Jinzhu Ma1, Yiping Zhu2, Zhen Wang2, Jiawei Zan3, Long Cao1, Zunyong Feng1, Senlin Wang1, Qian Fan4, Liang Yan1. 1. Provincial Key Laboratory of Biological Macromolecules Research, Wannan Medical College, Wuhu 241002, China. 2. 2 First Affiliated Hospital of Wangnan Medical College, Wuhu 241022, China. 3. School of Life Sciences, Anhui Normal University, Wuhu 241002, China. 4. Tianjin Medical University Cancer Institute and Hospital, Tianjin 300060, China.
Abstract
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 (P < 0.05). The results of qRT-PCR showed no significant differences in the expression levels of PLK1 among the cells with different treatments (P > 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.
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 (P &lt; 0.05). The results of qRT-PCR showed no significant differences in the expression levels of PLK1 among the cells with different treatments (P &gt; 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.
Authors: M J Fernández-Aceñero; D Cortés; T Gómez del Pulgar; A Cebrián; L Estrada; J Martínez-Useros; A Celdrán; J García-Foncillas; C Pastor Journal: Pathol Oncol Res Date: 2015-11-17 Impact factor: 3.201