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RNA interfering of CTSL expression inhibits invasion and migration of ovarian cancer cells
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Abstract:
Objective:To construct a cathepsin L (CTSL)-siRNA eukaryotic expression plasmid and investigate its influence on the invasion and migration of ovarian cancer cells. Methods: Four pairs of small interfering RNA sequences targeting CTSL were designed and synthesized, and were transfected into ovarian cancer A2780 cells. CTSL expressions were analyzed by RT-PCR in different CTSL-siRNA transfected-A2780 cells, and the siRNA pair with the best interfering effect was chosen; the corresponding CTSL-shRNA was synthesized and inserted into psilence4.1-CMV-neo plasmid to construct psilence4.1-CTSL plasmid. Psilence4.1-CTSL was then transfected into A2780 cells and the stable transfectant A2780-CTSL was obtained. The interfering effect of psilence4.1-CTSL was detected by RT-PCR and Western blotting analysis; the proliferation of A2780 cells was examined by MTT and colony formation assay; the cell cycle of A2780 cells was measured by flow cytometry; and the invasion and migration of A2780 cells were detected by Transwell chamber assay. Results: The siRNA-CTSL-1202 sequence with the best interfering effect was selected and the corresponding CTSL-shRNA expression plasmid psilence4.1-CTSL was successfully constructed. CTSL expression in psilence4.1-CTSL-stably transfected A2780 cells was significantly decreased. The invasion and migration of A2780 cells were inhibited by CTSL silence, while their proliferation, cell cycle and adhesion were not significantly influenced. Conclusion: The siRNA eukaryotic expression plasmid targeting CTSL gene is successfully constructed, and interfering CTSL expression can suppress invasion and migration of ovarian cancer cells.
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Project Supported:
Project supported by the Public Health Bureau of Guangxi Zhuang Autonomous Region of China （No. 200614）
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