Effect of miR-491-5p's regulation of SHOC2 on the proliferation, migration and invasion of esophageal squamous cell carcinoma cells

doi:10.3877/cma.j.issn.2095-1221.2019.06.003

Abstract

Abstract:

Objective

To investigate the effect of miR-491-5p on proliferation, migration and invasion of esophageal squamous cell carcinoma (ESCC) cells and its mechanism.

Methods

The immortalized esophageal epithelial cell line HET-1A and human esophageal cancer cell lines EC109, EC9706, KYSE510 were cultured, and the levels of miR-491-5p and SHOC2 mRNA were detected by qRT- PCR. EC109 cells were divided into control group, miR-491-5p group, miR-NC group, miR-491-5p+pcDNA-SHOC2 group and miR-491-5p+pcDNA group. MTT was used to detect cell proliferation, Transwell was used to detect cell migration and invasion, and Western Blot was used to detect the expression levels of CyclinD1, Vimentin, E-cadherin and MAPK/ERK signaling pathway-related protein. The dual luciferase reporter gene assay was used to verify the regulatory relationship between miR-491-5p and SHOC2. The comparison between the two groups was performed using independent sample t tests. One-way analysis of variance was used for comparison between multiple groups, and SNK-q test was used for further comparisons.

Results

The expression levels of miR- 491-5p in esophageal squamous cells EC109, EC9706 and KYSE510 were lower than those in HET-1A cells (0.32±0.06, 0.62±0.10, 0.61±0.08 vs 1.00±0.08, F = 106.340, P < 0.001) ; The mRNA expression level of SHOC2 was higher than that of HET-1A cells (2.85±0.16, 1.73±0.10, 1.45±0.06 vs 1.02±0.09, F = 464.949, P < 0.001) . The OD value, cell migration number, invasion number, and CyclinD1, Vimentin, p-MEK, and p-ERK protein levels in miR-491- 5p group of EC109 cells after 72 h of culture were lower than those in miR-NC group (0.70±0.06 vs 1.42±0.08, 65.01±10.36 vs 150.01±12.48, 70.03±10.26 vs 140.02±11.85, 0.30±0.03 vs 0.93±0.16, 0.41±0.05 vs 0.86±0.08, 0.32±0.06 vs 0.95±0.11, 0.40±0.06 vs 0.92±0.13, F = 236.565, 159.440, 120.706, 101.071, 98.619, 130.766, 77.046, P < 0.001) , E-cadherin protein levels were higher than miR-NC group (0.89±0.13 vs 0.48±0.08, F = 816.432, P < 0.001) . miR-491-5p negatively regulated SHOC2 expression in EC109 cells. SHOC2 overexpression reversed the effect of miR-491-5p overexpression on proliferation, migration and invasion of EC109 cells and MAPK/ERK signaling pathway.

Conclusion

miR-491-5p can inhibit the proliferation, migration and invasion of ESCC cells, and its mechanism may be related to the down-regulation of SHOC2 expression and inhibition of MAPK/ERK signaling pathway activity.

Key words: Esophageal squamous cell carcinoma, miR-491-5p, SHOC2, Cell proliferation, Cell migration, Cell invasion

Ye Xu. Effect of miR-491-5p's regulation of SHOC2 on the proliferation, migration and invasion of esophageal squamous cell carcinoma cells[J]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2019, 09(06): 333-339.

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Figures/Tables 13

References 17

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分组	细胞活性（OD 490 nm）			迁移细胞数	侵袭细胞数	CyclinD1	Vimentin	E-cadherin	miR-491-5p
分组	24 h	48 h	72 h	迁移细胞数	侵袭细胞数	CyclinD1	Vimentin	E-cadherin	miR-491-5p
空白对照组	0.43±0.06	0.86±0.11	1.43±0.10	156±13.25	145±12.15	0.95±0.10	0.87±0.10	0.46±0.11	1.00±0.03
miR-NC组	0.41±0.03	0.89±0.13	1.42±0.08	150±12.48	140±11.85	0.93±0.16	0.86±0.08	0.48±0.08	1.01±0.05
miR-491-5p组	0.20±0.02^a	0.52±0.10^a	0.70±0.06^a	65±10.36^a	70±10.26^a	0.30±0.03^a	0.41±0.05^a	0.89±0.13^a	3.45±0.25
F值	89.449	29.238	236.565	159.440	120.706	101.071	98.619	44.924	816.432
P值	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001

分组	细胞活性（OD 490 nm）			迁移细胞数	侵袭细胞数	CyclinD1	Vimentin	E-cadherin	miR-491-5p
分组	24 h	48 h	72 h	迁移细胞数	侵袭细胞数	CyclinD1	Vimentin	E-cadherin	miR-491-5p
空白对照组	0.43±0.06	0.86±0.11	1.43±0.10	156±13.25	145±12.15	0.95±0.10	0.87±0.10	0.46±0.11	1.00±0.03
miR-NC组	0.41±0.03	0.89±0.13	1.42±0.08	150±12.48	140±11.85	0.93±0.16	0.86±0.08	0.48±0.08	1.01±0.05
miR-491-5p组	0.20±0.02^a	0.52±0.10^a	0.70±0.06^a	65±10.36^a	70±10.26^a	0.30±0.03^a	0.41±0.05^a	0.89±0.13^a	3.45±0.25
F值	89.449	29.238	236.565	159.440	120.706	101.071	98.619	44.924	816.432
P值	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001	< 0.001

分组	SHOC2-WT	SHOC2-MUT
miR-NC组	1.06±0.05	1.05±0.10
miR-491-5p组	0.45±0.02^a	1.02±0.06
t值	33.982	0.772
P值	< 0.001	0.452

分组	SHOC2-WT	SHOC2-MUT
miR-NC组	1.06±0.05	1.05±0.10
miR-491-5p组	0.45±0.02^a	1.02±0.06
t值	33.982	0.772
P值	< 0.001	0.452

分组	SHOC2蛋白
miR-NC组	0.80±0.06
miR-491-5p组	0.25±0.02^a
anti-miR-NC组	0.73±0.09
anti-miR-491-5p组	0.98±0.10^b
F值	158.226
P值	< 0.001

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