SIRT1在5-FU耐药胃癌细胞中的表达及其对细胞化疗耐药的影响  

Expression of SIRT1 in 5-FU Resistant Gastric Cancer Cells and Its Role in Chemoresistance

作　　者：邓晓晶[1] 郑海伦[1] 燕善军[1] 李大鹏[1] DENG Xiao-jing;ZHENG Hai-lun;YAN Shan-jun;LI Da-peng(Department of Gastroenterology,The First Affiliated Hospital of Bengbu Medical College,Bengbu,Anhui 233004,China)

机构地区：[1]蚌埠医学院第一附属医院消化内科,安徽蚌埠233004

出　　处：《河北北方学院学报（自然科学版）》2020年第10期1-7,11,共8页Journal of Hebei North University:Natural Science Edition

基　　金：安徽省高校自然科学研究重点项目(No.KJ2019A0336)。

摘　　要：目的探讨沉默信息调节因子1(silent information regulator 1,SIRT1)在5-氟尿嘧啶(5-fluorouracil,5-FU)耐药胃癌细胞(SGC-7901/5-FU)中的表达情况及其对细胞化疗耐药的影响。方法通过定量RT-PCR和Western blot方法检测SIRT1 mRNA与蛋白在5-FU敏感的SGC-7901亲本细胞、SGC-7901/5-FU细胞及在SIRT1-siRNA转染后的SGC-7901/5-FU细胞中的表达;选择高干扰效率的SIRT1-siRNA转染SGC-7901/5-FU细胞,CCK8法检测5-FU对转染细胞的活性影响,流式细胞术检测其对细胞凋亡的影响,Western blot法检测其对细胞p-Akt、Bax、Bcl-2和P-pg表达的影响。结果SGC-7901/5-FU细胞中的SIRT1 mRNA与蛋白表达水平均显著高于SGC-7901亲本细胞(P<0.001);SIRT1-siRNA转染SGC-7901/5-FU细胞后,SIRT1的mRNA与蛋白表达均显著降低(P<0.001);5-FU作用后,SIRT1-siRNA转染组的SGC-7901/5-FU细胞增殖活性明显下降(P<0.001),细胞凋亡明显增加(P<0.001),且细胞中的p-Akt、Bcl-2和P-pg含量明显下降(P<0.001),Bax含量明显上升(P<0.001)。结论SIRT1在5-FU耐药胃癌细胞中高表达,干扰SIRT1表达后可通过抑制细胞增殖与促进细胞凋亡来提高耐药胃癌细胞对5-FU的敏感性,这可能与细胞增殖通路相关蛋白、凋亡以及细胞多药耐药性相关蛋白变化相关。提示抑制SIRT1可作为降低胃癌多药耐药性及提高胃癌对化疗药物敏感性研究的一个新思路。Objective To explore the expression of silent information regulator 1(SIRT1)in 5-FU drug-resistant gastric cancer cells(SGC-7901/5-FU)and its role in chemoresistance.Methods The expression level of SIRT1 mRNA and protein in SGC-7901 parental cells,SGC-7901/5-FU cells and SGC-7901/5-FU cells with transfection of SIRT1-siRNA was detected by quantitative real-time PCR and Western-blot.SIRT1-siRNA with high interfering efficiency was selected to transfect SGC-7901/5-FU cells.The effect of 5-FU on the activity of transfected cells was detected by CCK8,the effect of 5-FU on cell apoptosis was detected by flow cytometry,the effect of 5-FU on protein expression levels of p-AKT,Bax,Bcl-2 and P-pg was detected by Western-blot.Results The mRNA and protein expression level of SIRT1 in SGC-7901/5-FU cells was significantly higher than that in SGC-7901 parental cells(P<0.001).After SIRT1-siRNA transfection into SGC-7901/5-FU cells,the SIRT1 mRNA and protein expression level significantly decreased than before(P<0.001).Compared with the control group,when transfected cells were treated with 5-FU,the cell proliferation activity significantly decreased(P<0.001)and apoptosis increased(P<0.001),the protein expression of p-AKT,Bcl-2 and P-pg in the cells significantly decreased(P<0.001),while Bax expression significantly increased(P<0.001).Conclusion SIRT1 is highly expressed in 5-FU resistant gastric cancer cells,and the sensitivity of SIRT1-siRNA cells to 5-FU can be improved by inhibiting cell proliferation and promoting cell apoptosis after interference with SIRT1 expression in cells,which may be affected by proteins related to cell proliferation pathway,apoptosis and cellular multi-drug resistance.The results suggest that SIRT1 inhibition could be used as a new way to reduce chemoresistance and improve the sensitivity of chemotherapy drugs in gastric cancer.

关 键 词：沉默信息调节因子1 SGC-7901/5-FU SIRT1-siRNA 化疗耐药 胃癌 

分 类 号：R735.2[医药卫生—肿瘤]