circRNA_0005075在胃癌中的表达及对胃癌细胞增殖、侵袭能力的调控机制

窦广健; 黄立勇; 吴佳明; 李进; 陈治横

doi:10.16766/j.cnki.issn.1674-4152.002928

circRNA_0005075在胃癌中的表达及对胃癌细胞增殖、侵袭能力的调控机制

doi: 10.16766/j.cnki.issn.1674-4152.002928

嘉兴市第一医院(嘉兴学院附属医院)胃肠外科，浙江嘉兴 314000

基金项目:

浙江省医药卫生科技计划项目 2020RC121

浙江省嘉兴市科技计划项目 2019AD32262

详细信息

通讯作者:
黄立勇，E-mail：moegq0@163.com

中图分类号: R735.2 R730.231
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- 文章访问数: 146
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- 被引次数: 0
出版历程
- 收稿日期: 2022-12-02
- 网络出版日期: 2023-05-31

Expression of circRNA_0005075 in gastric cancer and the regulation mechanism of cell proliferation and invasion of gastric cancer cells

Department of Gastrointestinal Surgery, Jiaxing First Hospital (Affiliated Hospital of Jiaxing University), Jiaxing, Zhejiang 314000, China

摘要

摘要: 目的环状RNA (circRNA)与胃癌发生发展有关, 本研究旨在探讨circRNA_0005075对胃癌细胞增殖、侵袭的影响及其对miR-767-5p/TRIM27轴的调控作用。方法采用RT-qPCR和RNA FISH法检测circRNA_0005075在胃癌组织和配对癌旁组织中的表达水平。用慢病毒感染技术过表达胃癌细胞的circRNA_0005075, 并通过体内RNA免疫共沉淀技术、Western blotting、荧光素酶活性实验和挽救实验对其分子作用机制进行探讨。结果癌组织circRNA_0005075相对表达量为0.51±0.13, 低于癌旁组织的3.51±0.69(P < 0.05)。癌旁组织中circRNA_0005075的荧光强度OD值为1.35±0.24, 高于癌组织的0.67±0.16(P < 0.05)。circRNA_0005075低表达的胃癌患者总体生存时间显著短于高表达患者(P < 0.05)。circRNA_0005075过表达可抑制胃癌细胞的增殖和侵袭。RT-qPCR及荧光素酶活性实验证实circRNA_0005075可与miR-767-5p结合。利用信息预测网站(http://www.targetscan.org)预测到miR-767-5p与TRIM27结合位点。Western blotting实验证实, circRNA_0005075可通过调控miR-767-5p表达进而抑制TRIM27的表达。结论 circRNA_0005075通过靶向作用于miR-767-5p/TRIM27轴抑制胃癌细胞的增殖和侵袭。
- 胃癌 /
- 环状RNA /
- micro RNA /
- 细胞增殖 /
- 细胞侵袭
Abstract: Objective Circular RNA (circRNA) is related with gastric carcinogenesis and development.The aim of this study was to investigate the effect of circRNA_0005075 on gastric cancer cell proliferation and invasion and its regulation of the miR-767-5p/TRIM27 axis. Methods The expression level of circRNA_0005075 in gastric cancer tissues and paired adjacent tissues was determined by RT-qPCR and RNA FISH.Lentiviral infection technique was used to overexpress circRNA_0005075 in gastric cancer cells, and the molecular mechanism was explored by in vivo RNA immunoprecipitation, western blotting, luciferase activity assay and rescue experiments. Results The relative expression of circRNA_0005075 was 0.51±0.13, which was lower than that in adjacent tissues (3.51±0.69, P < 0.05).The fluorescence intensity OD value of circRNA_0005075 in paracancerous tissue was 1.35±0.24, which was higher than that in cancerous tissue (0.67±0.16, P < 0.05).The overall survival time of gastric cancer patients with low expression of circRNA_0005075 was significantly shorter than that of patients with high expression (P < 0.05).circRNA_0005075 overexpression can inhibit the proliferation and invasion of gastric cancer cells.RT-qPCR and luciferase activity test confirmed that circRNA_0005075 can be combined with miR-767-5p.The binding site of miR-767-5p with TRIM27 was predicted by using the information website (http://www.targetscan.org).Western blotting experiments confirmed that circRNA_0005075 could inhibit the expression of TRIM27 by regulating the expression of miR-767-5p. Conclusion circRNA_0005075 inhibits the proliferation and invasion of gastric cancer cells by targeting the miR-767-5p/TRIM27 axis.
- Gastric cancer /
- CircRNA /
- Micro RNA /
- Cell proliferation /
- Cell invasion

