Berberine inhibited metastasis through miR-145/MMP16 axis in vitro

Abstract

Ovarian cancer is the first leading cause of death in gynecological cancers. The continuous survival and metastasis of cancer cells are the main causes of death and poor prognosis in patients with ovarian cancer. Berberine is an effective component extracted from the rhizomes of coptis chinensis and phellodendron chinensis. In our study, we aim to explore the molecular mechanism underlying the regulation of proliferation, migration and invasion by berberine in ovarian cancer cells. CCK8 assay was used for detection of proliferative capacity of SKOV3 and 3AO cells. Wound healing assay was used to estimate cell migration and transwell assay was used to assess cell invasion. The mRNA expression of miR-145 and MMP16 were examined by quantitative real-time polymerase chain reaction (qRT-PCR). The protein level of MMP16 was detected by western blot analysis. In addition, luciferase reporter assays were used to demonstrate MMP16 was a target of miR-145. The results demonstrated berberine inhibited proliferation, migration and invasion, promoted miR-145 expression, and decreased MMP16 expression in SKOV3 and 3AO cells. MMP16 was a target of miR-145. Moreover, downregulation of MMP16 contributed to the inhibition of proliferation, migration and invasion by berberine. Together, our results revealed that berberine inhibited proliferation, migration and invasion through miR-145/MMP16 in SKOV3 and 3AO cells, highlighting the potentiality of berberine to be used as a therapeutic agent for ovarian cancer.

Keywords: Berberine; MMP16; Ovarian cancer; miR-145.

Fig. 1

Berberine promotes miR-145 expression in ovarian cancer cells. a CCK8 assay results showed that berberine could inhibit the growth of SKOV3 and 3AO cells in a dose-dependent manner. b qRT-PCR results showed berberine increased miR-145 expression. P < 0.05, **P < 0.01, t-test

Fig. 2

Berberine inhibits proliferation, migration and invasion through miR-145 in SKOV3 and 3AO cells. a qRT-PCR results showed berberine promoted miR-145 expression, and this promotion effect was offset by transfection of miR-145 inhibitor. b CCK8 assay results showed berberine inhibited proliferation of SKOV3 and 3AO cells, and this inhibition was blocked by downexpression of miR-145. c Wound healing assay showed berberine inhibited migration of SKOV3 and 3AO cells, and this inhibition was blocked by downexpression of miR-145. d Transwell assay showed berberine inhibited invasion of SKOV3 and 3AO cells, and this inhibition was blocked by downexpression of miR-145(200×). e qRT-PCR results showed berberine inhibited the expression of MMP16, and the inhibition of MMP16 was counteracted by downexpression of miR-145. f Western blot assay showed berberine inhibited the expression of MMP16, and the inhibition of MMP16 was counteracted by downexpression of miR-145. P < 0.05, **P < 0.01, t-test

Fig. 3

MMP16 is a target of miR-145. a qRT-PCR results showed overexpression of miR-145 inhibited MMP16 in SKOV3 and 3AO cells. b Western blot results showed overexpression of miR-145 inhibited MMP16 in SKOV3 and 3AO cells. c The luciferase reporter assay showed miR-145 directly. P < 0.05, **P < 0.01, t-test

Fig. 4

miR-145 inhibits proliferation, migration and invasion of SKOV3 and 3AO cells by targeting MMP16. a qRT-PCR results showed overexpression of miR-145 inhibited MMP16 expression, and MMP16 expression was increased by transfection MMP16 plasmid. b Western blot results showed overexpression of miR-145 inhibited MMP16 expression, and MMP16 expression was increased by transfection MMP16 plasmid. c CCK8 assay results showed miR-145 inhibited proliferation, and the inhibition effect was reversed by MMP16 overexpression. d Wound healing assay showed miR-145 inhibited migration, and the inhibition effect was reversed by MMP16 overexpression. e Transwell assay showed miR-145 inhibited invasion, and the inhibition effect was reversed by MMP16 overexpression(200×). P < 0.05, **P < 0.01, t-test

Fig. 5

Schematic representation of the mechanism for berberine‑regulated ovarian cancer progression