酸性培养对人舌鳞癌细胞增殖、凋亡、迁移能力的影响及其机制研究

doi:10.7518/hxkq.2021.05.007

华西口腔医学杂志 ›› 2021, Vol. 39 ›› Issue (5): 540-546.doi: 10.7518/hxkq.2021.05.007

酸性培养对人舌鳞癌细胞增殖、凋亡、迁移能力的影响及其机制研究

代晓华(), 王冠华, 连小丽, 颜艳, 王悦, 邹慧儒, 刘浩()

天津市口腔功能重建重点实验室天津市口腔医院南开大学医学院，天津 300041

收稿日期:2020-09-07 修回日期:2021-07-03 出版日期:2021-10-01 发布日期:2021-10-11
通讯作者: 刘浩 E-mail:jstonehome@163.com;kqlh2013@163.com
作者简介:代晓华，副研究员，硕士，E-mail：jstonehome@163.com
基金资助:
天津市临床医学重点学科专项(HWZX001);天津市科技计划项目(19ZXDBSY00070);天津市自然科学基金(18JC-YBJC27000);天津市口腔功能重建重点实验室项目(2021KL-MS13)

Effect of acidic culture conditions on the proliferation, apoptosis, and migration ability of human tongue squamous cell carcinoma cells and its related mechanism

Dai Xiaohua(), Wang Guanhua, Lian Xiaoli, Yan Yan, Wang Yue, Zou Huiru, Liu Hao.()

Tianjin Key Laboratory of Oral and Maxillofacial Function Reconstruction, Tianjin Stomatological Hospital, School of Medicine, Nankai University, Tianjin 300041, China

Received:2020-09-07 Revised:2021-07-03 Online:2021-10-01 Published:2021-10-11
Contact: Liu Hao. E-mail:jstonehome@163.com;kqlh2013@163.com
Supported by:
Special Project of Tianjin Clinical Medicine Key Discipline(HWZX001);Tianjin Science and Technology Projects(19ZXDBSY00070);Natural Science Foundation of Tianjin City of China(18JCYBJC27000);Project of Tianjin Key Laboratory of Oral and Maxillofacial Function Reconstruction(2021KLMS13);Correspondence: Liu Hao, E-mail: kqlh2013@163.com

摘要/Abstract

摘要： 目的

探究酸性培养条件对人舌鳞癌细胞SCC15和CAL27增殖、凋亡、迁移能力的影响及潜在的分子机制。

方法

在pH6.2的酸性培养液中分别培养一定时间，噻唑兰（MTT）法检测舌鳞癌细胞SCC15和CAL27增殖能力；流式细胞分析法检测SCC15和CAL27细胞凋亡水平的改变；划痕愈合实验检测SCC15和CAL27迁移能力改变；实时荧光定量聚合酶链反应（FQ-PCR）检测酸性培养后SCC15和CAL27细胞中COX-2与survivin基因表达情况。

结果

酸性培养24 h的SCC15和CAL27细胞生长曲线显示，细胞酸性培养后，经历2~3 d生长相对缓慢的调整期，之后快速增长达到生长平衡期。酸性培养后SCC15和CAL27细胞凋亡水平降低，以酸性培养6 h细胞凋亡率下降最为显著。酸性培养后SCC15和CAL27细胞划痕愈合率提高。FQ-PCR检测结果显示酸性培养后SCC15和CAL27细胞中COX-2和survivin表达均升高。

结论

酸性培养可抑制舌鳞癌细胞凋亡，促进其迁移，诱导出适应性更强、恶性程度更高的舌鳞癌细胞，其机制可能与COX-2和survivin及其涉及的信号通路有关。

关键词: 舌鳞癌, 酸性微环境, 细胞迁移, 细胞凋亡

Abstract: Objective

This study aims to explore the effect of acidic culture conditions on the proliferation, apoptosis, and migration ability of human tongue squamous cell carcinoma SCC15 and CAL27 cells and its potential molecular mechanism.

Methods

After acidic culture for different periods, methyl thiazolyl tetrazolium (MTT) method was adop-ted to detect the cell proliferation of SCC15 and CAL27. Flow cytometry was employed to detect the apoptosis level of SCC15 and CAL27 cells. The migration ability of SCC15 and CAL27 after acidic culture was detected by scratch hea-ling test. Real-time fluorescence quantitative polymerase chain reaction (FQ-PCR) was used to detect the mRNA expression of cyclooxygenase 2 (COX-2) and survivin in SCC15 and CAL27 cells after acidic culture.

Results

After culture for 24 h under acidic microenvironment, SCC15 and CAL27 cells grew rapidly and reached the stationary phase after adjustment for 3 days. The apoptosis levels of SCC15 and CAL27 cells decreased after acidic culture, but the most significant reduction occurred after 6 h of acidic culture. The scratch healing rates of SCC15 and CAL27 cells increased after acidic culture. The results of FQ-PCR showed that the mRNA expression levels of COX-2 and survivin in SCC15 and CAL27 cells increased after acidic culture.

Conclusion

Extracellular acidic microenvironment can inhibit the apoptosis of tongue squamous carcinoma cells, promote their migration, and induce more adaptable and malignant tongue squamous carcinoma cells. The mechanism may be related to COX-2 and survivin and their signal pathways.

Key words: tongue squamous cell carcinoma, acidic extracellular environment, cell migration, cell apoptosis

中图分类号:

R 739.8

代晓华, 王冠华, 连小丽, 颜艳, 王悦, 邹慧儒, 刘浩. 酸性培养对人舌鳞癌细胞增殖、凋亡、迁移能力的影响及其机制研究[J]. 华西口腔医学杂志, 2021, 39(5): 540-546.

Dai Xiaohua, Wang Guanhua, Lian Xiaoli, Yan Yan, Wang Yue, Zou Huiru, Liu Hao.. Effect of acidic culture conditions on the proliferation, apoptosis, and migration ability of human tongue squamous cell carcinoma cells and its related mechanism[J]. West China Journal of Stomatology, 2021, 39(5): 540-546.

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酸性培养对人舌鳞癌细胞增殖、凋亡、迁移能力的影响及其机制研究

Effect of acidic culture conditions on the proliferation, apoptosis, and migration ability of human tongue squamous cell carcinoma cells and its related mechanism

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