Design and Construction of 3D in Vitro Model for Investigation of Sphingosine-1-Phosphate Role in Ovarian Carcinoma

Despite the major progress in cancer research, most cancer deaths are due to the dissemination of metastases. Ovarian carcinoma presents a significant challenge both in designing effective drug treatment and in understanding cellular processes during disease progression, especially during the transition from a solid form of the tumor to a detached cellular form, in effusion fluid. Emerging evidence shows the various functions of sphingolipids in cellular trafficking and cell motility. Specifically, Sphingolipid-1-phosphate (S1P) has been implicated as a potent regulator of cancer progression. Cancer research is currently limited by the lack of an appropriate in vitro model of the tumor microenvironment. An ideal model must take into account the various components of this microenvironment, including biophysical stimulation, matrix architecture, proper oxygenation and other factors, to better mimic the complex structure and behavior of a solid tumor in vivo. Constructing the tumor cell three-dimensional (3D) microenvironment will provide a better cancer model to investigate the various factors involved and particularly the functions of sphingolipids, especially S1P as regulatory mechanism governing OC progression.

Herein, our main objective was to analyze the mapping of the S1P axis involved genes and with acquired data to establish a 3D in vitro culture system of OC cells, using alginate macroporous-based scaffolds under dynamic conditions to mimic the 3D metastatic tumor microenvironment. Our results indicated that cultivation within alginate porous scaffolds under specific dynamic conditions resulted in S1P receptor mRNA expression levels, relatively close to those of the in vivo samples, compared to monolayer cultures, static scaffold and spheroid cultures. Moreover, cultivation also resulted in receptor distribution, similar to that of the in vivo OC forms.