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沉默NEUROG3基因表达增强胃癌细胞恶性表型的实验研究
Neurogenin 3 silencing promotes the malignant phenotype of gastric cancer cells
- 第三军医大学学报 2020年42卷第10期 页码：1008-1014
- 作者机构：
1.400010 重庆，重庆医科大学附属第二医院：胃肠外科
2.400010 重庆，重庆医科大学附属第二医院：病理科
- 作者简介：
[ 张剑波，E-mail:zhangjianbo@hospital.cqmu.edu.cn ZHANG Jianbo, E-mail:zhangjianbo@hospital.cqmu.edu.cn
- 基金信息：重庆市卫生计生委2017年医学科研计划项目(2017MSXM033);重庆市基础科学与前沿技术研究项目(cstc2017jcyjAX0224)
DOI：10.16016/j.1000-5404.201911237
中图分类号： R394.3;R730.23;R735.2收稿：2019-11-28，
修回：2020-02-10，
纸质出版：2020-05-30
- 稿件说明：
移动端阅览
蒋丹丹, 陈茜, 赵海霖, 等. 沉默NEUROG3基因表达增强胃癌细胞恶性表型的实验研究[J]. 第三军医大学学报, 2020,42(10):1008-1014.
Dandan JIANG, Xi CHEN, Hailin ZHAO, et al. Neurogenin 3 silencing promotes the malignant phenotype of gastric cancer cells[J]. Journal of Third Military Medical University, 2020, 42(10): 1008-1014.
蒋丹丹, 陈茜, 赵海霖, 等. 沉默NEUROG3基因表达增强胃癌细胞恶性表型的实验研究[J]. 第三军医大学学报, 2020,42(10):1008-1014. DOI： 10.16016/j.1000-5404.201911237.
Dandan JIANG, Xi CHEN, Hailin ZHAO, et al. Neurogenin 3 silencing promotes the malignant phenotype of gastric cancer cells[J]. Journal of Third Military Medical University, 2020, 42(10): 1008-1014. DOI： 10.16016/j.1000-5404.201911237.
摘要
目的
2
检测神经元素3(neurogenin3，NEUROG3)在胃癌临床标本和细胞系中的表达，探讨其对胃癌细胞恶性表型的影响。
方法
2
采用免疫组织化学染色(immunohistochemistry，IHC)检测NEUROG3在肠化生胃黏膜和胃癌组织中的表达。利用Real-time PCR和Western blot检测NEUROG3在不同分化程度胃癌细胞系和人胃黏膜上皮细胞系GES-1中的表达。使用慢病毒转染沉默NEUROG3基因表达，Real-time PCR检测胃癌细胞分化抑制蛋白(inhibitor of DNA binding or differentiation，Id-1)的表达，利用EdU细胞增殖实验和细胞克隆形成实验检测胃癌细胞的增殖，划痕实验和Transwell实验检测胃癌细胞迁移和侵袭。
结果
2
NEUROG3在低分化胃癌组织中表达较低，而在癌旁组织、肠化生胃黏膜组织和中高分化胃癌组织中表达评分较高，评分差异有统计学意义(
P
<
0.001)。NEUROG3的表达差异与患者性别、年龄和肿瘤大小无统计学意义(
P
>
0.05)，而与肿瘤分化程度(
P
<
0.001，
r
=0.412)呈正相关，与淋巴结转移(
P
<
0.05，
r
=-0.180)和TNM分期呈负相关(
P
<
0.001，
r
=-0.431)。GES-1和高分化胃癌细胞系MKN-28中
的NEUROG3表达明显高于中低分化胃癌细胞系SGC-7901、MKN-45和BGC-823(
P
<
0.001)。MKN-28细胞沉默NEUROG3表达后，细胞分化程度降低，Id-1表达明显增高(
P
<
0.001)，细胞增殖、迁移和侵袭能力(
P
<
0.01)明显增强。
结论
2
沉默NEUROG3基因表达，可抑制胃癌细胞的分化，促进胃癌细胞增殖、迁移和侵袭。
Abstract
Objective
2
To detect the expression of neurogenin 3 in gastric cancer tissues and cultured gastric cancer cells and determine its role in regulating the malignant phenotype of gastric cancer cells.
Methods
2
Immunohistochemical staining was used to detect the expression of neurogenin 3 in tissue specimens of intestinal metaplasia gastric mucosa and gastric cancer. The expression of neurogenin 3 in human gastric mucosal epithelial cells (GES-1) and in gastric cancer cell lines with different levels of differentiation was detected using real-time PCR and Western blotting. In gastric cancer MKN-28 cells
the effect of neurogenin 3 gene silencing mediated by lentivirus transfection on the expression of cell differentiation inhibitor of DNA binding or differentiation (Id-1) was detected using real-time PCR. The changes in the proliferation of the cells were assessed using EdU cell proliferation assay and cell colony formation assay
and the migration and invasion of the transfected cells were evaluated with a scratch assay and Transwell assay.
Results
2
Neurogenin 3 was lowly expressed in poorly differentiated gastric cancer tissues but highly expressed in adjacent tissues
intestinal metaplasia gastric mucosa tissues and moderately to well differentiated gastric cancer tissues (
P
<
0.001). Neurogenin 3 expression in gastric cancer tissue showed no significant correlation with the patients' gender
age or tumor size (
P
>
0.05)
but was significantly correlated with the level of tumor differentiation (
P
<
0.001
r
=0.412)
lymph node metastasis (
P
<
0.05
r
=-0.180) and TNM staging (
P
<
0.001
r
=-0.431). The expression of neurogenin 3 in well differentiated gastric cancer MKN-28 and gastric GES-1 cell lines was significantly higher than that in moderately or poorly differentiated gastric cancer SGC-7901
MKN-45 and BGC-823 cell lines. Neurogenin 3 silencing in MKN-28 cells resulted in a lower level of cell differentiation
a significantly increased expression of Id-1 (
P
<
0.001) and significantly enhanced cell proliferation
migration and invasion (
P
<
0.01).
Conclusion
2
Neurogenin 3 silencing can inhibit the differentiation and promote the proliferation
migration and invasion of gastric cancer cells
in vitro
.
关键词
Keywords
references
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