School of Stomatology, Qingdao University, Qingdao 266003, China.
Dept. of Stomatology, Qingdao Municipal Hospital, Qingdao University, Qingdao 266071, China.
Hua Xi Kou Qiang Yi Xue Za Zhi. 2021 Oct 1;39(5):510-517. doi: 10.7518/hxkq.2021.05.003.
This study aims to investigate the effect of RhoE expression on the migration and invasion of tongue squamous cell carcinoma (TSCC).
Forty-eight TSCC cases were selected from the Maxillofacial Surgery Center of Qingdao Municipal Hospital from 2017 to 2019. The expression of RhoE in the specimens (TSCC and adjacent tissues) was detected by immunohistochemistry, and RhoE mRNA and protein were extracted to further detect the expression of RhoE. SCC-4 and CAL-27 cells were selected for experiments. Transient transfection was used to overexpress RhoE. Real-time fluorescence quantitative PCR (qRT-PCR) and Western blot analyses were conducted to detect the overexpression efficiency. Scratch test and Transwell cell invasion tests were used to detect the migration and invasion ability of TSCC, respectively. The expression levels of Rho-associated coiled-coil-containing protein kinase 1 (ROCK1), matrix metalloproteinase-2 (MMP-2), and matrix metalloproteinase-9 (MMP-9) were detected by Western blot. Experimental data were analyzed by Graphpad prism 8.2.1 software.
The expression level of RhoE in TSCC was significantly lower than that in adjacent tissues (<0.05). The migration and invasion abilities of TSCC were significantly lower than those in the control group (<0.05). The Western blot showed significantly lower expression levels of ROCK1, MMP-2, and MMP-9 in the experimental group than in the control group (<0.05).
RhoE expression is low in TSCC. Over expression RhoE in TSCC can significantly decrease its migration and invasion abilities. Hence, RhoE may play an important role in regulating the metastasis and invasion of TSCC and provide a new target for gene therapy.
本研究旨在探讨 RhoE 表达对舌鳞状细胞癌(TSCC)迁移和侵袭的影响。
选取 2017 年至 2019 年青岛市市立医院颌面外科中心 48 例 TSCC 患者,采用免疫组化法检测标本(TSCC 组织及癌旁组织)中 RhoE 的表达,提取 RhoE mRNA 和蛋白,进一步检测 RhoE 的表达。选取 SCC-4 和 CAL-27 细胞进行实验,瞬时转染过表达 RhoE,实时荧光定量 PCR(qRT-PCR)和 Western blot 分析检测 RhoE 的过表达效率,划痕实验和 Transwell 细胞侵袭实验分别检测 TSCC 的迁移和侵袭能力,Western blot 检测 Rho 相关卷曲螺旋蛋白激酶 1(ROCK1)、基质金属蛋白酶-2(MMP-2)和基质金属蛋白酶-9(MMP-9)的表达水平。Graphpad prism 8.2.1 软件分析实验数据。
TSCC 中 RhoE 的表达水平明显低于癌旁组织(<0.05)。TSCC 的迁移和侵袭能力明显低于对照组(<0.05)。Western blot 显示实验组 ROCK1、MMP-2 和 MMP-9 的表达水平明显低于对照组(<0.05)。
RhoE 在 TSCC 中表达较低,过表达 RhoE 可显著降低 TSCC 的迁移和侵袭能力,提示 RhoE 可能在调节 TSCC 的转移和侵袭中发挥重要作用,为基因治疗提供新的靶点。