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; 李秀芬; 刘超怡; 李娜; 刘安丽

doi:10.16098/j.issn.0529-1356.2020.06.015

PDF(9067 KB)

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

肿瘤生物学

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

作者信息 +

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

Author information +

文章历史 +

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。

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.

导出引用

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 https://doi.org/10.16098/j.issn.0529-1356.2020.06.015

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 https://doi.org/10.16098/j.issn.0529-1356.2020.06.015

中图分类号： R735.3

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.