[1]林增海,陆 军,王凯松.槲皮素对5-FU诱导的结直肠癌SW480细胞耐药及自噬调控机制研究[J].陕西中医,2021,(10):1338-1343.[doi:DOI:10.3969/j.issn.1000-7369.2020.10.004]
　LIN Zenghai,LU Jun,WANG Kaisong.Regulatory mechanism of quercetin on resistance and autophagy of 5-FU induced colorectal cancer SW480 cells[J].,2021,(10):1338-1343.[doi:DOI:10.3969/j.issn.1000-7369.2020.10.004]
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槲皮素对5-FU诱导的结直肠癌SW480细胞耐药及自噬调控机制研究
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《陕西中医》[ISSN:1000-7369/CN:61-1281/TN]

卷:
期数:
2021年10期
页码:
1338-1343
栏目:
基础研究
出版日期:
2021-10-05

文章信息/Info

Title:
Regulatory mechanism of quercetin on resistance and autophagy of 5-FU induced colorectal cancer SW480 cells
作者:
林增海; 陆 军; 王凯松
(汕头大学医学院第一附属医院肝胆外科,广东 汕头 515041)
Author(s):
LIN Zenghai; LU Jun; WANG Kaisong
(Department of Hepatobiliary Surgery,the First Affiliated Hospital of Shantou University Medical College,Shantou 515041,China)
关键词:
结直肠癌;  槲皮素;  5-氟尿嘧啶;  细胞耐药;  自噬;  丝/苏氨酸激酶;  雷帕霉素靶蛋白
Keywords:
Colorectal cancer;  Quercetin;  5-fluorouracil;  Cell resistance;  Autophagy;  Serine/threonine kinase;  Mammalian target of rapamycin
分类号:
R 735.4
DOI:
DOI:10.3969/j.issn.1000-7369.2020.10.004
文献标志码:
A
摘要:
目的:探讨槲皮素(Que)调控5-氟尿嘧啶(5-FU)诱导的结直肠癌SW480细胞耐药及自噬的作用及机制。方法:采用浓度递增诱导法建立5-FU耐药细胞株SW480/5-FU,MTT法设置Que高、中、低剂量组(10、20、40 μmol/L)和对照组。MTT法检测各组细胞的5-FU耐药性,TUNEL染色法检测细胞凋亡,免疫荧光检测细胞自噬活性,Western blot检测细胞p糖蛋白(Pgp)、多药耐药相关蛋白1(MRP1)、三磷酸腺苷结合转运蛋白G超家族成员2抗体(ABCG2)、LC3Ⅰ、LC3Ⅱ、p62、丝/苏氨酸激酶(AKT)、p-AKT、雷帕霉素靶蛋白(mTOR)和p-mTOR蛋白表达。结果:5-FU抑制SW480/5-FU细胞的半抑制浓度(IC50)明显高于SW480细胞(P<0.05),耐药指数(RI)=13.74。与对照组相比,Que中、高剂量组5-FU抑制SW480/5-FU细胞的IC50明显下降(P<0.05),耐药逆转倍数分别为2.27和4.03。与对照组相比,Que中、高剂量组细胞抑制率、凋亡率和自噬活性明显升高(均P<0.05),蛋白的表达均明显升高(均P<0.05),P-gp、MRP1、ABCG2、p62、p-AKT/AKT和p-mTOR/mTOR蛋白表达明显下降(均P<0.05)。结论:Que可以以剂量依赖性的方式逆转SW480/5-FU细胞的5-FU耐药性,诱导细胞自噬,其作用机制可能与抑制AKT/mTOR的磷酸化有关。
Abstract:
Objective:To explore the effects and regulatory mechanism of quercetin(Que)on resistance and autophagy of 5-fluorouracil(5-FU)induced colorectal cancer(CRC)SW480 cells.Methods:5-FU drug-resistant cell line SW480/5-FU was established by concentration increasing method.The high-dose,medium-dose and low-dose Que groups(10,20,40 μmol/L),and control group were set up by MTT.The resistance of 5-FU in each group was detected by MTT.The cells apoptosis was detected by TUNEL staining.The autophagy activity of cells was detected by immunofluorescence.The expression of p glycoprotein(Pgp),multidrug resistance associated protein 1(MRP1),ATP-binding transporter G cassette transporer G2(ABCG2),LC3Ⅰ,LC3Ⅱ,p62,serine/threonine kinase(AKT),p-AKT,mammalian target of rapamycin(mTOR)and p-mTOR was detected by Western blot.Results:The half maximal inhibitory concentration(IC50)of 5-FU for SW480/5-FU cells was significantly higher than that for SW480 cells(P<0.05),and the resistance index(RI)was 13.74.Compared with control group,the IC50 of 5-FU in SW480/5-FU cells was significantly decreased in medium-dose and high-dose Que groups(P<0.05),and the resistance reversal folds were 2.27 and 4.03,respectively.Compared with control group,inhibition rate,apoptosis rate and autophagy activity of cells were significantly increased in medium-dose and high-dose Que groups(all P<0.05),expression of LC3Ⅱ/LC3Ⅰ was significantly increased(all P<0.05),and expression of P-gp,MRP1,ABCG2,p62,p-AKT/AKT and p-mTOR/mTOR was significantly decreased(all P<0.05).Conclusion:Que can reverse 5-FU resistance of SW480/5-FU cells with dose-dependence,and induce autophagy.Its mechanism of action may be related to inhibiting AKT/mTOR phosphorylation.

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备注/Memo

备注/Memo:
基金项目:广东省自然科学基金资助项目(2018A030307045); 广东省汕头市科技计划项目(180404094011033)
更新日期/Last Update: 2021-10-09