芸香苷逆转人大肠癌LoVo/5-氟尿嘧啶细胞耐药性及其机制

doi:10.16098/j.issn.0529-1356.2020.06.015

解剖学报 ›› 2020, Vol. 51 ›› Issue (6): 906-911.doi: 10.16098/j.issn.0529-1356.2020.06.015

芸香苷逆转人大肠癌LoVo/5-氟尿嘧啶细胞耐药性及其机制

王婷婷¹刘超怡² 李秀芬³ 李娜^1*刘安丽¹

1.郑州澍青医学高等专科学校组织胚胎学与病理学教研室，郑州 450064; 2.黄河科技学院微生物学免疫学教研室，郑州 450003; 3.河南应用技术职业学院基础医学教研室，郑州 450003

收稿日期:2019-05-09 修回日期:2019-11-01 出版日期:2020-12-06 发布日期:2020-12-06
通讯作者: 李娜 E-mail:2877987015@qq.com
基金资助:
河南省高等学校青年骨干教师培养项目

Reversal effect of rutin on drug resistance in human colorectal cancer LoVo/5-fluorouracil cells and its mechanisms

WANG Ting-ting¹ LIU Chao-yi² LI Xiu-fen³ LI Na^1* LIU An-li¹

1.Department of Histoembryology and Pathology, Zhengzhou Shuqing Medical College, Zhengzhou 450064, China; 2.Department of Microbiology and Immunology, Huanghe Science and Technology University, Zhengzhou 450003, China; 3.Department of Basic Medicine, He’nan Vocational College of Applied Technology, Zhengzhou 450003, China

Received:2019-05-09 Revised:2019-11-01 Online:2020-12-06 Published:2020-12-06
Contact: LI Na E-mail:2877987015@qq.com

摘要/Abstract

摘要：

目的探讨芸香苷（RT）对大肠癌耐药株LoVo/5-氟尿嘧啶（5-FU）细胞耐药性的影响及其机制。方法采用浓度梯度递增法建立耐药株LoVo/5-FU细胞；用不同剂量的RT和（或）5-FU处理48 h后，采用细胞计数试剂盒8（CCK-8）法检测细胞活力，流式细胞术检测细胞凋亡和细胞周期，RT-PCR法检测P-糖蛋白（P-gp）和多药耐药相关蛋白1（MRP1）的mRNA表达水平，Western blotting法检测P-gp、MRP1、磷酸化蛋白激酶B（p-Akt）、Akt、Survivin、细胞周期蛋白A（cyclin A）和细胞周期素依赖性蛋白激酶2（CDK2）的蛋白表达水平。结果 LoVo/5-FU细胞中P-gp、MRP1和Survivin蛋白表达水平均显著高于LoVo细胞；5-FU和RT对LoVo/-FU细胞的半数抑制浓度（IC₅₀）分别为21.77 mg/L和98.43 mg/L；在10 mg/L RT作用下，5-FU对LoVo/5-FU细胞的IC₅₀降至10.64 mg/L，逆转倍数为2.05；RT可协同增强5-FU引起的LoVo/5-FU细胞凋亡和S期阻滞，抑制P-gp和MRP1基因表达以及Akt磷酸化，下调Survivin、cyclin A和CDK2蛋白水平。结论 RT可通过抑制Akt信号通路以及耐药相关蛋白的表达反转LoVo/5-FU细胞对5-FU的耐药性。

关键词: 芸香苷, 5-氟尿嘧啶, 大肠癌LoVo/5-FU细胞系, 逆转耐药, 免疫印迹法, 人

Abstract:

