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1.重庆市渝北区人民医院消化内科,重庆401120
2.陆军军医大学第一附属医院麻醉科,重庆 400038
田音,E-mail:tianyindoctor@163.com
收稿:2024-11-15,
纸质出版:2025-12-25
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杨清清,曾宇,田音.N-糖基化调控NANOG在结直肠癌干细胞特性和凋亡中的影响[J].局解手术学杂志,2025,34(12):1063-1068.
YANG Qing-qing,ZENG Yu,TIAN Yin.N-glycosylation regulates NANOG affecting stem cell properties and apoptosis of colorectal cancer[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2025,34(12):1063-1068.
杨清清,曾宇,田音.N-糖基化调控NANOG在结直肠癌干细胞特性和凋亡中的影响[J].局解手术学杂志,2025,34(12):1063-1068. DOI: 10.11659/jjssx.11E024083.
YANG Qing-qing,ZENG Yu,TIAN Yin.N-glycosylation regulates NANOG affecting stem cell properties and apoptosis of colorectal cancer[J].JOURNAL OF REGIONAL ANATOMY AND OPERATIVE SURGERY,2025,34(12):1063-1068. DOI: 10.11659/jjssx.11E024083.
目的
2
探究N-糖基化对NANOG调控结直肠癌干细胞特性的影响。
方法
2
对结直肠癌HCT166细胞和LoVo细胞进行磁珠分选筛选出CD133
+
干细胞。采用Lipofectamine™ 3000对结直肠癌干细胞进行质粒转染。MTT法检测细胞活性。肿瘤干细胞成球实验检测干细胞成球直径和成球数量。EdU流式细胞术检测细胞增殖。划痕实验检测细胞迁移能力。Western blot检测细胞凋亡相关蛋白表达水平。
结果
2
与WT组相比,MUT组经N-糖基化位点缺失的突变表达载体转染的结直肠癌干细胞细胞活性降低(
P
<
0.05),增殖和迁移能力减弱(
P
<
0.05),肿瘤干细胞成球能力降低(
P
<
0.05),细胞内凋亡水平上升(
P
<
0.05)。
结论
2
人源NANOG羧基端内7个N-糖苷型糖链参与NANOG蛋白的分子质量控制及结直肠癌干细胞的干细胞特性维持,进一步影响结直肠癌干细胞的增殖迁移能力。
Objective
2
To explore the effect of N-glycosylation on the properties of colorectal cancer stem cells regulated by NANOG.
Methods
2
CD133
+
stem cells were screened out by magnetic bead sorting on colorectal cancer HCT166 cells and LoVo cells. Plasmid transfection of colorectal cancer stem cells was performed by Lipofectamine™ 3000.MTT assay was used to detect cell activity. Sphere formation assay was used to detect stem cell spheroid diameter and spheroid number.EdU flow cytometry was used to detect cell proliferation. Scratch wound healing assay was used to detect cell migration ability. Western blot was used to detect the expression levels of apoptosis-related proteins.
Results
2
Compared with the WT group,the activity of colorectal cancer stem cells transfected with a mutant expression ve
ctor lacking N-glycosylation sites was reduced(
P
<
0.05),the proliferation and migration abilities were weakened(
P
<
0.05),the spherogenic ability of cancer stem cells was reduced(
P
<
0.05),and the level of intracellular apoptosis was increased in MUT group(
P
<
0.05).
Conclusion
2
The seven N-glycosidic sugar chains in the carboxyl terminus of human NANOG are involved in the molecular quality control of NANOG protein and the maintenance of stem cell characteristics of colorectal cancer stem cells,further affecting the proliferation and migration ability of colorectal cancer stem cells.
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