Notch1P1641S突变通过PI3K/Akt通路促进口腔鳞癌增殖

doi:10.13591/j.cnki.kqyx.2023.05.002

摘要/Abstract

摘要：

目的探究Notch1^P1641S突变对口腔鳞状细胞癌的影响。方法体外培养口腔鳞癌细胞系CAL27与HSC3,构建Notch1^N1(N1-N1)、Notch1^WT(N1-WT)及Notch1^P1641S(N1-P1641S)位点突变质粒,将质粒转染细胞系中。通过qPCR及Westen blot实验检测细胞系的转染效率。表型实验通过CCK-8、集落形成实验、划痕实验、EdU、Transwell实验检测P1641S位点突变后对口腔鳞癌细胞系的增殖、迁移、侵袭的影响。通过Westen blot检测转染突变质粒后对细胞通路的影响。通过裸鼠皮下成瘤实验进一步验证突变质粒对口腔鳞癌的影响。结果表型实验结果显示转染突变质粒后,相较于N1-N1组及N1-WT组,N1-P1641S组口腔鳞癌细胞系的增殖、迁移、侵袭能力均增强。Westen blot实验结果显示转染突变质粒后,N1-P1641S组PI3K/Akt相关指标表达上调。裸鼠皮下成瘤实验显示转染突变质粒后,裸鼠成瘤体积增大,增殖相关指标Ki67表达增强。结论 Notch1^P1641S表达位点突变通过PI3K/Akt信号通路促进口腔鳞癌细胞增殖。

关键词: 口腔鳞状细胞癌, Notch1, 突变, PI3K

Abstract:

Objective To investigate the effect of Notch1^P1641S mutation on oral squamous cell carcinoma. Methods CAL27 and HSC3 were cultured in vitro. Notch1^N1 (N1-N1), Notch1^WT (N1-WT) and Notch1^P1641S (N1-P1641S) locus mutant plasmids were constructed. Plasmids were transfected into cell lines and the transfection efficiency was examined by qPCR and Westen blot assay. The effects of Notch1 mutation on proliferation, invasion and migration of CAL27 and HSC3 cells were investigated by CCK-8 assay, plate colony formation assay, Scratch assay, EdU assay and Transwell invasion assay. Western blot was used to detect the expression of cellular pathways. The effect of mutant plasmids on oral squamous carcinoma was further verified by subcutaneous tumorigenesis assay in nude mice. Results The results of the phenotypic assay showed that the proliferation, migration and invasion ability of oral squamous carcinoma cell lines in the N1-P1641S group were enhanced after transfection with the mutant plasmid compared to the N1-N1 and N1-WT groups. The results of Westen blot experiments showed that the expression of PI3K/Akt-related indicators was upregulated in the N1-P1641S group after transfection with the mutant plasmid. Subcutaneous tumorigenesis in nude mice showed an increase in tumor size after transfection with the mutant plasmid. The expression of Ki67, a proliferation-related indicator, was enhanced. Conclusion Notch1^P1641S expression site mutation promotes proliferation of oral squamous carcinoma cells through PI3K/Akt signaling pathway.

Key words: oral squamous cell carcinoma, Notch1, mutation, PI3K

中图分类号:

R739.85

杨月美,宋晓萌,吴煜农. Notch1^P1641S突变通过PI3K/Akt通路促进口腔鳞癌增殖[J]. 口腔医学, 2023, 43(5): 393-399.

YANG Yuemei,SONG Xiaomeng,WU Yunong. Promotion of cell proliferation by Notch1^P1641S mutation through PI3K/Akt activation in OSCC[J]. Stomatology, 2023, 43(5): 393-399.

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