- Author:
Jun-Jun SUN
1
;
Yun-Ya TAO
2
;
Yuan ZHOU
2
;
Zong-Xuan HE
3
;
Shan-Gui SHENG
2
;
Qi-Min WANG
2
;
Lei TONG
2
;
Kai ZHAO
2
;
Shao-Ru WANG
4
;
Zheng-Gang CHEN
2
Author Information
- Publication Type:Journal Article
- Keywords: Rce1; invasion; migration; squamous cell carcinoma
- MeSH: Cell Line, Tumor; Cell Movement; Cell Proliferation; Endopeptidases; metabolism; Humans; Neoplasm Invasiveness; RNA Interference; RNA, Small Interfering; Tongue Neoplasms; metabolism; therapy; Transfection
- From: West China Journal of Stomatology 2019;37(2):143-148
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVE:This study aimed to explore the influence of Rce1 on invasion and migration of tongue squamous cell carcinoma cells by silencing the Rce1 gene with RNA interference.
METHODS:The tongue squamous cell carcinoma Cal-27 and SCC-4 cells were cultured in vitro. The small interfering RNA (siRNA) of the Rce1 gene was designed, and the Rcel gene expression was silenced vialiposome transfection. According to the siRNA transfected by liposome, the experimental group was divided into three groups, namely, Rce1-siRNA-1, Rce1-siRNA-2, and Rce1-siRNA-3 groups. Negative control group was transfected by siCON, and the blank control group was untransfected by siRNA. The Rce1, RhoA, and K-Ras gene expression levels in each group were analyzed by real-time quantitative polymerase chain reaction. The Rce1, RhoA, K-Ras, MMP-2, and MMP-9 protein expression levels were analyzed by Western blot. The invasiveness of tongue cancer cell Cal-27 and SCC-4 were determined by Transwell invasion assay, and cell migration assay was performed by cell scratch assay.
RESULTS:Real-time quantitative polymerase chain reaction and Western blot results showed that compared with the negative and blank control groups, the Rce1 gene and protein expression levels in three experimental groups decreased (P<0.05). The RhoA, K-Ras gene and protein expression levels were insignificantly different among groups (P>0.05). Meanwhile, the MMP-2 and MMP-9 expression levels decreased (P<0.05). Transwell invasion assay results showed that the total number of cells in the PET film of the experimental groups was significantly decreased compared with the control group (P<0.05). The cell scratch test showed that the cell closure time of the scratch in the interference group was significantly longer than those in the control and blank groups (P<0.05).
CONCLUSIONS:Silencing Rce1 in vitro can effectively downregulate its expression in tongue squamous cell carcinoma cells Cal-27 and SCC-4 and reduce the migration and invasion abilities of these cells.