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目的:观察Rac1蛋白在结直肠癌细胞中的表达,并分析其与结直肠癌LoVo细胞骨架、细胞周期和细胞凋亡的相关性。方法:Western blotting测定5种结直肠癌细胞株(LoVo,SW480,SW620,SW1116,HT29)中Rac1蛋白的表达;Rac1-shR-NA质粒转染LoVo细胞后,激光共聚焦显微镜观察LoVo细胞骨架的变化,流式细胞仪检测细胞周期和细胞凋亡的变化。结果:5种结直肠癌细胞株中均有Rac1蛋白的高表达。Rac1基因沉默后,LoVo细胞中交联状态的F-actin网明显减少且紊乱,G0/G1期细胞比例较对照组显著增加[(74.63±4.40)%vs(56.46±3.09)%,P<0.05],而S期细胞比例较对照组显著减少[(12.87±1.77)%vs(29.66±1.92)%,P<0.05];Rac1-shRNA转染组LoVo细胞凋亡率较对照组显著增加[(25.31±2.05)%vs(9.38±1.16)%,P<0.05]。结论:RNA沉默Rac1基因的表达影响了LoVo细胞骨架的形成和细胞周期,为以Rac1为靶点治疗结直肠癌提供了实验依据。
OBJECTIVE: To observe the expression of Rac1 protein in colorectal cancer cells and to analyze its correlation with the cytoskeleton, cell cycle and apoptosis of colorectal cancer LoVo cells. Methods: The expression of Rac1 in five colorectal cancer cell lines (LoVo, SW480, SW620, SW1116 and HT29) was detected by Western blotting. The LoVo cells were transfected with LoVo cells by Rac1-shR-NA plasmid. Changes of cell cycle and apoptosis were detected by flow cytometry. Results: Rac1 protein was highly expressed in all five colorectal cancer cell lines. After silencing Rac1 gene, the F-actin network in the LoVo cells was significantly reduced and disorderly compared with the control group [(74.63 ± 4.40)% vs (56.46 ± 3.09)%, P <0.05 (12.87 ± 1.77)% vs (29.66 ± 1.92)%, P <0.05]. The apoptosis rate of LoVo cells in Rac1-shRNA transfected group was significantly increased compared with that in control group [( 25.31 ± 2.05)% vs (9.38 ± 1.16)%, P <0.05]. Conclusion: The expression of RNA silencing Rac1 gene affects the formation and cell cycle of LoVo cytoskeleton and provides an experimental basis for the treatment of colorectal cancer with Rac1 as a target.