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siRNA silencing CCL18 expression inhibits invasion and migration of ovarian epithelial carcinoma SKOV3 cells
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Abstract:
Objective:To investigate the effect of in vitro silencing CCL18 gene expression on invasion and metastasis of ovarian epithelial carcinoma SKOV3 cells. Methods: Three pairs of siRNAs targeting CCL18 (CCL18-siRNA61, CCL18-siRNA127, CCL18-siRNA224) were chemically synthesized, and were transfected into CCL18-positive SKOV3 cells in vitro. RT-PCR was used to detect the expression of CCL18 mRNA in SKOV3 cells. The siRNA sequence with the best interference efficiency was selected to construct interference plasmid targeting CCL18 , named pSilencer4.1-CCL18-siRNA61. After transfection of pSilencer4.1-CCL18-siRNA61 plasmid, the proliferation of SKOV3 cells was detected by MTT assay, the cell cycle was detected by flow cytometry, and the cell invasion, migration and adhesion capacity in vitro was determined by Transwell assay, Migration assay and Fibronectin adhesion method, respectively. Results: CCL18-siRNA61 showed the best interference efficiency in the three CCL18-siRNAs. The interference plasmid pSilencer4.1-CCL18-siRNA61 was then successfully constructed. The expression of CCL18 mRNA was significantly decreased after pSilencer4.1-CCL18-siRNA61 transfection. The proliferation of SKOV3 cells was not affected by pSilencer4.1-CCL18-siRNA61 transfection. However, the proportion of SKOV3 cells in phase (S+G2+M) in the pSilencer4.1-CCL18-siRNA61 transfection group was significantly lower than that in the pSilencer4.1-Ctrl-siRNA transfection group (\[1971±44\]% vs \[26.45±7.91\]%, P<0.05). The invasion, migration and adhesion capacity of SKOV3 cells was effectively inhibited in the pSilencer4.1-CCL18-siRNA61 transfection group compared with that in the pSilencer4.1-Ctrl-siRNA transfection group (\[9.91±3.41\]% vs \[23.75±6.81\]%, \[16.80±8.71\]% vs \[31.74±11.23\]%, \[6.73±4.33\]% vs \[17.53±6.54\]%, P<0.05), respectively. Conclusion: siRNA silencing CCL18 expression can inhibit the invasion, adhesion and migration capacity of ovarian epithelial cancer cell line SKOV3.
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Project Supported:
Project supported by the Key Research Project of Science and Technology Bureau of Guangxi Zhuang Autonomous Region(No. 2010GXNSFD013053), and the Key Research Program of Science and Technology Bureau of Guangxi Zhuang Autonomous Region(No. 1140003A-34)
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