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Gene silencing of Rac1 with RNA interference mediated by ultrasound and microbubbles in human LoVo cells: evaluation of cell invasion inhibition and metastatic
Authors:Fang Nie  Shi-Yi Zhao  Lan Bu  Xue-Hui Liu
Institution:1. Department of Medical Ultrasonics, Lanzhou University Second Hospital, Lanzhou, Gansu, China and;2. Department of Oncology, The Third people’s Hospital of Jinan, Jinan, Shandong, China
Abstract:Objective: The objective of this study was to assess the change of cytoskeleton and cell cycle in LoVo (human colorectal cancer) cell via gene silencing of Rac1 with RNA interference mediated by microbubble (SonoVue) and ultrasound (US).

Methods: The compound of plasmid Rac1-shRNA, LoVo cells, and SonoVue was exposed to US (1?MHz, 2?W/cm2, 5?min). The expression of Rac1 mRNA and Rac1 protein was detected by RT-PCR and Western blot. Cytoskeleton was taken by confocal microscope in a random fashion. Cell invasion was assayed using modified Boyden chambers, and cell cycle and apoptosis were analyzed by flow cytometry in LoVo cells.

Results: Rac1 gene is overexpressed in human colorectal cancer cells, gene silencing of Rac1 with RNA interference mediated by microbubble and US strongly inhibited lamellipodia formation, cell invasion, and delayed cell cycle, as well as enhanced cell apoptosis of LoVo cells in vitro.

Conclusion: Silencing Rac1 gene mediated by microbubble and US may become a new treatment option for the inhibition of the invasion and metastasis of colorectal cancer cells.
Keywords:Actin cytoskeleton  invasion colorectal cancer  microbubble and ultrasound  Rac1  RNA interference
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