The Influence of Antitumor Unsymmetrical Bisacridines on 3D Cancer Spheroids Growth and Viability

Abstract

The culture of 3D spheroids is a promising tool in drug development and testing. Recently, we synthesized a new group of compounds, unsymmetrical bisacridines (UAs), which exhibit high cytotoxicity against various human cell lines and antitumor potency against several xenografts. Here, we describe the ability of four UAs—C-2028, C-2041, C-2045, and C-2053—to influence the growth of HCT116 and H460 spheres and the viability of HCT116 cells in 3D culture compared with that in 2D standard monolayer culture. Spheroids were generated using ultra-low-attachment plates. The morphology and diameters of the obtained spheroids and those treated with UAs were observed and measured under the microscope. The viability of cells exposed to UAs at different concentrations and for different incubation times in 2D and 3D cultures was assessed using 7-AAD staining. All UAs managed to significantly inhibit the growth of HCT116 and H460 spheroids. C-2045 and C-2053 caused the death of the largest population of HCT116 spheroid cells. Although C-2041 seemed to be the most effective in the 2D monolayer experiments, in 3D conditions, it turned out to be the weakest compound. The 3D spheroid culture seems to be a suitable method to examine the efficiency of new antitumor compounds, such as unsymmetrical bisacridines.

c-Myc Protein Level Affected by Unsymmetrical Bisacridines Influences Apoptosis and Senescence Induced in HCT116 Colorectal and H460 Lung Cancer Cells

“…In our Department, we have developed a new group of compounds, unsymmetrical bisacridines (UAs), which exhibited high cytotoxicity against numerous human cancer cells and antitumor activity against several xenografts [ 3 ]. Additionally, UAs turned out to be very potent in inhibiting the growth of HCT116- and H460-derived spheroids [ 27 ]. Moreover, some of the UAs were conjugated with quantum dots (QDs), which improved their delivery to cancer cells [ 28 , 29 , 30 ].…”

“…These observations suggest that C-2041 differed in its way of cellular action from other UAs and that cells treated with this derivative underwent more nonspecific cell death. Here, it is worth to mention that in studies concerning the influence of UAs on 3D spheroids cultures, C-2041 was the compound that inhibited the growth of both HCT116 and H460 spheroids to the least extent [ 27 ]. Another noteworthy observation is that C-2053 was the derivative, which was the least active against HCT116 colon cells, but one of the most potent ones against H460 lung cancer cells.…”

“…Another noteworthy observation is that C-2053 was the derivative, which was the least active against HCT116 colon cells, but one of the most potent ones against H460 lung cancer cells. Moreover, in 3D culture, C-2053 inhibited the growth of spheroids to the greatest extent, especially in the case of HCT116 cells [ 27 ]. The colony-forming assay also confirmed that C-2045 and C-2053 compounds were the most effective derivatives among UAs—cells of both HCT116 and H460 cell lines, after drug treatment, did not return to proliferation.…”

Acid–Base Equilibrium and Self-Association in Relation to High Antitumor Activity of Selected Unsymmetrical Bisacridines Established by Extensive Chemometric Analysis

“…Notably, the compounds which displayed the highest activity strongly inhibited pancreatic cancer cell lines [3]. Studies on the biological effects of these compounds demonstrated their ability to suppress 3D cancer spheroid growth [4]. Moreover, their anticancer activity was enhanced when bound to quaternary quantum dots, resulting in selective upregulation of their cellular uptake [5][6][7].…”

Extensive chemometric analysis exploring the acid-base equilibrium and self-association of unsymmetrical bisacridines reveals characteristics possibly relevant for their anticancer activity

“…UAs exhibited high cytotoxic activity against all 14 tested cancer cell lines as well as antitumor activity after screening in 10 human tumor xenografts in nude mice and against Walker 256 adenocarcinoma [3]. Studies on biological effects of these compounds demonstrated their ability to in uence 3D cancer spheroid growth [4]. Moreover, their anticancer activity was improved when bound to quaternary quantum dots, which selectively up-regulate cellular uptake of these compounds [5][6][7].…”