[性别决定区Y盒9下调对口腔鳞状癌细胞上皮间质转化及克隆的影响]

[Effect of down-regulation of sex determining region Y-box 9 on epithelial mesenchymal transition and cloning of oral squamous carcinoma cells].

作者信息

Yang Wen-Li, Sun Ming-Lei, Zhang Peng, Yu Wei-Wei, Zhou Hai-Xia, Sun Qiang

机构信息

Dept. of Stomatology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, China.

出版信息

Hua Xi Kou Qiang Yi Xue Za Zhi. 2019 Feb 1;37(1):13-18. doi: 10.7518/hxkq.2019.01.003.

DOI:10.7518/hxkq.2019.01.003

PMID:30854812

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC7030727/

Abstract

OBJECTIVE

To investigate the effect of sex determining region Y-box 9 (SOX9) on epithelial mesenchymal transition (EMT) and cloning of oral squamous cell carcinoma (OSCC).

METHODS

siRNA control, SOX9 siRNA were transfected into BcaCD885 cells in OSCC. Simultaneously, cells that did not undergo transfection were used as the control. Quantitative real time polymerase chain reaction (qRT-PCR) and Western blot were used to select SOX9 siRNA1 with enhanced interference effect. A cell cloning assay was used to determine the cell's clone formation ability. E-cadherin and Vimentin expressions were detected by immunofluorescence. The expressions of E-cadherin, matrix metalloprotease 2 (MMP-2), Vimentin and matrix metalloprotease 9 (MMP-9) were detected by Western blot. Cell invasion and migration were detected in the Transwell compartment.

RESULTS

The levels of SOX9 mRNA and protein in SOX9 siRNA cells were significantly lower than those of the control (P<0.05). An increase in the number of SOX9 siRNA1 cell clonesled to the considerable decrease of the number of cell invasion and migration. In addition, levels of MMP-2 and MMP-9 proteins in cells decreased significantly compared with the control (P<0.05). The level of Vimentin expression in SOX9 siRNA1 cells decreased, and expression level of E-cadherin was elevated. Cell EMT was inhibited compared with the control, and the difference was statistically significant (P<0.05).

CONCLUSIONS

Down-regulation of SOX9 inhibited EMT, clonogenic formation, cell invasion and OSCC migration.

摘要

目的

探讨Y染色体性别决定区框9（SOX9）对口腔鳞状细胞癌（OSCC）上皮-间质转化（EMT）及克隆形成的影响。

方法

将siRNA对照、SOX9 siRNA转染至OSCC的BcaCD885细胞中。同时，将未进行转染的细胞作为对照。采用定量实时聚合酶链反应（qRT-PCR）和蛋白质免疫印迹法筛选干扰效果增强的SOX9 siRNA1。采用细胞克隆实验检测细胞的克隆形成能力。通过免疫荧光检测E-钙黏蛋白和波形蛋白的表达。采用蛋白质免疫印迹法检测E-钙黏蛋白、基质金属蛋白酶2（MMP-2）、波形蛋白和基质金属蛋白酶9（MMP-9）的表达。在Transwell小室中检测细胞侵袭和迁移情况。

结果

SOX9 siRNA细胞中SOX9 mRNA和蛋白水平显著低于对照组（P<0.05）。SOX9 siRNA1细胞克隆数量增加，导致细胞侵袭和迁移数量显著减少。此外，与对照组相比，细胞中MMP-2和MMP-9蛋白水平显著降低（P<0.05）。SOX9 siRNA1细胞中波形蛋白表达水平降低，E-钙黏蛋白表达水平升高。与对照组相比，细胞EMT受到抑制，差异具有统计学意义（P<0.05）。