温州医科大学学报
溫州醫科大學學報
온주의과대학학보
Journal of Wenzhou Medical University
2015年
2期
106-111
,共6页
黄崇杰%刘长宝%郑晨果%蔡锚
黃崇傑%劉長寶%鄭晨果%蔡錨
황숭걸%류장보%정신과%채묘
结直肠肿瘤%热休克蛋白27%5-氟尿嘧啶%RNA干扰
結直腸腫瘤%熱休剋蛋白27%5-氟尿嘧啶%RNA榦擾
결직장종류%열휴극단백27%5-불뇨밀정%RNA간우
colorectal carcinoma%HSP27%5-lfuorouracil%RNA interference
目的：探讨不同大肠癌细胞热休克蛋白（HSP27）蛋白表达水平及其与5-氟尿嘧啶（5-FU）化疗敏感性的关系，同时通过RNA干扰抑制HSP27基因的表达，初步探讨沉默该基因对5-FU抗肿瘤作用的影响。方法：Western blot法检测各大肠癌细胞中HSP27蛋白的表达水平，CCK-8法测定不同浓度5-FU对各细胞株的抑制率，分析各组细胞HSP27蛋白表达水平与IC50的关系。设计合成3对针对不同靶点的HSP27小干扰RNA，通过脂质体转染SW480细胞，激光共聚焦显微镜观察转染效率，Real-time PCR及Western blot法检测转染后HSP27 mRNA及蛋白表达水平。CCK-8法检测HSP27-siRNA对细胞的生长抑制作用及沉默HSP27基因细胞对5-FU化疗敏感性的变化。结果：各组细胞中HSP27的表达水平与5-FU的敏感性呈负相关性。Real-time PCR及Western blot法结果显示有两对靶向HSP27-siRNA能有效抑制HSP27 mRNA及蛋白水平的表达；CCK-8法检测结果显示HSP27-siRNA转染可增强SW480细胞对5-FU的敏感性，半数抑制浓度（IC50）明显下调。HSP27在大肠癌细胞中的表达水平与5-FU化疗药IC50值呈正相关。结论：靶向HSP27的siRNA能有效抑制大肠癌SW480细胞HSP27蛋白的表达，增强5-FU化疗药物的敏感性。
目的：探討不同大腸癌細胞熱休剋蛋白（HSP27）蛋白錶達水平及其與5-氟尿嘧啶（5-FU）化療敏感性的關繫，同時通過RNA榦擾抑製HSP27基因的錶達，初步探討沉默該基因對5-FU抗腫瘤作用的影響。方法：Western blot法檢測各大腸癌細胞中HSP27蛋白的錶達水平，CCK-8法測定不同濃度5-FU對各細胞株的抑製率，分析各組細胞HSP27蛋白錶達水平與IC50的關繫。設計閤成3對針對不同靶點的HSP27小榦擾RNA，通過脂質體轉染SW480細胞，激光共聚焦顯微鏡觀察轉染效率，Real-time PCR及Western blot法檢測轉染後HSP27 mRNA及蛋白錶達水平。CCK-8法檢測HSP27-siRNA對細胞的生長抑製作用及沉默HSP27基因細胞對5-FU化療敏感性的變化。結果：各組細胞中HSP27的錶達水平與5-FU的敏感性呈負相關性。Real-time PCR及Western blot法結果顯示有兩對靶嚮HSP27-siRNA能有效抑製HSP27 mRNA及蛋白水平的錶達；CCK-8法檢測結果顯示HSP27-siRNA轉染可增彊SW480細胞對5-FU的敏感性，半數抑製濃度（IC50）明顯下調。HSP27在大腸癌細胞中的錶達水平與5-FU化療藥IC50值呈正相關。結論：靶嚮HSP27的siRNA能有效抑製大腸癌SW480細胞HSP27蛋白的錶達，增彊5-FU化療藥物的敏感性。
목적：탐토불동대장암세포열휴극단백（HSP27）단백표체수평급기여5-불뇨밀정（5-FU）화료민감성적관계，동시통과RNA간우억제HSP27기인적표체，초보탐토침묵해기인대5-FU항종류작용적영향。방법：Western blot법검측각대장암세포중HSP27단백적표체수평，CCK-8법측정불동농도5-FU대각세포주적억제솔，분석각조세포HSP27단백표체수평여IC50적관계。설계합성3대침대불동파점적HSP27소간우RNA，통과지질체전염SW480세포，격광공취초현미경관찰전염효솔，Real-time PCR급Western blot법검측전염후HSP27 mRNA급단백표체수평。CCK-8법검측HSP27-siRNA대세포적생장억제작용급침묵HSP27기인세포대5-FU화료민감성적변화。결과：각조세포중HSP27적표체수평여5-FU적민감성정부상관성。Real-time PCR급Western blot법결과현시유량대파향HSP27-siRNA능유효억제HSP27 mRNA급단백수평적표체；CCK-8법검측결과현시HSP27-siRNA전염가증강SW480세포대5-FU적민감성，반수억제농도（IC50）명현하조。HSP27재대장암세포중적표체수평여5-FU화료약IC50치정정상관。결론：파향HSP27적siRNA능유효억제대장암SW480세포HSP27단백적표체，증강5-FU화료약물적민감성。
Objective:To investigate the relationship between the HSP27 expression level and the sensiti-zation 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 efifciency was observed by laser confocal mi-croscope. 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 signiifcantly. Conclusion:The corre-lation between the level of HSP27 expression of colon cancer cells and IC50 of 5-FU are positive. HSP27-siRNA leads to the efifcient inhibition of HSP27 expression in SW480 cells, and increase the sensitization to 5-FU.
