下调性别决定区Y框蛋白9对口腔鳞癌细胞上皮间质转化及克隆能力的影响

doi:10.7518/hxkq.2019.01.003

摘要/Abstract

摘要：

目的探讨下调性别决定区Y框蛋白9（SOX9）对口腔鳞癌（OSCC）细胞上皮间质转化（EMT）及克隆能力的影响。方法 OSCC BcaCD885细胞中转染siRNA control、SOX9 siRNA,同时以不做任何转染的细胞作为control组。实时定量聚合酶链式反应（qRT-PCR）和Western blot筛选干扰效果较好的SOX9 siRNA1做后续研究。细胞克隆实验测定细胞克隆形成能力,免疫荧光检测上皮钙黏附素（E-cadherin）、波形蛋白（Vimentin）表达,Western blot检测E-cadherin、基质金属蛋白酶-2（MMP-2）、Vimentin、基质金属蛋白酶-9（MMP-9）表达,Transwell小室检测细胞侵袭和迁移。结果 SOX9 siRNA组细胞中SOX9 mRNA和蛋白水平均明显低于control组（P<0.05）。SOX9 siRNA1细胞克隆形成数目、细胞侵袭和迁移数目明显降低,并且细胞中MMP-2、MMP-9蛋白水平也明显降低,与control组相比,差异具有统计学意义（P<0.05）。SOX9 siRNA1组细胞中Vimentin表达水平降低,而E-cadherin表达水平升高,细胞EMT受到抑制,与control组相比,差异具有统计学意义（P<0.05）。结论下调SOX9可抑制OSCC细胞EMT和克隆形成,以及细胞侵袭和迁移。

关键词: 性别决定区Y框蛋白9, 口腔鳞癌, 上皮间质转化, 细胞克隆

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 trans-fected into BcaCD885 cells in OSCC. Simultaneously, cells that did not undergo transfection were used as the control. Quanti-tative 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), Vi-mentin 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 com-pared with the control, and the difference was statistically significant (P<0.05). Conclusion Down-regulation of SOX9 inhibited EMT, clonogenic formation, cell invasion and OSCC migration.

Key words: sex determining region Y-box 9, oral squa-mous cell carcinoma, epithelial mesenchymal transition, cell clones

中图分类号:

Q785

杨文丽,孙明磊,张朋,余炜伟,周海霞,孙强. 下调性别决定区Y框蛋白9对口腔鳞癌细胞上皮间质转化及克隆能力的影响[J]. 华西口腔医学杂志, 2019, 37(1): 13-18.

Wenli Yang,Minglei Sun,Peng Zhang,Weiwei Yu,Haixia Zhou,Qiang Sun. Effect of down-regulation of sex determining region Y-box 9 on epithelial mesenchymal transition and cloning of oral squamous carcinoma cells[J]. West China Journal of Stomatology, 2019, 37(1): 13-18.

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组别	SOX9 mRNA	SOX9蛋白
control	1.00	0.84±0.06
siRNA control	1.02±0.09^#	0.86±0.07^#
SOX9 siRNA1	0.21±0.05*	0.15±0.02*
SOX9 siRNA2	0.36±0.02*^&	0.29±0.05*^&

组别	E-cadherin	Vimentin
control	0.25±0.06	0.82±0.09
siRNA control	0.22±0.04^#	0.84±0.10^#
SOX9 siRNA1	0.79±0.05*	0.28±0.06*

组别	MMP-2	MMP-9
control	0.81±0.09	0.48±0.04
siRNA control	0.80±0.11^#	0.49±0.06^#
SOX9 siRNA1	0.35±0.04*	0.30±0.05*