Objective To investigate the effects of rutin (RT) on 5-fluorouracil (5-FU) resistance in human colorectal cancer LoVo/5-FU cells and its possible mechanism. Methods The drug-resistant LoVo/5-FU cells were established by stepwise exposure to 5-FU. LoVo and LoVo/5-FU cells were cultured in vitro and treated with RT and（or）5-FU for 48 hours, and then the cell viability were detected by cell counting kit-8(CCK-8) assay. The cell apoptosis and cell cycle were analyzed by flow cytometry. The mRNA expression levels of P-glycoprotein (P-gp) and multidrug resistance associated protein 1 (MRP1) were measured by RT-PCR. The protein levels of P-gp, MRP1, phosphorylated protein kinase B(p-Akt), Survivin, cyclin A and cyclin dependent kinase 2 (CDK2) were measured by Western blotting. Results LoVo/5-FU cells had a significantly higher protein levels of P-gp, MRP1 and survivin than LoVo cells. The 50% inhibitory concentration(IC₅₀) value of LoVo/5-FU cells to 5-FU and RT was 21.77 mg/L and 98.43 mg/L respectively. Under the influence of RT (10 mg/L), the IC₅₀ value of LoVo/5-FU cells to 5-FU reduced to 10.64 mg/L with a reversal fold of 2.05. RT resulted in an enhance of 5-FU-induced apoptosis and S phase arrest, inactivation of Akt, and lower expression of P-gp, MRP1, survivin, cyclin A and CDK2 in LoVo/5-FU cells. Conclusion RT can reverse the drug resistance of LoVo/5-FU cells through down-regulating the expression of drug-resistant related protein and suppressing Akt signaling pathway.

Key words: Rutin, 5-fluorouracil, Colorectal cancer LoVo/5-FU cell, Reversal of drug resistance, Western blotting, Human

中图分类号:

R735.3

王婷婷刘超怡李秀芬李娜刘安丽. 芸香苷逆转人大肠癌LoVo/5-氟尿嘧啶细胞耐药性及其机制[J]. 解剖学报, 2020, 51(6): 906-911.

WANG Ting-ting LIU Chao-yi LI Xiu-fen LI Na LIU An-li. Reversal effect of rutin on drug resistance in human colorectal cancer LoVo/5-fluorouracil cells and its mechanisms[J]. Acta Anatomica Sinica, 2020, 51(6): 906-911.

参考文献

［1］ Nafees S, Mehdi SH, Zafaryab M, et al. Synergistic interaction of rutin and silibinin on human colon cancer cell line［J］. Arch Med Res, 2018, 49(4):226-234.

［2］ Ben Sghaier M, Pagano A, Mousslim M, et al. Rutin inhibits proliferation, attenuates superoxide production and decreases adhesion and migration of human cancerous cells［J］. Biomed Pharmacother, 2016, 84(2016):1972-1978.

［3］ Elsayed HE, Ebrahim HY, Mohyeldin MM, et al. Rutin as a novel c-Met inhibitory lead for the control of triple negative breast malignancies［J］. Nutr Cancer, 2017, 69(8): 1256-1271.

［4］ Martínez Conesa C, Vicente Ortega Ⅴ, Yá?ez Gascón MJ, et al. Treatment of metastatic melanoma B16F10 by the flavonoids tangeretin, rutin, and diosmin［J］. J Agric Food Chem, 2005, 53(17):6791-6797.

［5］ Mohana S, Ganesan M, Rajendra Prasad N, et al. Flavonoids modulate multidrug resistance through wnt signaling in P-glycoprotein overexpressing cell lines［J］. BMC Cancer, 2018, 18(1):1168.

［6］ Iriti M, Kubina R, Cochis A, et al. Rutin, a quercetin glycoside, restores chemosensitivity in human breast cancer cells［J］. Phytother Res, 2017, 31(10): 1529-1538.

［7］ Bourogaa E, Bertrand J, Despeaux M, et al. Hammada scoparia flavonoids and rutin kill adherent and chemoresistant leukemic cells ［J］. Leuk Res, 2011, 35(8):1093-1101.

［8］ Shan JZ, Xuan YY, Zhang Q, et al. Ursolic acid sensitized colon cancer cells to chemotherapy under hypoxia by inhibiting MDR1 through HIF-1α［J］. J Zhejiang Univ Sci B, 2016, 17(9):672-682.