HTML全文

图 1 不同circRNA_0005075表达水平患者的生存曲线

Figure 1. Survival curves of patients with different expression levels of circRNA_0005075

下载: 全尺寸图片幻灯片

图 2 胃癌细胞中circRNA_0005075与miR-767-5p结合的验证

注：A示过表达circRNA_0005075胃癌细胞系的验证；B示过表达circRNA_0005075对miRNA表达水平的影响；C示RIP结合RT-qPCR实验验证ircRNA_0005075与miR-767-5p结合。

Figure 2. Validation of circRNA_0005075 binding to miR-767-5p in gastric cancer cells

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图 3 circRNA_0005075过表达对胃癌细胞增殖能力的影响

注：A示CCK-8实验检测细胞增殖能力；B示EDU实验检测细胞增殖能力。

Figure 3. Effect of circRNA_0005075 overexpression on proliferation of gastric cancer cells

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图 4 circRNA_0005075对miR-767-5p/TRIM27通路的调控作用

注：A示miR-767-5p和TRIM27的结合位点；B示荧光素酶报告基因实验证实miR-767-5p对TRIM27的调控；C示circRNA_0005075通过调控miR-767-5p对TRIM27蛋白表达的影响。

Figure 4. Regulation of miR-767-5p/TRIM27 pathway by circRNA_0005075

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表 1 circRNA_0005075与临床病理参数的关系(例)

Table 1. Relationship between circRNA _0005075 and clinicopathological parameters(cases)

因素	例数	circRNA_0005075		χ²值	P值
因素	例数	低表达(n=50)	高表达(n=50)	χ²值	P值
年龄(岁)				0.208	0.648
≥60	74	38	36
＜60	26	12	14
性别(例)				0.184	0.668
男性	68	35	33
女性	32	15	17
肿瘤大小(cm)				5.797	0.016
≥5	46	29	17
＜5	44	21	33
Lauren分型(例)				0.041	0.839
肠型	59	29	30
弥漫型	41	21	20
T分期(例)				5.086	0.024
T1+T2	39	14	25
T3 +T4	61	36	25
淋巴结转移				8.167	0.004
有	60	37	23
无	40	13	27
M分期(例)				6.832	0.009
M0	77	33	44
M1	23	17	6
脉管浸润(例)				6.618	0.010
有	32	22	10
无	68	28	40

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表 2 影响胃癌患者生存的Cox多因素分析

Table 2. Cox multivariate analysis of factors affecting the survival of gastric cancer patients

变量	β	SE	Waldχ²	P值	HR	95% CI
年龄	0.867	0.325	7.138	0.008	2.553	1.262~4.503
性别	-0.594	0.352	2.935	0.084	0.548	0.274~1.092
肿瘤大小	1.276	0.588	4.746	0.029	3.575	1.022~11.910
T分期	0.262	0.264	1.117	0.040	1.311	0.768~1.894
N分期	0.478	0.632	5.521	0.022	2.125	0.956~3.202
M分期	0.325	0.158	16.521	< 0.001	3.584	2.168~9.639
circRNA_0005075表达	-1.573	0.187	77.633	0.003	0.330	0.103~0.598
脉管浸润	-0.451	0.283	2.592	0.032	0.631	0.362~1.125
淋巴结转移	1.202	0.184	22.001	< 0.001	3.782	1.098~7.231

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