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

Tab.1

Fig. 1

Tab.2

Tab.3

Fig.2

Fig.3

Tab.4

Tab.5

Fig. 4

Fig. 5

References 20

1	BRAY F， FERLAY J， SOERJOMATARAM I， et al. Global cancer statistics 2018： GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries［J］. CA Cancer J Clin，2018，68（6）：394-424.
2	曹雅杰，包晓红，李书珍，等. 胰岛素样生长因子1受体抑制剂NVP-AEW541对ESCC细胞增殖、迁移和侵袭的影响及其机制［J］. 吉林大学学报（医学版）， 2021， 47（5）： 1131-1138.
3	CUI J， WEN Q Q， TAN X J， et al. A genomic-clinicopathologic nomogram predicts survival for patients with laryngeal squamous cell carcinoma［J］. Dis Markers， 2019， 2019： 5980567.
4	张璐瑶，汪丽钰，陈乾，等. m6A甲基化修饰与消化道恶性肿瘤关系的研究进展［J］. 医学综述， 2021， 27（9）： 1753-1758.
5	LI F， ZHAO D， WU J， et al. Structure of the YTH domain of human YTHDF2 in complex with an m6A mononucleotide reveals an aromatic cage for m6A recognition［J］. Cell Res，2014，24（12）：1490-1492.
6	牛云峰，杨阳，王欣晨，等. 食管癌组织中PTPN12的表达及其对Yes-2细胞恶性生物学行为的影响［J］. 临床与实验病理学杂志， 2021， 37（6）： 643-649.
7	ZHU J， MA S R， CHEN R， et al. Biological correlates before esophageal cancer screening and after diagnosis［J］. Sci Rep， 2021， 11（1）： 17015.
8	刘翼，祝琳，康敏. miR-302b-3p对食管癌EC-109细胞增殖和凋亡的影响及其机制［J］. 吉林大学学报（医学版）， 2019， 45（6）： 1346-1352.
9	FANG R P， CHEN X， ZHANG S C， et al. EGFR/SRC/ERK-stabilized YTHDF2 promotes cholesterol dysregulation and invasive growth of glioblastoma［J］. Nat Commun， 2021， 12（1）： 177.
10	王月帆，葛春梅，尹昊瓒，等. m6A甲基化修饰识别蛋白YTHDF2在肝癌组织中的表达及临床意义［J］. 现代生物医学进展， 2021， 21（9）： 1601-1606.
11	刘冰，汪涛，臧丽丽，等. RNA甲基化阅读器蛋白YTHDF2对卵巢癌细胞增殖的调控作用［J］. 解剖科学进展， 2021， 27（1）： 68-70， 74.
12	李子玮，罗清雅，王浩丞，等. 敲低YTH结构域N6甲基腺嘌呤（m6A）RNA结合蛋白2（YTHDF2）抑制宫颈癌细胞增殖并促进其凋亡［J］. 细胞与分子免疫学杂志，2020，36（3）：255-263.
13	赵琪，何长海，王志，等. 敲低ALKBH3对膀胱癌细胞生长、迁移与肿瘤血管生成的抑制作用及其机制［J］.吉林大学学报（医学版），2021，47（2）：397-406.
14	LI J F， XIE H Y， YING Y F， et al. YTHDF2 mediates the mRNA degradation of the tumor suppressors to induce AKT phosphorylation in N6-methyladenosine-dependent way in prostate cancer［J］. Mol Cancer， 2020， 19（1）： 152.
15	母润红，刘馨竹，林睿，等. PRDX6过表达对肝癌细胞增殖、侵袭和迁移的影响及其分子机制［J］. 吉林大学学报（医学版）， 2021， 47（3）： 559-565.
16	许瀚，孟祥瑞，周悦，等.FNDC1蛋白在食管癌组织中的表达及干扰FNDC1表达对食管癌细胞生物学行为的影响［J］. 郑州大学学报（医学版），2020，55（4）： 445-450.
17	XIE H Y， LI J F， YING Y F， et al. METTL3/YTHDF2 m6A axis promotes tumorigenesis by degrading SETD7 and KLF4 mRNAs in bladder cancer［J］. J Cell Mol Med， 2020， 24（7）： 4092-4104.
18	HUANG T， LIU Z D， ZHENG Y， et al. YTHDF2 promotes spermagonial adhesion through modulating MMPs decay via m6A/mRNA pathway［J］. Cell Death Dis， 2020， 11（1）： 37.
19	LI Y， SHENG H， MA F， et al. RNA m6A reader YTHDF2 facilitates lung adenocarcinoma cell proliferation and metastasis by targeting the AXIN1/Wnt/β-catenin signaling［J］.Cell Death Dis，2021，12（5）： 479.
20	ZHANG C Z， HUANG S Z， ZHUANG H K， et al. YTHDF2 promotes the liver cancer stem cell phenotype and cancer metastasis by regulating OCT4 expression via m6A RNA methylation［J］. Oncogene， 2020， 39（23）： 4507-4518.

