*5.7. ELISA Assay*

As mentioned earlier, spheroids' spent media were collected for collagen invasion assay. Human MMP-9 (ab100610) and MMP-13 (ab100605) were quantified by using specific ELISA kits following the manufacturer's instructions. ELISA Kits were purchased from Abcam Company (Cambridge, UK).

#### *5.8. Human Cytokine Array Assay*

The expression levels of chemokines and cytokines were analyzed using a Human Cytokine Antibody Array (C5) (RayBiotech, Inc., Norcross, GA, USA). As per the manufacturer's instructions, antibody-embedded membranes were incubated with 1 mL of conditioned media (CM) at 4 ◦C overnight, followed by incubation with a biotin-conjugated detection antibody cocktail and diluted HRP-streptavidin at room temperature. Proteins were then visualized using a chemiluminescent substrate reagent. The intensity of each spot represents cytokines, which were then quantified densitometrically using ImageJ software.
