MicroRNA-195-5p inhibits malignant progression of gallbladder cancer by regulating Wnt3a.

Abstract

We aimed at investigating whether microRNA-195-5p inhibits the malignant proliferation of gallbladder cancer (GBC) via regulating Wnt3a; meanwhile, its relationship with the clinicopathological parameters and prognosis of patients with GBC was also explored. In this study, the tumor tissues and adjacent tissues of 47 GBC patients were tested for microRNA-195-5p expression level by real-time quantitative polymerase chain reaction (qPCR); the relationship between microRNA-195-5p expression and clinical indicators of GBC patients was further analyzed. Control group (NC mimic or NC inhibitor), microRNA-195-5p overexpression group (microRNA-195-5p mimic), and microRNA-195-5p knockdown group (microRNA-195-5p inhibitor) were set in GBC cell lines, respectively. In GBC cell lines GBC-SD and NOZ, cell counting kit-8 (CCK-8), plate cloning experiments and flow cytometry were carried out to assess microRNA-195-5p's effect on proliferation and apoptosis of GBC cells. Further, the interaction between microRNA-195-5p and its downstream gene Wnt3a was explored through Luciferase reporting assay. Our data showed that microRNA-195-5p expression in tumor tissues of GBC patients was remarkably lower than that in adjacent ones. In comparison to patients in highly expressed microRNA-195-5p group, those in lowly expressed microRNA-195-5p had more advanced pathological stage and larger tumor size. Overexpression of microRNA-195-5p markedly attenuated the proliferation capacity of GBC cells as compared to the NC mimic group; in contrast, knockdown of microRNA-195-5p enhanced GBC cell proliferation function of GBC cells in comparison to NC inhibitor group. At the same time, the opposite tendency in cell apoptosis was observed in the above four groups. In GBC tissue specimens, Wnt3a showed an increased expression, which was negatively correlated with microRNA-195-5p. Meanwhile, Luciferase assay verified a binding relationship between microRNA-195-5p and Wnt3a. In addition, we found that over-expressing Wnt3a counteracted the influence of upregulation of microRNA-195-5p on proliferation and apoptosis of GBC cells and thus modulate the malignant progress of GBC cells. In summary, the above studies suggest that low expression of microRNA-195-5p is remarkably relevant to pathological stage and tumor size of patients with GBC. In addition, microRNA-195-5p may suppress the malignant progression of GBC through down-regulating Wnt3a.