1.miR-593 inhibits proliferation of colon cancer cells by down-regulating PLK1.
Jinzhu MA ; Yiping ZHU ; Zhen WANG ; Jiawei ZAN ; Long CAO ; Zunyong FENG ; Senlin WANG ; Qian FAN ; Liang YAN
Journal of Southern Medical University 2019;39(2):144-149
OBJECTIVE:
To explore the role of miR-593 in regulating the proliferation of colon cancer cells and the molecular mechanism.
METHODS:
Bioinformatics analysis identified PLK1 as the possible target gene of miR-593. Luciferase assay was employed to verify the binding between miR-593 and PLK1, and qRT-PCR and Western blotting were used to verify that PLK1 was the direct target gene of miR-593. CCK-8 assay was performed to test the hypothesis that miR-593 inhibited the proliferation of colon cancer cells by targeting PLK1.
RESULTS:
Luciferase assay identified the specific site of miR-593 binding with PLK1. Western blotting showed a significantly decreased expression of PLK1 in the colon cancer cells transfected with miR-593 mimics and an increased PLK1 expression in the cells transfected with the miR-593 inhibitor as compared with the control cells ( < 0.05). The results of qRT-PCR showed no significant differences in the expression levels of PLK1 among the cells with different treatments ( > 0.05). The cell proliferation assay showed opposite effects of miR-593 and PLK1 on the proliferation of colon cancer cells, and the effect of co-transfection with miR-593 mimic and a PLK1-overexpressing plasmid on the cell proliferation was between those in PLK1 over-expressing group and miR-593 mimic group.
CONCLUSIONS
miR-593 inhibits the proliferation of colon cancer cells by down-regulating PLK1 and plays the role as a tumor suppressor in colon cancer.
Binding Sites
;
Cell Cycle Proteins
;
genetics
;
metabolism
;
Cell Line, Tumor
;
Cell Proliferation
;
Colonic Neoplasms
;
metabolism
;
pathology
;
Down-Regulation
;
Gene Expression Regulation, Neoplastic
;
Genes, Tumor Suppressor
;
Humans
;
In Vitro Techniques
;
MicroRNAs
;
genetics
;
metabolism
;
Protein-Serine-Threonine Kinases
;
genetics
;
metabolism
;
Proto-Oncogene Proteins
;
genetics
;
metabolism
;
Reverse Transcriptase Polymerase Chain Reaction
;
Sincalide
;
metabolism
;
Transfection