates at 37 °C, 5% CO 2 in a fully humidified incubator overnight. After the culture medium was removed, carboplatin (100 µg mL −1 ) [ 8,9 ] was added to the cells in total volume of 360 µL for the elimination of air space when the plates reversed. Also, required CAMBs volumes were calculated according to carboplatin concentration equally and MBs, AMBs volumes were used equal to CAMBs.…”

“…[ 8 ] One commonly used NSCLC cell line is A549, an epithelial carcinoma, known to be KRAS mutant. [ 9 ] Despite intensive studies on lung cancer, the treatment of this disease has not yet been successful due to the serious side effects of the chemotherapeutics used and drug resistance. Therefore, there is a need to investigate new therapeutic approaches such as new drug delivery systems, [ 10 ] which are compatible.…”

Elusive Roles of the Different Ceramidases in Human Health, Pathophysiology, and Tissue Regeneration

“…The activation of ACDase induces a pro-survival state, while its inhibition leads to cell death through a variety of apoptotic pathways mediated by caspases (CASP), poly (adp-ribose) polymerase (PARP), or cathepsins (CTS) [ 10 , 17 , 18 , 19 , 20 , 21 , 22 , 23 , 24 , 25 ]. Cathepsin B and Cathepsin D are activated during ceramide-induced apoptosis but are inhibited by ACDase activity [ 19 , 25 ].…”