［9］ La X, Zhang L, Li Z, et al. (-)-Epigallocatechin gallate (EGCG) enhances the sensitivity of colorectal cancer cells to 5-FU by inhibiting GRP78/NF-κB/miR-155-5p/MDR1 pathway［J］. J Agric Food Chem, 2019, 67(9):2510-2518.

［10］ Pei G, Luo M, Ni X, et al. Autophagy facilitates metadherin-induced chemotherapy resistance through the AMPK/ATG5 pathway in gastric cancer［J］. Cell Physiol Biochem, 2018, 46(2):847-859.

［11］ Ke M, Dong J, Wang Y, et al. MEL-pep, an analog of melittin, disrupts cell membranes and reverses 5-fluorouracil resistance in human hepatocellular carcinoma cells［J］. Int J Biochem Cell Biol, 2018, 101(2018):39-48.

［12］ Sun L, Ke J, He Z, et al. HES1 promotes colorectal cancer cell resistance to 5-Fu by inducing Of EMT and ABC transporter proteins［J］. J Cancer, 2017, 8(14):2802-2808.

［13］ Liu B, Liu Y, Zhao L, et al. Upregulation of microRN-135b and microRNA-182 promotes chemoresistance of colorectal cancer by targeting ST6GALNAC2 via PI3K/AKT pathway［J］. Mol Carcinog, 2017, 56(12):2669-2680.

［14］ Wang G, Wang JJ, Du L, et al. Inhibitory kinetics and mechanism of flavonoids extracted from cotinus coggygria scop. Against Glioblastoma Cancer［J］. Nutr Cancer, 2016, 68(8):1357-1368.

［15］ Tong L, Wang LJ, Yuan L. The enhancing effect of gambogic acid on the sensibility in human gastric carcinoma SGC7901/DDP cells to cisplatin［J］. Acta Anatomica Sinica, 2018, 49(3): 337-341. (in Chinese)

仝雷, 王丽君, 袁磊. 藤黄酸增强人胃癌SGC7901/DDP细胞对顺铂的敏感性［J］. 解剖学报, 2018, 49(3): 337-341.

［16］ Wu J, Wang L, Du X, et al. α-solanine enhances the chemosensitivity of esophageal cancer cells by inducing microRNA-138 expression［J］. Oncol Rep, 2018, 39(3):1163-1172.

［17］ Ng L, Chow AKM, Man JHW, et al. Suppression of Slit3 induces tumor proliferation and chemoresistance in hepatocellular carcinoma through activation of GSK3β/β-catenin pathway［J］. BMC Cancer, 2018, 18(1):621.

［18］ Kim EJ, Kang GJ, Kang JI, et al. Over-activation of AKT signaling leading to 5-Fluorouracil resistance in SNU-C5/5-FU cells［J］. Oncotarget, 2018, 9(28):19911-19928.

［19］ Wang T, Liu Z, Zhang Z, et al. Evaluation of antitumor activity of survivin short interfering RNA delivered by lipid nanoparticles in colon cancer in vitro and in vivo［J］. Oncol Lett, 2017, 14(2):2001-2008.

［20］ Xu JM, Azzariti A, Tommasi S, et al. Combination of 5-fluorouracil and irinotecan on modulation of thymidylate synthase and topoisomerase Ⅰ expression and cell cycle regulation in human colon cancer LoVo cells: clinical relevance［J］. Clin Colorectal Cancer, 2002, 2(3):182-188.

［21］ Lv M, Li Y, Tian X, et al. Lentivirus-mediated knockdown of NLK inhibits small-cell lung cancer growth and metastasis ［J］. Drug Des Devel Ther, 2016, 10:3737-3746.

［22］ Zou J, Li S, Chen Z, et al. A novel oral camptothecin analog, gimatecan, exhibits superior antitumor efficacy than irinotecan toward esophageal squamous cell carcinoma in vitro and in vivo ［J］. Cell Death Dis, 2018, 9(6): 661.

