m6A甲基化修饰结合蛋白YTHDF2在食管癌组织中的表达及其对食管癌细胞增殖和迁移的影响

doi:10.13481/j.1671-587X.20220416

Abstract

Abstract:

Objective To investigate the expression of N6-methyladenosine（m6A） methylation binding protein YTH domain-containing family protein 2 （YTHDF2） in the esophageal squamous cell carcinoma （ESCC） tissue and its effect on the biological behavior of esophageal carcinoma cells，and to clarify its possible mechanism.

Methods

The expressions of YTHDF2 protein in 113 cases of ESCC tissue and 95 cases of normal tissue adjacent to esophageal carcinoma were detected by immunohistochemical method. The relationships between the expression of YTHDF2 protein and the clinicopathological features and prognosis of the ESCC patients were analyzed.The Eca109 and EC9706 cells cultured in vitro were divided into control group （transfected with si-NC） and si-YTHDF2 groups （transfected with si-YTHDF2#1 and si-YTHDF2#2）. The expressions of YTHDF2 protein in the cells in various groups were detected by Western blotting method. The proliferation abilities of cells in various groups were detected by CCK-8 method， the number of colony formation in various groups was detected by colony formation experiment， and the number of migration cells in various groups were detected by Transwell chamber assay.

Results

The YTHDF2 protein mainly expressed in the cytoplasm of ESCC cells and a small amount of YTHDF2 protein expressed in the nucleus.The expression amount of YTHDF2 protein in ESCC tissue was significantly higher than that in paracancerous normal tissue （P<0.01）. There were significant differences in the expression intensities of YTHDF2 protein in the ESCC patients with different ages of onset and TNM stages （P=0.008，P=0.041）. The Kaplan-Meier survival analysis showed that compared with YTHDF2 low expression group，the survival time of the patients in YTHDF2 high expression group was significantly shortened（P<0.05）. The CCK-8 method and colony formation experiment results showed that the proliferation activities and the number of colony formation of esophageal carcinoma cells in si-YTHDF2 transfection groups were significantly lower than those in control group （P<0.01）. The Transwell chamber assay results showed that the number of migration esophageal carcinoma cells in si-YTHDF2 transfection groups was significantly lower than that in control group （P<0.01）.

Conclusion

The expression of YTHDF2 in ESCC tissue is significantly higher than that in the normal tissue. YTHDF2 knockdown contributes to inhibiting the proliferation， colony formation and migration of esophageal carcinoma cells.

Key words: Esophageal neoplasms, Squamous cell carcinoma, YTH domain-containing family protein 2, Cell proliferation, Cell migration

CLC Number:

R735.1

Yandi MA,Xiangyun LU,Shangfeng HE,Xueyan YU,Yunhua HU,Haixia GAO,Yunzhao CHEN,Jie YU,Wenjie WANG,Feng LI,Xiaobin CUI. Expression of m6A methylation binding protein YTHDF2 in esophageal carcinoma tissue and its effect on proliferation and migration of esophageal carcinoma cells[J].Journal of Jilin University(Medicine Edition), 2022, 48(4): 962-970.

Figures/Tables 10

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References 20

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Group	n	YTHDF2			χ²	P
Group	n	No expression	Low expression	High expression	χ²	P
ESCC tissue	113	0	33	80	180.66	<0.01
Normal tissue	95	58	37	0	180.66	<0.01

Clinicopathological feature	n	YTHDF2		χ²	P
Clinicopathological feature	n	Low expression	High expression	χ²	P
Gender
Male	38	13	25	0.432	0.580
Female	21	9	12	0.432	0.580
Age of onset（year）
<65	41	20	21	7.590	0.008
≥65	18	2	16	7.590	0.008
Location
Up & Middle	42	14	28	0.975	0.380
Low	17	8	9	0.975	0.380
Grade
G1	19	7	12	1.148	0.563
G2	28	12	16
G3	12	3	9
Tumor
T1+T2	20	7	13	0.068	1.000
T3	39	15	24	0.068	1.000
Node
N0	26	11	15	0.501	0.590
N1	33	11	22	0.501	0.590
Metastatis
M0	52	21	31	1.797	0.240
M1	7	1	6	1.797	0.240
TNM
Ⅰ+Ⅱ+Ⅲ	48	21	27	4.597	0.041
Ⅳ	11	1	10	4.597	0.041

Group	Proliferation activity
Group	(t/d) 1	2	3	4	5
Control	0.205±0.002	0.298±0.004	0.422±0.007	0.626±0.002	1.392±0.004
si-YTHDF2#1	0.200±0.005	0.277±0.001^*	0.347±0.007^*	0.442±0.013^*	0.883±0.022^*
si-YTHDF2#2	0.214±0.006	0.305±0.005	0.349±0.006^*	0.378±0.004^*	0.548±0.009^*

Group	Proliferation activity
Group	(t/d) 1	2	3	4	5
Control	0.264±0.004	0.306±0.006	0.441±0.009	0.799±0.025	1.851±0.032
si-YTHDF2#1	0.252±0.002^*	0.281±0.005^*	0.434±0.003	0.520±0.008^*	1.128±0.003^*
si-YTHDF2#2	0.216±0.002^*	0.248±0.003^*	0.365±0.004^*	0.423±0.006^*	0.694±0.022^*

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Expression of m6A methylation binding protein YTHDF2 in esophageal carcinoma tissue and its effect on proliferation and migration of esophageal carcinoma cells

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