The effect of piceatannol on the migration and invasion of cervical cancer cells by regulating miR-106b-5p/RUNX3 axis

doi:10.11958/20221529

Abstract

Abstract:

Objective To explore the effect of piceatannol (PIC) on migration and invasion of cervical cancer (CC) cells by regulating microRNA-106b-5p (miR-106b-5p)/RUNT-related transcription factor 3 (RUNX3) axis. Methods Firstly, human CC cells Hela were treated with culture media of different concentrations of PIC (0, 20, 40, 80 and 160 μmol/L), and the effect of PIC on cell proliferation was detected by CCK-8 assay to determine the optimal concentration of PIC. Hela cells were divided into the control group, the PIC group, the PIC+NC mimics group, the PIC+ miR-106b-5p mimics group, the NC inhibitor group, the miR-106b-5p inhibitor group, the miR-106b-5p inhibitor+si-RNA group and the miR-106b-5p inhibitor+si-RUNX3 group. qRT-PCR was used to detect the expression level of miR-106b-5p in Hela cells in each group. Transwell method was used to detect the migration and invasion abilities of Hela cells in each group. Western blot assay was used to detect the protein levels of RUNX3, MMP2 and MMP9 in Hela cells of each group. Dual luciferase reporter gene experiment was used to detect the targeting relationship between miR-106b-5p and RUNX3. Results The proliferation activity of Hela cells decreased with the increase of PIC treatment concentration (P<0.05). The inhibitory effect of 80 μmol/L PIC on Hela cells was nearly to half inhibitory concentration (IC₅₀), so 80 μmol/L was selected as the PIC concentration for subsequent study. Compared with the control group, the expression levels of miR-106b-5p, MMP2 and MMP9 decreased in the PIC group, the numbers of migrating and invasive cells decreased, and the expression level of RUNX3 increased (P<0.05). Compared with the PIC+NC mimics group, the expression levels of miR-106b-5p, MMP2 and MMP9 increased in the PIC+miR-106b-5p mimics group, numbers of migrating and invasive cells increased, and the expression level of RUNX3 decreased (P<0.05). Double luciferase reporter gene assay confirmed RUNX3 as the target gene of miR-106b-5p. Compared with the NC inhibitor group, the expression level of RUNX3 increased in the miR-106b-5p inhibitor group, and the expression levels of miR-106b-5p, MMP2 and MMP9 decreased, and numbers of migrating and invasive cells decreased (P<0.05). Compared with the miR-106b-5p inhibitor+si-RNA group, the miR-106b-5p inhibitor+si-RUNX3 group showed lower RUNX3 expression level, increased miR-106b-5p, MMP2 and MMP9 expression levels, and increased numbers of migrating and invading cells (P<0.05). Conclusion PIC inhibits the miR-106b-5p expression and promotes RUNX3 expression to inhibit CC cell migration and invasion.

Key words: microRNAs, core binding factor alpha 3 subunit, uterine cervical neoplasms, cell movement, neoplasm invasiveness, piceatannol, miRNA-106b-5p, RUNT-related transcription factor 3

CLC Number:

R285，R349.5

Figures/Tables 9

References 19

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组别	miR-106b-5p	RUNX3	迁移数量/（个/视野）	侵袭数量/（个/视野）	MMP2	MMP9
Control组	1.00±0.03	0.31±0.02	296.31±33.24	221.27±20.16	1.24±0.10	1.35±0.13
PIC组	0.24±0.04^a	1.12±0.10^a	94.20±4.67^a	64.38±8.17^a	0.37±0.04^a	0.42±0.04^a
PIC+NC mimics组	0.28±0.05^a	1.15±0.12^a	98.86±4.97^a	69.31±6.19^a	0.41±0.05^a	0.51±0.06^a
PIC+miR-106b-5p mimics组	0.63±0.07^c	0.71±0.08^c	183.31±15.83^c	137.29±11.54^c	0.79±0.08^c	0.92±0.09^c
F	304.869^＊＊	120.789^＊＊	153.635^＊＊	200.789^＊＊	192.263^＊＊	144.477^＊＊

组别	miR-106b-5p	RUNX3	迁移数量/（个/视野）	侵袭数量/（个/视野）	MMP2	MMP9
Control组	1.00±0.03	0.31±0.02	296.31±33.24	221.27±20.16	1.24±0.10	1.35±0.13
PIC组	0.24±0.04^a	1.12±0.10^a	94.20±4.67^a	64.38±8.17^a	0.37±0.04^a	0.42±0.04^a
PIC+NC mimics组	0.28±0.05^a	1.15±0.12^a	98.86±4.97^a	69.31±6.19^a	0.41±0.05^a	0.51±0.06^a
PIC+miR-106b-5p mimics组	0.63±0.07^c	0.71±0.08^c	183.31±15.83^c	137.29±11.54^c	0.79±0.08^c	0.92±0.09^c
F	304.869^＊＊	120.789^＊＊	153.635^＊＊	200.789^＊＊	192.263^＊＊	144.477^＊＊

组别	RUNX3-WT	RUNX3-MUT
miR-NC组	1.00±0.01	1.01±0.02
miR-106b-5p mimics组	0.29±0.05	0.99±0.03
t	34.107^＊＊	1.359

组别	RUNX3-WT	RUNX3-MUT
miR-NC组	1.00±0.01	1.01±0.02
miR-106b-5p mimics组	0.29±0.05	0.99±0.03
t	34.107^＊＊	1.359

组别	miR-106b-5p	RUNX3	迁移数量/（个/视野）	侵袭数量/（个/视野）	MMP2	MMP9
NC inhibitor组	1.02±0.05	0.25±0.03	284.29±30.74	211.28±20.93	1.30±0.11	1.42±0.14
miR-106b-5p inhibitor组	0.32±0.04^a	0.99±0.08^a	86.14±3.20^a	51.27±7.38^a	0.31±0.04^a	0.37±0.03^a
miR-106b-5p inhibitor+si-RNA组	0.28±0.04^a	1.07±0.12^a	93.75±4.18^a	58.16±3.15^a	0.27±0.03^a	0.30±0.05^a
miR-106b-5p inhibitor+si-RUNX3组	0.57±0.06^c	0.63±0.05^c	171.23±13.16^c	143.14±11.65^c	0.85±0.07^c	1.05±0.10^c
F	298.559^＊＊	140.000^＊＊	177.429^＊＊	217.499^＊＊	294.677^＊＊	213.552^＊＊