MiR-216b-5p and HDAC8 Inhibit Breast Cancer Cell Proliferation, Metastasis and Invasion
MiR-216b-5p and HDAC8 Inhibit Breast Cancer Cell Proliferation, Metastasis and Invasion
Qi Zhuo, Yuanchun Yao, Meisongzhu Yang*, Jinhua Chen and Miao Tian
ABSTRACT
The objective of this study was to investigate the mechanism of MiR-216b-5p and HDAC8 inhibiting the proliferation, metastasis and invasion of breast cancer cells. Tissue samples (28) were collected, and malignant and normal tissues were confirmed pathologically. The breast cancer cell line MDAMB-231 and the immortalized normal breast epithelial cell line MCF-10A were used in this study. Western blot analysis of miR-216b-5p and HDAC8 protein expression in tissue samples; reverse transcription quantitative PCR (RT-qPCR) real-time analysis of miR-216b-5p, HDAC8 mRNA expression in cell lines; MTT detection of cell proliferation; wound healing test and transwell assay were used to evaluate the ability of breast cancer cells to metastasize and invade; colony formation test was conducted to detect the effect of HDAC8 on the cells. We found that the expression of miR-216b-5p protein in breast cancer group was lower than that in healthy control group (P<0.05); the expression of HDAC8 protein in the group was higher than that in healthy control group (P<0.05). The MDA-MB-231 group had lower miR-216b-5p mRNA expression than the MCF-10A group (P<0.05), and the MDA-MB-231 group had higher HDAC8 mRNA expression than the MCF-10A group (P<0.05). At 24th h after cell line culture The cell proliferation of miR-216b-5p inhibitor group was higher than that of miR-216b-5p mimic group and control group (P<0.05). At 48th h after cell line culture, the cell proliferation of miR-216b-5p mimic group was lower than that of control group and miR-216b-5p inhibitor group (P<0.05). The cell proliferation of control group was lower than that of the miR-216b-5p inhibitor group (P<0.05). At 72th h after cell line culture, the miR-216b-5p mimic group had lower cell proliferation than miR-216b-5p inhibitor group (P<0.01). And the control group had higher cell proliferation than miR-216b-5p mimic group (P<0.05). The miR-216b-5p inhibitor group had more cell metastasis and invasion than the miR-216b-5p mimic group and the control group (P<0.05). The miR-216b-5p mimic group had less cell metastasis and invasion than the miR-216b-5p inhibitor group and the control group (P<0.05). Compared with the control group and the empty vector group, the HDAC8 gene knockout group had less colony formation (P<0.05). Compared with the miR-216b-5p mimic group, the expression of HDAC8 protein and HDAC8 mRNA increased in the miR-216b-5p inhibitor group (P<0.05). The miR-216b-5p mimic group had lower HDAC8 protein expression and HDAC8 mRNA expression than the control group(P<0.05). It is concluded that HDAC8 is an oncogene that can promote the proliferation and development of breast cancer. MiR-216b-5p has antitumor effect on HDAC8 and can inhibit the proliferation and development of breast cancer.
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