%0 Journal Article
%T Effect of SOX2 gene on the biological characteristics of salivary adenoid cystic carcinoma cells.
%A Wei Z
%A Zhou H
%A Fei W
%A Nie X
%A Li M
%J Hua Xi Kou Qiang Yi Xue Za Zhi
%V 40
%N 3
%D 2022 May 25
%M 38597017
暂无%R 10.7518/hxkq.2022.03.015
%X 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).
CONCLUSIONS: 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.
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。.