[Effects of RhoA silencing on proliferation of tongue squamous cancer cells].
OBJECTIVE
This study investigated the effect of RhoA silencing through RNA interference on proliferation and growth of tongue cancer cells, as well as explored the possible mechanisms of this effect.


METHODS
SSC-4 tongue cancer cells were cultured in vitro and then transfected with small interfering RNA to knock down RhoA expression. The tested cells were divided into three groups: experimental group (experimental group 1: transfected with RhoA-siRNA-1; experi-mental group 2: transfected with RhoA-siRNA-2), negative control group (transfected by random sequence NC-siRNA), and blank control group (transfected with Lipofectamine). The expression levels of RhoA mRNA were respectively measured by quantitative real-time polymerase chain reaction and western blot assay. Moreover, the expression levels of cyclin D1, p21, and p27 and RhoA protein were evaluated by Western blot assay. Proliferation and growth potentiality were analyzed through evaluation of doubling times and methyl thiazolyl tetrazolium assessment.


RESULTS
The expression levels of RhoA gene and protein of experimental groups significantly decreased following siRNA transfection compared with those in the negative and blank control groups. The expression of cyclin D1 decreased significantly and that of p21 and p27 increased significantly. The doubling time was extended and the growth potentiality decreased.


CONCLUSIONS
The results indicated that RhoA silencing can inhibit proliferation of tongue cancer cells, whereas RhoA affects cell proliferation by regulating the cell cycle pathway. Thus, RhoA is a potential target in gene therapy for tongue cancer.
Dept. of Stomatology, Wuxi No 2. People's Hospital, Wuxi 214002, China.
Yan
Y
Guoxin
Dept. of Stomatology, Wuxi No 2. People's Hospital, Wuxi 214002, China.
Fan
F
Bing
Dept. of Stomatology, Wuxi No 2. People's Hospital, Wuxi 214002, China.
Zou
Z
Ronghai
Dept. of Stomatology, Wuxi No 2. People's Hospital, Wuxi 214002, China.
Zhang
Z
Jian
Dept. of Stomatology, Wuxi No 2. People's Hospital, Wuxi 214002, China.
Sun
S
Xiaofeng
Center of Stomatology, Qingdao Municipal Hospital Affiliated to Qingdao University Medical College, Qingdao 266071, China.
Tong
T
Lei
Center of Stomatology, Qingdao Municipal Hospital Affiliated to Qingdao University Medical College, Qingdao 266071, China.
Wang
W
Qimin
Center of Stomatology, Qingdao Municipal Hospital Affiliated to Qingdao University Medical College, Qingdao 266071, China.
Han
H
Jinhong
Dept. of Stomatology, Pudong Healthcare Center of Jimo County, Qingdao 266234, China.
Lu
L
Xufei
College of Stomatology, Weifang Medical University, Weifang 261021, China.
Wang
W
Ying
College of Stomatology, Weifang Medical University, Weifang 261021, China.
Zhou
Z
Yuan
Center of Stomatology, Qingdao Municipal Hospital Affiliated to Qingdao University Medical College, Qingdao 266071, China.
He
H
Zongxuan
Center of Stomatology, Qingdao Municipal Hospital Affiliated to Qingdao University Medical College, Qingdao 266071, China.
Liao
L
Yixiang
Postgraduate School, Dalian Medical University, Dalian 116044, China.
Li
L
Ning
Postgraduate School, Dalian Medical University, Dalian 116044, China.
Cao
C
Lei
Center of Stomatology, Qingdao Municipal Hospital Affiliated to Qingdao University Medical College, Qingdao 266071, China;Dept. of Oral and Maxillofacial Surgery, Shanghai Ninth People's Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China.
Chen
C
Zhenggang
D045744:Cell Line, Tumor; D049109:Cell Proliferation; D019938:Cyclin D1; D020868:Gene Silencing; D006801:Humans; D018307:Neoplasms, Squamous Cell; D012333:RNA, Messenger; D034741:RNA, Small Interfering; D060888:Real-Time Polymerase Chain Reaction; D014062:Tongue Neoplasms; D014162:Transfection; D020742:rhoA GTP-Binding Protein
C530104:CCND1 protein, human; D012333:RNA, Messenger; D034741:RNA, Small Interfering; C066737:RHOA protein, human; D019938:Cyclin D1; D020742:rhoA GTP-Binding Protein
RNA interference; RhoA; cell proliferation; tongue cancer