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^*

[1]	Zhijuan WANG,Mingshu ZHANG,Liping YE. Effects of platelet-derived growth factor D on proliferation, migration and invasion of lung cancer H1299 cells through ERK signaling pathway and their mechanisms [J]. Journal of Jilin University(Medicine Edition), 2022, 48(4): 898-904.
[2]	Weibo LI,Juan CAO,Xiu GUO,Chunling DONG,Bo LI. Promotion effect of tumor associated macrophages-derived IL-6 on invasion and migration of oral squamous cell carcinoma Cal27 cells by upregulating LIF expression in tumor cells [J]. Journal of Jilin University(Medicine Edition), 2022, 48(4): 946-953.
[3]	Guowu WANG,Yuan YAO,Yu ZHANG,Na XU,Fang LIU. Inhibitory effect of miR-152 on proliferation and invasion of endometrial carcinoma cells by reducing low-density lipoprotein receptor expression [J]. Journal of Jilin University(Medicine Edition), 2022, 48(3): 591-599.
[4]	Guanhu LI,Qingxu LANG,Chunyan LIU,Qin LIU,Mengrou GENG,Xiaoqian LI,Zhenqi WANG. Inhibitory effect of valproic acid combined with X-ray irradiation on proliferation of breast cancer MDA-MB-231 cells and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2022, 48(3): 622-629.
[5]	Peiyu YAN,Aichen ZHANG,Hong ZHANG,Yang LI,Mengmeng ZHANG,Mengze LUO,Ying PAN. Therapeutic effect of adipose-derived mesenchymal stem cells on premature ovarian failure model rats and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2022, 48(3): 648-656.
[6]	Cuilan LIU,Fengai HU,Jing LIU,Dan WANG,Changyun QIU,Dunjiang LIU,Di ZHAO. Effect of adiponectin receptor agonist AdiopRon on biological behaviors of glioma cells and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2022, 48(3): 702-710.
[7]	Zhuangzhi WU,Xiaoning HE,Siqi CHEN. Inhibitory effect of miR-124-3p on proliferation and invasion of oral squamous cell carcinoma cells and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2022, 48(3): 718-727.
[8]	Xiaoyan LI,Wei ZHANG,Jie HE. Promotion effect of REG1A on proliferation and migration of lung adenocarcinoma cells by regulating Wnt/β-catenin signaling pathway [J]. Journal of Jilin University(Medicine Edition), 2022, 48(2): 444-453.
[9]	Guangsong XU,Haibing JIANG,Jing PAN,Guoqing LI. Inhibitory effects of betulinic acid on migration and invasion of gastric cancer MGC-803 cells and their mechanisms [J]. Journal of Jilin University(Medicine Edition), 2022, 48(1): 122-128.
[10]	Chaofeng ZHOU,Shifan ZHOU,Qing TIAN,Sai WANG,Honglin LI,Chunzheng MA. Effect of lncRNA-NORAD overexpression on biological behaviors of esophageal cancer Eca-109 cells and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2022, 48(1): 33-43.
[11]	Zetai WANG,Dandan LOU,Yan PENG,Daoqi ZHU,Aiwu LI,Fengying GONG,Ying LYU,Qin FAN. Establishment of zebrafish xenograft model of nasopharyngeal carcinoma and inhibitory effect of curcumin on CNE-2 cells [J]. Journal of Jilin University(Medicine Edition), 2022, 48(1): 9-17.
[12]	Runhong MU,Yijiu AI,Yupeng LI,Rui LIN,Siping YE,Fang MA,Xiao GUO. Expression of recombinant human IL-17A in gastric cancer tissue and its effects on proliferation, invasion, migration and apoptosis of gastric cancer BGC-823 cells [J]. Journal of Jilin University(Medicine Edition), 2021, 47(6): 1510-1517.
[13]	Naigao TANG,Genjian ZHENG. Effects of miR-762 on proliferation and apoptosis of human tongue squamous cell carcinoma cells by targeting NCOR1 expression [J]. Journal of Jilin University(Medicine Edition), 2021, 47(5): 1099-1107.
[14]	Yajie CAO,Xiaohong BAO,Shuzhen LI,Haiying GENG,Zengxiaorui CAI,Chunmei DAI,Ning LI. Effects of insulin-like growth factor 1 receptor inhibitor NVP-AEW541 on proliferation, migration and invasion of ESCC cells [J]. Journal of Jilin University(Medicine Edition), 2021, 47(5): 1131-1138.
[15]	Chen WANG,Jun TIAN,Hailing CHENG. Effect of human umbilical cord mesenchymal stem cells on proliferation and apoptosis of cervical cancer HeLa cells and its mechanism [J]. Journal of Jilin University(Medicine Edition), 2021, 47(5): 1187-1193.

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

RichHTML

PDF (PC)

Like

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 10

References 20

Related Articles 15

Metrics

Comments

Recommended 0