Effects of microRNA-9 on proliferation and migration of thyroid papillary cancer stem cells in vitro

doi:10.3969/j.issn.2095-4344.1763

Abstract

Abstract:

BACKGROUND: MicroRNA-9 (miR-9) exerts different effects in different types of tumors and at different stages of tumorigenesis, but its role in thyroid papillary carcinoma remains unclear.
OBJECTIVE: To analyze the effects of miR-9 on the proliferation and migration of thyroid papillary cancer stem cells in vitro.
METHODS: ALDH1 positive cells and ALDH1 negative cells were separated from human papillary thyroid cancer cells by flow cytometry. Expression of miR-9 mRNA was detected using reverse transcription PCR. ALDH1 positive cells in logarithmic growth phase were transfected with miR-9 mimetic, miR-9 inhibitor or miR mimetic control. After 48 hours, the transfection efficiency of miR-9 was detected by qRT-PCR; cell proliferation, apoptosis and migration were detected by MTT, TUNEL, and Transwell chamber assay, respectively. The expression of apoptosis-related proteins (Bcl-2 and Bax) and migration-related proteins (matrix metalloproteinase-2 and matrix metalloproteinase-9) was detected by western blot.
RESULTS AND CONCLUSION: Successful transfections were confirmed by increased miR-9 gene expression in the miR-9 mimetic group and decreased miR-9 expression in the miR-9 inhibitor group relative to the miR mimetic control group (P < 0.05). Compared with the miR mimetic control group, the miR-9 mimetic group showed decreased cell proliferation and migration ability, and increased apoptosis, while in the miR-9 inhibitor group, cell proliferation and migration abilities were increased, and cell apoptosis decreased. Compared with the miR mimetic control group, the expression of Bcl-2 protein was decreased and the expression of Bax protein increased in the miR-9 mimetic group (P < 0.05), and the expression of Bcl-2 protein increased and expression of Bax protein decreased in the miR-9 inhibitor group (P < 0.05). Compared with the miR mimetic control group, the expression of matrix metalloproteinase 2 and matrix metalloproteinase-9 was decreased in the miR-9 mimetic group (P < 0.05), and the expression of matrix metalloproteinase 2 and matrix metalloproteinase-9 in the miR-9 inhibitor group was increased (P < 0.05). To conclude, miR-9 can inhibit the proliferation and migration of thyroid papillary cancer stem cells in vitro, but increase cell apoptosis.

Key words: papillary thyroid carcinoma, tumor stem cells, ALDH1 positive cells, miR-9, gene transfection, cell proliferation, cell migration, apoptosis

CLC Number:

Han Fei, Qin Haixia. Effects of microRNA-9 on proliferation and migration of thyroid papillary cancer stem cells in vitro[J]. Chinese Journal of Tissue Engineering Research, 2019, 23(21): 3404-3409.

Figures/Tables 6

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