Biological evaluation of antiproliferative and anti-invasive properties of an androstadiene derivative on human cervical cancer cell lines

Abstract

Gynaecological cancers are leading cause of death: breast cancer is the most frequently diagnosed type of malignancies, and cervical neoplasms rank fourth for both incidence and mortality among women worldwide. In one of our previous studies, favourable antiproliferative and antimetastatic properties of a newly synthesized androstane derivative, 17APAD have been demonstrated on breast cancer cell lines with different expression patterns of hormone receptors. The aim of the current study was to investigate the antitumoral potential of this molecule in cervical cancer cell lines, including SiHa cells positive for human papilloma virus (HPV) type 16 and HPV-negative C33A cells. 17APAD exerted pronounced growth-inhibition (with IC₅₀ values ranging from 0.76 to 1.72 μM with considerable cancer selectivity), while cisplatin used as a reference agent yielded higher IC₅₀ values (ranging from 3.69 to 12.43) and less selectivity, as evidenced by MTT assay. The proapoptotic effect and morphological changes induced by 17APAD were detected by Hoechst 33258-propidium iodide or Annexin V-Alexa488-propidium iodide fluorescent double staining methods, supplemented with a caspase-3 activity assay to identify the mechanism behind the programmed cell death induced by 17APAD. Additionally, significant and concentration-dependent elevation of the ratio of cells in the G2/M phase, on the expense of G0/G1 phase, was observed after 48 h of exposure to 17APAD. Besides its potent antiproliferative properties against both cervical cancer cell lines, 17APAD elicited a remarkable inhibition of cell migration and invasion as detected in wound-healing and Boyden chamber assays, respectively. The mechanisms of action underlying the effects of 17APAD on cell proliferation and motility were independent of androgenic activity, as demonstrated by the Yeast Androgen Screen method. Our results provide new evidence for the proapoptotic and anti-invasive properties of 17APAD, suggesting that it is worth of further research, as a promising prototype for designing novel anticancer agents.

Keywords: Anti-invasive; Antiproliferative; Cervical cancer; Heterocyclic androstane.