MicroRNA-200c overexpression in cancer-associated fibroblasts decreases interleukin-2 secretion

Abstract

miR-200c-3p is demonstrated to play the role of tumour suppressor in different tumours. However, the miR-200c-3p biological function in normal fibroblast (NF) and cancer-associated fibroblast (CAF) remains unclear. This investigation aims to study the regulatory role of miR-200c-3p in the secretion of Interleukin-2 (IL-2) in CAF and NF. CAFs and NFs were isolated from tumour and normal tissue specimens respectively. Immunocytochemistry was used to confirm the presence of a fibroblast specific marker, alpha-actin smooth muscle, in NFs and CAFs. NF and CAF were transfected with scramble and miR-200c-3p utilizing the lipofectamine 2000 reagent. The protein levels of IL-2 were measured in CAFs, NFs, and transfected groups with miR-200c-3p and scrambled using an IL-2 enzyme-linked immunoassay kit. miR-200c decreased secretion of IL-2 in transfected CAF and NF compared to controls. Results elucidated that transfection of MiR-200c-3p can decrease the IL-2 secretion and consequently reduce IL-induced tumourigenic manner in the CAF. MiR-200c-3p transfection decreases the Interleukin-2 (IL-2) secretion level, resulting in a reduction in IL-induced tumourigenic manner of cancer-associated fibroblast. image