RESUMO
Background and purpose:The expression ofRab11 gene was increased incervical cancer cell and may be involved in the cellular malignant transformation. This study used the sequence-speciifc siRNA knocking down the expression of Rab11 gene and aimed to investigate its effect on invasion and migration of cervical cancer cell lines HeLa/SiHa and its mechanism.Methods:HeLa/SiHa cells were divided into 2 groups: non-speciifc siRNA group transfected with unrelated siRNA (Rab11-NC) and Rab11 siRNA group transfected with Rab11 siRNA (Rab11siRNA). Western blot was used to examine the Rab11 protein expression. Cell migration and invasion were detected by cell scratch and Transwell invasion assay. Western blot was used to further investigate the expression of Rac1, matrix metal-loproteinase 2 (MMP2) and MMP9 which were critical for regulating cell invasion. Moreover, immunolfuorescence was used to identify intracellular location of Rac1 in HeLa/SiHa cells.Results:The Rab11 siRNA inhibited expression of Rab11 gene (P<0.01). The invasion and migration capacities of HeLa/SiHa cells were markedly inhibited in Rab11siR-NA group (P<0.05). The expression of Rac1 signiifcantly decreased (P<0.01). The expression of MMP2 and MMP9 de-creased (P<0.05) as well. The recruitment of Rac1 to protruding edge signiifcantly decreased following down-regulation of Rab11.Conclusion:Down-regulatedRab11 expression could inhibit the expression of Rac1, MMP2 and MMP9, and alter the location of Rac1, leading to suppression of HeLa/SiHa cells migration and invasion.
RESUMO
Objective To explore the effects of Rab11 on biological functions of human cervical cancer cell line HeLa through regulating the expression levels of Rab11. Methods The Rab11 siRNA was transfected into HeLa cells and the expression of Rab11 was detected by Western blot. CCK8 assay, colony formation experiments, EdU assay and Transwell assay were adopted to observe the effect of Rab11 on HeLa cells proliferation and invasion. Results The expression of Rab11 was decreased significantly in HeLa cells transfected with Rab11 siRNAs than that in control siRNA (1.096 ±0.091 vs 1.735 ±0.084, P< 0.01). The proliferation was markedly inhibited in Rab11 siRNA group compared with that in control siRNA group (48 h:0.721±0.092 vs 1.090±0.099; 72 h: 0.956±0.105 vs 1.482±0.096; 96 h: 1.231±0.099 vs 1.720±0.174, P< 0.01), the number of colonies was lower than that in control siRNA group (36±1 vs 75±8, P< 0.01) and so was proliferation rate [(33.880±1.902) % vs (45.570±2.025) %, P< 0.05]. The cell invasion rate of Rab11 siRNA group was lower than that of control siRNA group [(38.6 ±0.8) % vs (100.0 ±0.2) %, P< 0.01]. Conclusion Down-regulation of Rab11 expression can inhibit the growth of HeLa cells.