[1]	洪晓敏李三强崔钦奕郑润月杨孟利罗仁利李前辉 . 酒精性肝纤维化核因子κB信号通路与性别的差异 [J]. 解剖学报, 2024, 55(1): 55-61.
[2]	魏茜茜李超红赵晨露唐家萍刘玉珍. 大鼠颈上神经节中Ⅰ组代谢型谷氨酸受体表达及慢性间歇性低氧对其的影响 [J]. 解剖学报, 2024, 55(1): 3-9.
[3]	袁蕾杨智伟杨惠刘政海何洁万炜. 三七总皂苷抑制小鼠星形胶质细胞活化缓解完全弗氏佐剂所致炎性痛 [J]. 解剖学报, 2024, 55(1): 25-31.
[4]	马婷婷陈建红刘爱翠李海宁. 抑制lncRNA TUG1下调核苷酸结合寡聚结构域样受体蛋白1炎症小体在延缓阿尔茨海默病进展的作用 [J]. 解剖学报, 2024, 55(1): 32-42.
[5]	邹成功冯浩陈兵唐辉邵川孙谋杨荣何家全. 创伤性颅脑损伤中神经功能障碍与认知功能损伤的动态变化 [J]. 解剖学报, 2024, 55(1): 43-48.
[6]	吕海燕沈西雅赵富生朱梅. 松茸多糖对 1-甲基-4-苯基-吡啶离子诱导 PC12 细胞损伤的影响 [J]. 解剖学报, 2024, 55(1): 49-54.
[7]	郑丽平刘霞杜晓华吴亚娟刘珊珊 . 脑源性神经营养因子在牦牛与黄牛端脑不同区域的表达与分布 [J]. 解剖学报, 2024, 55(1): 10-16.
[8]	郝永明郭磊周程辉裴汉军李莉赵哲 . 改良腹主动脉缩窄法建立心肌肥厚模型[J]. 解剖学报, 2024, 55(1): 120-124.
[9]	许亚平余悦欣陈金福王柯柯郭志坤 . 高龄大鼠心室肌间质弹性纤维和胶原纤维的结构分布特征及其机制 [J]. 解剖学报, 2023, 54(6): 716-721.
[10]	王文娟丁雨桐苏昊任捷孙艳荣王涵菲陆嘉莉张林倩白钰秦丽华. 低雌激素状态下大鼠海马及纹状体Nogo-A的表达变化[J]. 解剖学报, 2023, 54(6): 620-627.
[11]	张琴杨俊霞蒋青松. 福辛普利对糖尿病视网膜病变小鼠血管紧张素转化酶2和血管内皮生长因子的影响[J]. 解剖学报, 2023, 54(6): 628-634.
[12]	梁盼盼禹爱梅杜静寇文辉王欢欢宋爱霞. 基于c-Jun氨基末端激酶/p38丝裂原活化蛋白激酶信号通路探讨阿魏酸钠对偏头痛大鼠炎症反应的抑制作用[J]. 解剖学报, 2023, 54(6): 652-659.
[13]	徐颖刘斌郑兴何宗钊. 补体C3a受体在盲肠结扎穿孔致脓毒症大鼠认知障碍中的作用及机制 [J]. 解剖学报, 2023, 54(6): 668-675.
[14]	刘哲川李坤马帅男孟家琪王艳芹. 利拉鲁肽对百草枯诱导的小鼠帕金森病模型炎症及线粒体融合/分裂的影响 [J]. 解剖学报, 2023, 54(6): 676-681.
[15]	吴俊波杨杰肖锋李金亮 . 微小RNA-138调控Wnt通路对体外人脑胶质瘤细胞生物学行为的影响 [J]. 解剖学报, 2023, 54(6): 682-688.

芸香苷逆转人大肠癌LoVo/5-氟尿嘧啶细胞耐药性及其机制

Reversal effect of rutin on drug resistance in human colorectal cancer LoVo/5-fluorouracil cells and its mechanisms

PDF

可视化

摘要/Abstract

引用本文

使用本文

参考文献

相关文章 15

编辑推荐

Metrics

本文评价