9-PAN promotes tubulin- and ROS-mediated cell death in human triple-negative breast cancer cells

Abstract

Objectives To examine the antiproliferative effect of a rationally designed, novel noscapine analogue, 9-((perfluorophenyl)methylene) aminonoscapine, '9-PAN') on MDA-MB-231 breast cancer cell line, and to elucidate the underlying mechanism of action. Methods The rationally designed Schiff base-containing compound, 9-PAN, was characterized using IR, NMR and mass spectra analysis. The effect of the compound on cell viability was studied using an MTT assay. Cell cycle and cell death analyses were performed using flow cytometry. Binding interactions of 9-PAN with tubulin were studied using spectrofluorometry. Reactive oxygen species (ROS) generation and mitochondrial membrane potential (MMP) were investigated using the probes, DCFDA and rhodamine-123, respectively. Immunofluorescence imaging was used to visualize cellular microtubules. Key findings 9-PAN inhibited cell proliferation (IC 50 of 20 AE 0.3 µM) and colony formation (IC 50 , 6.2 AE 0.3 µM) by arresting the cells at G 2 /M phase of the cell cycle. It bound to tubulin in a concentration-dependent manner without considerably altering the tertiary conformation of the protein or the polymer mass of the microtubules in vitro. The noscapinoid substantially damaged cellular microtubule network and induced cell death, facilitated by elevated levels of ROS. Conclusions 9-PAN exerts its antiproliferative effect by targeting tubulin and elevating ROS level in the cells.

Mechanistic insights into encapsulation and release of drugs in colloidal niosomal systems: biophysical aspects

“…After incubation of the cells for a period of three days, the cell viability was determined using an MTT assay. 43 Briefly, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) (0.5 mg ml −1 final concentration) was added to each well and incubated for 4 h at 37 °C. The MTT-containing media was then discarded, and 100 μL of dimethylsulfoxide (DMSO) was added to each well and incubated for 10 min.…”

“…Aer incubation of the cells for a period of three days, the cell viability was determined using an MTT assay. 43 Briey, MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) (0.5 mg ml À1 nal concentration) was added to each well and incubated for 4 h at 37 C. The MTT-containing media was then discarded, and 100 mL of dimethylsulfoxide (DMSO) was added to each well and incubated for 10 min. The absorbance of the samples was measured at 570 nm in a Tecan multimode reader (TECAN innite 200 PRO, Tecan, Switzerland) and the percentage cell viability was calculated as described elsewhere.…”

Noscapine–Amino Acid Conjugates Suppress the Progression of Cancer Cells

“…Over the past two decades, many research groups have developed several analogues of noscapine by incorporating several moieties in the molecule. In recent reports, 9-((perfluoro­phenyl)­methylene)­amino­noscapine (9-PAN) (IC 50 = 20 ± 0.3 μM), N -propargyl­noscapine (NPN) (IC 50 = 1.35 ± 0.2 μM), and 1,3-diynyl derivatives of noscapine (IC 50 = 6.23 μM) have shown excellent results against various breast cancer cell lines by arresting the cells at the G2/M phase of the cell cycle. − 1,3-Benzodioxole-modified noscapinoids also showed a wide range of activity against various cancer lines, particularly MCF-7, with IC 50 = 0.6 ± 0.17 μM . Also, 9-ethynyl noscapinoids induced G2/M arrest and apoptosis by disrupting tubulin polymerization in cervical cancer .…”