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| Silencing gene of heat shock protein 27 increases the sensitivity to 5-FU in colorectal cancer cells |
| Department of Colorectal Surgery, the Second Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325027 |
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HUANG Chongjie,LIU Changbao,ZHENG Chenguo, et al. Silencing gene of heat shock protein 27 increases the sensitivity to 5-FU in colorectal cancer cells[J]. JOURNAL OF WEZHOU MEDICAL UNIVERSITY, 2015, 45(2): 106-.
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Abstract Objective: To investigate the relationship between the HSP27 expression level and the sensitization to 5-FU in different kinds of colon cancer cells, and then explore the effect of HSP27 gene down-regulated by siRNA interference, and the sensitive of 5-FU. Methods: The HSP27 expression level in different kinds of colon cancer cells were detected by Western blot, and the inhibition rates of different concentration of 5-FU were determined by CCK-8 method, then the relationship between the relative level and IC50 of 5-FU was analyzed by Linear correlation. Three siRNA sequences targeted on special sequence of HSP27 gene were designed, All siRNAs were transfected by lipofectamine 2000. The transfection efficiency was observed by laser confocal microscope. The expression of HSP27 was tested with qRT-PCR and Western blot analysis. Different concentrations of 5-FU were added to the cells, which were transfected with siRNA-HSP27 48 h before. Then, the inhibition rate was determined by CCK-8 after 24 h. Results: The correlation between the level of HSP27 expression and IC50 were positive. Real-time PCR and Western blot revealed that two targeted HSP27-siRNA could effectively inhibit the mRNA and protein expression of HSP27. CCK-8 showed that the transfection of HSP27-siRNA can improve the sensitivity of SW480 to 5-FU and the IC50 was downregulated significantly. Conclusion: The correlation between the level of HSP27 expression of colon cancer cells and IC50 of 5-FU are positive. HSP27-siRNA leads to the efficient inhibition of HSP27 expression in SW480 cells, and increase the sensitization to 5-FU.
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Received: 07 August 2014
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2015, Vol. 45 
