ABSTRACT
OBJECTIVES@#This study aims to investigate the effect of RhoE expression on the migration and invasion of tongue squamous cell carcinoma (TSCC).@*METHODS@#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 @*RESULTS@#The expression level of RhoE in TSCC was significantly lower than that in adjacent tissues (@*CONCLUSIONS@#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.
Subject(s)
Humans , Carcinoma, Squamous Cell , Cell Line, Tumor , Matrix Metalloproteinase 2 , Neoplasm Invasiveness , Tongue , Tongue Neoplasms , rho GTP-Binding Proteins/genetics , rho-Associated KinasesABSTRACT
OBJECTIVES@#The effect of isoprenylcysteine carboxymethyltransferase (ICMT) silencing on the migration and invasion of tongue squamous cell carcinoma was investigated by constructing the small interfering RNA (siRNA) of ICMT.@*METHODS@#Through liposomal transfection, siRNA was transfected into human tongue squamous cell carcinoma CAL-27 and SCC-4 cells (ICMT-siRNA group) with a negative control group (transfected with NC-siRNA) and a blank control group (transfected with a transfection reagent but not with siRNA). Quantitative real-time polymerase chain reaction was performed to analyze the mRNA expression of ICMT and RhoA in each group of cells after transfection and to measure the silencing efficiency. Western blot was applied to examine the expression levels of ICMT, total RhoA, membrane RhoA, ROCK1, matrix metalloproteinase (MMP)-2, and MMP-9 proteins in each group. The migration and invasion abilities were evaluated via wound healing and Transwell motility assays.@*RESULTS@#After CAL-27 and SCC-4 cells were transfected with ICMT-siRNA, the expression levels of ICMT genes and proteins decreased significantly in the experimental group compared with those in the negative and blank control groups (@*CONCLUSIONS@#The migration and invasion abilities of CAL-27 and SCC-4 cells were reduced significantly after the transfection of ICMT-siRNA, and the involved mechanism might be related to the RhoA-ROCK signaling pathway.
Subject(s)
Humans , Carcinoma, Squamous Cell , Cell Line, Tumor , Cell Movement , Cell Proliferation , Neoplasm Invasiveness , Protein Methyltransferases , RNA, Small Interfering , Tongue , Tongue Neoplasms , Transfection , rho-Associated KinasesABSTRACT
OBJECTIVES@#This study aimed to explore the effects of silencing isoprenylcysteine carboxyl methyltransfe-rase (Icmt) through small interfering RNA (siRNA) interference on the proliferation and apoptosis of tongue squamous cell carcinoma (TSCC).@*METHODS@#Three siRNA were designed and constructed for the Icmt gene sequence and were then transfected into TSCC cells CAL-27 and SCC-4 to silence Icmt expression. The tested cells were divided as follows: RNA interference groups Icmt-siRNA-1, Icmt-siRNA-2, and Icmt-siRNA-3, negative control group, and blank control group. The transfection efficiency of siRNA was detected by the fluorescent group Cy3-labeled siRNA, and the expression of Icmt mRNA was screened by quantitive real-time polymerase chain reaction (qRT-PCR) selected the experimental group for subsequent experiments. The expression of Icmt, RhoA, Cyclin D1, p21, extracellular regulated protein kinases (ERK), and phospho-extracellular regulated protein kinases (p-ERK) were analyzed by Western blot. The proliferation abilities of TSCC cells were determined by cell counting kit-8 assay. The change in apoptosis was detected by AnnexinV-APC/propidium staining (PI) assay. Cell-cycle analysis was conducted by flow cytometry.@*RESULTS@#The expression of Icmt mRNA and protein in TSCC cells significantly decreased after Icmt-siRNA transfection (@*CONCLUSIONS@#Silencing Icmt can effectively downregulate its expression in TSCC cells, reduce the RhoA membrane targeting localization and cell proliferation, and induce apoptosis. Thus, Icmt may be a potential gene therapy target for TSCC.
Subject(s)
Humans , Apoptosis , Carcinoma, Squamous Cell , Cell Line, Tumor , Cell Proliferation , Protein Methyltransferases , RNA, Small Interfering , Tongue , Tongue NeoplasmsABSTRACT
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 (P0.05). 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.
Subject(s)
Humans , Cell Line, Tumor , Cell Movement , Cell Proliferation , Endopeptidases , Metabolism , Neoplasm Invasiveness , RNA Interference , RNA, Small Interfering , Tongue Neoplasms , Metabolism , Therapeutics , TransfectionABSTRACT
<p><b>OBJECTIVE</b>To establish a novel nude mice model which can be visualized in real time and detected in a continuous and dynamic way for the development and metastasis of adenoid cystic carcinoma.</p><p><b>METHODS</b>Human adenoid cystic carcinoma cells, ACCM cell line, were infected with retroviral vector of pLEGFP-N1 and then screened for a single colony of ACCM-GFP cells. Cell proliferation and morphological analysis were conducted for ACCM and ACCM-GFP cells. Nude mice lingual carcinoma model was set up with ACCM-GFP cells injection and real time observation with fluorescence imaging on ACCM-GFP tumors was performed subsequently. Histological assay was analyzed for ACCM and ACCM-GFP tumors as well.</p><p><b>RESULTS</b>ACCM-GFP cells were able to express GFP stably in the long term. ACCM and ACCM-GFP cells showed no significant difference in cell proliferation and morphology, and no significant difference of histological characteristics in vivo could be found between ACCM and ACCM-GFP tumors. Tumor development could be monitored in real time with fluorescence imaging system in vivo.</p><p><b>CONCLUSION</b>GFP-expressing ACCM tumor model can be applied to detect and observe its development in the long term in a noninvasive, real time and dynamic way. It is also a kind of ideal in vivo mouse model for adenoid cystic carcinoma research.</p>
Subject(s)
Animals , Humans , Mice , Carcinoma, Adenoid Cystic , Cell Line, Tumor , Cell Proliferation , Disease Models, Animal , Green Fluorescent Proteins , Mice, Inbred BALB C , Mice, NudeABSTRACT
OBJECTIVE@#To explore the surgical treatment and effect of intracranial anterior circulation aneurysms.@*METHODS@#Thirty-eight patients with intracranial anterior circulation aneurysms were enrolled, 9 were treated with endovascular embolization,and 29 with pterion approach micro-euthyphoria operation.@*RESULTS@#One patient was postoperative death. Thirty-four patients were followed up. Among them, 26 were recovery, 1 was botan animation, 2 were meta-palsy, 3 oculomotor palsy, and 2 epilepsy.@*CONCLUSION@#Surgical treatment of intracranial anterior circulation aneurysms is the first choice to help blood tumor cleaning-up and intracranial pressure degrading. Embolotherapy can be applied for patients unfit for operation, but we do not recommend wide use of it due to preoperative cranial nerve injury.