Sheng Shan-Gui, Wang Ya-Nan, Wang Shao-Ru, Zhao Kai, Wang Yun-Ying, Xu Xiao-Na, Wang Qi-Min, Tong Lei, Chen Zheng-Gang
School of Stomatology, Qingdao University, Qingdao 266003, China.
Dept. of Oral and Maxillofacial Surgery, Qingdao Stomatological Hospital, Qingdao 266000, China.
Hua Xi Kou Qiang Yi Xue Za Zhi. 2020 Apr 1;38(2):177-184. doi: 10.7518/hxkq.2020.02.012.
This study aimed to explore the effects of silencing farnesyltransferase (FTase) on the migration and invasion of tongue squamous cell carcinoma (TSCC) through RNA interference.
TSCC cells (CAL27 and SCC-4) were cultured in vitro and then transfected with siRNA to silence FTase expression. The tested cells were categorized as follows: experimental group (three RNA interference groups), negative control group, and blank control group. mRNA expression of FTase and HRAS in each group was analyzed by quantitative real-time polymerase chain reaction. On the basis of FTase mRNA expression, the optimum interference group (highest silencing efficiency) was selected as the experimental group for further study. The protein expression of FTase, HRAS, p65, p-p65(S536), matrix metalloprotein-9 (MMP-9), hypoxia inducible factor-1α (HIF-1α), and vascular endothelial growth factor (VEGF) was analyzed by Western blot. The invasion and migration abilities of TSCC cells were determined by Transwell invasion assay and cell wound healing assay.
The mRNA and protein expression of FTase in the experimental group decreased compared with that in the negative control and blank control groups (P<0.05). The mRNA and protein expression of HRAS was not significantly different among the groups (P>0.05). In the experimental group, the protein expression of p-p65(S536), MMP-9, HIF-1α, and VEGF decreased (P<0.05), whereas that of p65 had no significant change (P>0.05). The migration and invasion abilities of the experimental group were inhibited significantly (P<0.05).
Silencing FTase in vitro could effectively downregulate its expression in TSCC cell lines and reduce the migration and invasion abilities to a certain extent. FTase could be a new gene therapy target of TSCC, and this research provided a new idea for the clinical treatment of TSCC.
本研究旨在通过RNA干扰探讨沉默法尼基转移酶（FTase）对舌鳞状细胞癌（TSCC）迁移和侵袭的影响。
体外培养TSCC细胞（CAL27和SCC - 4），然后用小干扰RNA（siRNA）转染以沉默FTase表达。将受试细胞分为以下几类：实验组（三个RNA干扰组）、阴性对照组和空白对照组。通过定量实时聚合酶链反应分析每组中FTase和HRAS的mRNA表达。根据FTase mRNA表达情况，选择最佳干扰组（沉默效率最高）作为实验组进行进一步研究。通过蛋白质免疫印迹法分析FTase、HRAS、p65、磷酸化p65（S536）、基质金属蛋白酶 - 9（MMP - 9）、缺氧诱导因子 - 1α（HIF - 1α）和血管内皮生长因子（VEGF）的蛋白质表达。通过Transwell侵袭实验和细胞划痕愈合实验测定TSCC细胞的侵袭和迁移能力。
与阴性对照组和空白对照组相比，实验组中FTase的mRNA和蛋白质表达降低（P<0.05）。各组之间HRAS的mRNA和蛋白质表达无显著差异（P>0.05）。在实验组中，磷酸化p65（S536）、MMP - 9、HIF - 1α和VEGF的蛋白质表达降低（P<0.05），而p65的蛋白质表达无显著变化（P>0.05）。实验组的迁移和侵袭能力受到显著抑制（P<0.05）。
体外沉默FTase可有效下调其在TSCC细胞系中的表达，并在一定程度上降低迁移和侵袭能力。FTase可能是TSCC的一个新的基因治疗靶点，本研究为TSCC的临床治疗提供了新思路。
