FGFR3 silencing by siRNA inhibits invasion of A549 cells
Yuhua Li1, Xiguang Liu1, Hongjun Zhang1
1Department of Oncology, The Affiliated Hospital of Qingdao University, Qingdao, Shandong 266003, P.R. China.
Abstract:
The present study identified that fibroblast growth factor receptor 3 (FGFR3) was significantly upregulated in bone metastasis of lung adenocarcinoma. RNA interference (RNAi) is a powerful approach for treating a wide range of human diseases, including cancer, through downregulating the expression of selected genes. In the present study, the invasiveness of A549 cells cultured in vitro was altered by small interfering (si)RNA targeting FGFR3, and the regulatory effect of silencing FGFR3 on the expression levels of E-cadherin and matrix metalloproteinase (MMP)9 was investigated. Human lung adenocarcinoma A549 cells were transfected with synthetic specific siRNAs targeting a fragment of the FGFR3 gene (namely, siRNA-855, siRNA-1447 and siRNA-2076) or with negative control (NC) siRNA. Cells were divided into five groups (A, siRNA-855 group; B, siRNA-1447 group; C, siRNA-2076 group; D, NC-siRNA group; and E, blank control group). The effect of the above siRNAs targeting FGFR3 on the invasion capacity of A549 cells was detected by Transwell assay. siRNAs against FGFR3 were transfected into A549 cells with by Lipofectamine® 2000, and the expression levels of FGFR3, E-cadherin and MMP9 were measured by reverse transcription-quantitative polymerase chain reaction and western blot assay. The experimental findings indicated that the expression levels of FGFR3 and MMP9 were significantly reduced in the siRNA-FGFR3-transfected groups (A-C groups), compared with those in the D and E groups (P<0.01). In addition, the expression levels of E-cadherin were markedly elevated in the A-C groups, compared with those in the D and E groups (P<0.01). There was no significant difference in E-cadherin expression between the A-C groups, or between the D and E groups (P>0.05). These results indicated that siRNA-FGFR3 was able to decrease the invasiveness of A549 cells, inhibit the expression of MMP9 and increase the expression of E-cadherin by downregulating the expression of FGFR3. Taken together, the results of the present study indicated that the upregulation of E-cadherin expression and the downregulation of MMP9 expression are able to inhibit the migration of A549 cells, and siRNA silencing FGFR3 acts as a tumor suppressor in these cells.
Insights
RNA interference targeting fibroblast growth factor receptor 3 (FGFR3) reduced lung adenocarcinoma cell invasion. Silencing FGFR3 decreased matrix metalloproteinase 9 and increased E-cadherin expression, acting as a tumor suppressor.
Area of Science:
- Oncology
- Molecular Biology
- Gene Therapy
Background:
- Fibroblast growth factor receptor 3 (FGFR3) is upregulated in lung adenocarcinoma bone metastasis.
- RNA interference (RNAi) offers a potential therapeutic strategy for cancer by downregulating gene expression.
Purpose of the Study:
- To investigate the effect of silencing FGFR3 using small interfering RNA (siRNA) on the invasiveness of A549 lung adenocarcinoma cells.
- To determine the regulatory impact of FGFR3 silencing on E-cadherin and matrix metalloproteinase (MMP)9 expression.
Main Methods:
- A549 cells were transfected with specific siRNAs targeting FGFR3 or a negative control (NC) siRNA.
- Cell invasiveness was assessed using Transwell assays.
- Expression levels of FGFR3, E-cadherin, and MMP9 were quantified via RT-qPCR and Western blot.
Main Results:
- Transfection with siRNA targeting FGFR3 significantly reduced FGFR3 and MMP9 expression.
- E-cadherin expression was markedly elevated in cells treated with siRNA targeting FGFR3.
- FGFR3 silencing decreased A549 cell invasiveness.
Conclusions:
- Silencing FGFR3 inhibits lung adenocarcinoma cell invasion by downregulating MMP9 and upregulating E-cadherin.
- FGFR3 acts as a tumor suppressor in A549 cells through modulation of these key proteins.
- RNAi targeting FGFR3 presents a potential therapeutic approach for lung adenocarcinoma metastasis.
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