SOX2基因对唾液腺腺样囊性癌细胞生物学特性的影响

doi:10.7518/hxkq.2022.03.015

华西口腔医学杂志 ›› 2022, Vol. 40 ›› Issue (3): 341-349.doi: 10.7518/hxkq.2022.03.015

SOX2基因对唾液腺腺样囊性癌细胞生物学特性的影响

魏卓¹^,²(), 周昊¹, 费伟¹^,²(), 聂雄², 李明芳²

^1.四川省医学科学院四川省人民医院口腔科，成都 610072
^2.川北医学院口腔医学院，南充 637000

收稿日期:2021-05-27 修回日期:2021-10-14 出版日期:2022-06-01 发布日期:2022-06-01
通讯作者: 费伟 E-mail:2456018637@qq.com;1264954098@qq.com
作者简介:魏卓，硕士，E-mail：2456018637@qq.com
基金资助:
四川省科技计划项目(2017JY0082);四川省人民医院院基金重点项目(2021LZ01)

Effect of SOX2 gene on the biological characteristics of salivary adenoid cystic carcinoma cells

Wei Zhuo¹^,²(), Zhou Hao¹, Fei Wei¹^,²(), Nie Xiong², Li Mingfang²

^1.Dept. of Stomatology, Sichuan Academy of Medical Sciences & Sichuan Provincial People’s Hospital, Chengdu 610072, China
^2.School of Stomatology, North Sichuan Medical College, Nanchong 637000, China

Received:2021-05-27 Revised:2021-10-14 Online:2022-06-01 Published:2022-06-01
Contact: Fei Wei E-mail:2456018637@qq.com;1264954098@qq.com
Supported by:
Science and Technology Project of Sichuan Province(2017JY0082);Key Project of Sichuan Provincial People’s Hospital Foundation(2021LZ01);Correspondence: Fei Wei,E-mail: 1264954098@qq.com

摘要/Abstract

摘要： 目的

探索转染SOX2-shRNA载体慢病毒对唾液腺腺样囊性癌（SACC）细胞系SACC-LM和SACC-83生物学行为的影响。

方法

构建3种SOX2-shRNA慢病毒载体（817、818、819）并将其转染到SACC细胞中，通过实时荧光定量聚合酶链反应（RT-qPCR）和蛋白质印迹（Western blot）筛选出抑制效果最好的shRNA。将抑制效果最好的慢病毒载体转染到SACC-LM、SACC-83细胞后，Western blot检测SOX2、Survivin、E-钙黏蛋白（E-cadherin）、N-钙黏蛋白（N-cadherin）的表达变化，CCK-8和流式细胞术检测细胞增殖和凋亡情况，细胞划痕实验及Transwell法检测细胞的迁移和侵袭能力。

结果

3种慢病毒中SOX2-shRNA-819干扰效果最好。SACC-LM、SACC-83细胞转染SOX2-shRNA-819慢病毒后，SOX2、Survivin、N-cadherin的表达下调，E-cadherin的表达上调（P<0.05）；细胞增殖能力降低，凋亡能力增强（P<0.05）；细胞迁移和侵袭能力下降（P<0.05）。

结论

shRNA干扰技术降低SOX2表达的同时下调了SACC-LM中Survivin的表达，二者的表达可能具有相关性；SOX2的低表达抑制了SACC细胞的增殖、迁移和侵袭能力，促进了凋亡能力。SOX2可能参与了SACC上皮间充质转化过程，为靶向SOX2基因治疗SACC提供了相关的理论依据。

关键词: 唾液腺腺样囊性癌, SOX2, RNA干扰, 生物学行为, 增殖, 迁移, 侵袭, 凋亡, 慢病毒

Abstract: Objective

This study aimed to investigate the effect of transfecting SOX2-shRNA vector lentivirus to SACC cell lines on the biological behavior of salivary adenoid cystic carcinoma (SACC)-LM and SACC-83.

Methods

Three types of SOX2-shRNA lentiviral vectors (817, 818, and 819) were constructed and transfected successfully. The shRNA with the best inhibitory effect was screened out and transfected into SACC cells. Real-time fluorescence quantitative polymerase chain reaction (RT-qPCR) and Western blot were used to detect the expressions of Survivin, E-cadherin, and N-cadherin of SACC-LM and SACC-83. CCK-8 and flow cytometry were used to detect SACC-LM and SACC-83 proliferation and apoptosis. Cell scratch test and Transwell method were used to detect the migration and invasion capabilities of SACC-LM and SACC-83.

Results

SOX2-shRNA-819 had the best interference effect among the three lentiviruses. After transfecting SOX2-shRNA-819 into SACC-LM and SACC-83, the expressions of SOX2, Survivin, and N-cadherin were significantly reduced, and that of E-cadherin was significantly increased (P<0.05). The cell proliferation ability decreased, and the number of apoptotic cells increased (P<0.05). The cell migration and invasion ability decreased (P<0.05).

Conclusion

shRNA interference technology reduced SOX2 expression while downregulating Survivin expression. These two expressions may be related. Low SOX2 expression inhibits the proliferation, migration, and invasion of SACC cells and promotes the apoptosis of SACC cells. SOX2 may be involved in the epithelial⁃mesenchymal transition process. This study provided a relevant theoretical basis for targeting SOX2 gene therapy in SACC.

Key words: salivary adenoid cystic carcinoma, SOX2, RNA interference, biological behavior, proliferation, migration, invasion, apoptosis, lentivirus

中图分类号:

R 739.8

魏卓, 周昊, 费伟, 聂雄, 李明芳. SOX2基因对唾液腺腺样囊性癌细胞生物学特性的影响[J]. 华西口腔医学杂志, 2022, 40(3): 341-349.

Wei Zhuo, Zhou Hao, Fei Wei, Nie Xiong, Li Mingfang. Effect of SOX2 gene on the biological characteristics of salivary adenoid cystic carcinoma cells[J]. West China Journal of Stomatology, 2022, 40(3): 341-349.

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慢病毒载体	靶点序列
SOX2-shRNA-817	5’-CGCTCATGAAGAAGGATAAG-3’
SOX2-shRNA-818	5’-TGGACAGTTACGCGCACATG-3’
SOX2-shRNA-819	5’-CAGCTCGCAGACCTACATGA-3’
Negative-shRNA-376	5’-CGAGACACGAACGGAGAAGA-3’

组别	SOX2	E-cadherin	N-cadherin	Survivin
Mock组	1.110±0.105	0.646±0.049	1.102±0.035	1.170±0.057
pUc组	1.023±0.120	0.653±0.043	1.058±0.029	1.085±0.064
pU组	0.507±0.012^ab	0.792±0.050^ab	0.513±0.014^ab	0.631±0.096^ab

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SOX2基因对唾液腺腺样囊性癌细胞生物学特性的影响

Effect of SOX2 gene on the biological characteristics of salivary adenoid cystic carcinoma cells

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