ERK信号通路参与至真方逆转人大肠癌多药耐药细胞HCT-8/VCR耐药作用机制的研究

梁伟; 陈志霞; 王慧君; 王松坡

doi:10.3969/j.issn.1671-038X.2015.05.04

ERK信号通路参与至真方逆转人大肠癌多药耐药细胞HCT-8/VCR耐药作用机制的研究

- 1 上海交通大学附属第一人民医院中医科, 上海 200080;
- 2 斜土社区卫生服务中心, 上海 200032
基金项目:
上海市中医药事业发展三年行动计划资助(No:ZYSNXD-CC-HPGC-JD-006)

详细信息

作者简介:
梁伟,女,硕士研究生

通讯作者: 王松坡,E-mail,13386259791@163.com

中图分类号: R735.3

收稿日期: 2014-12-03

Research on the role of ERK signaling pathway in the reversing effect of Zhizhen Recipe on multidrug resistance in human colorectal cancer cell lines

- 1 Department of TCM, the First People's Hospital Affiliated to Shanghai Jiaotong University, Shanghai 200080, China;
- 2 Xietu Community Health Service Center, Shanghai 200032, China

More Information

Corresponding author: WANG Song-po, E-mail: 13386259791@163.com

Received Date: 03 December 2014

摘要

摘要: [目的]探讨至真方是否通过影响ERK通路活性,下调P-gp蛋白及ERK、MDR1 mRNA表达,逆转人大肠癌多药耐药细胞HCT-8/VCR多药耐药。[方法]细胞增殖-毒性检验CCK-8(cell counting kit-8)法鉴定HCT-8/VCR细胞株的多药耐药性及存活率;流式细胞仪测定细胞内罗丹明123(Rh123)的平均荧光强度;Real-time PCR及Western blot检测ERK、p-ERK、P-gp的基因和蛋白的表达。[结果]HCT-8/VCR为多药耐药细胞;至真方含药血清可明显抑制HCT-8/VCR细胞生长;HCT-8/VCR细胞经8%、16%、32%至真方含药血清干预48 h后,细胞内Rho123外排减少,波峰右移,细胞内荧光强度明显增强(P＜0.01);HCT-8/VCR细胞ERK、MDR1 mRNA的表达水平均有所下降,且呈浓度依赖性;p-ERK表达较用药前(0.764±0.001)明显下降(P＜0.01),分别为(0.513±0.002)、(0.498±0.001)、(0.471±0.12);ERK表达较用药前(0.771±0.204)分别下降至(0.437±0.004)、(0.413±0.002)、(0.398±0.001),(P＜0.01)。P-gp表达较用药前(0.94±0.014)下降至(0.701±0.01)、(0.663±0.01)、(0.508±0.02),(P＜0.01)。[结论]至真方可逆转人大肠癌多药耐药细胞株HCT-8/VCR的耐药作用,其机制与降低P-gp外排功能、降低ERK通路活性、下调P-gp蛋白及基因表达有关。
- 大肠癌 /
- 多药耐药 /
- 至真方 /
- ERK通路 /
- P-糖蛋白
Abstract: [Objective] To investigate the role of ERK signaling pathway in the reversing effect of Zhizhen Recipe on multidrug resistance in human colorectal cancer cell lines.[Methods] CCK-8 assay was used to detect the drug resistance of HCT-8/VCR cells and apotosis rate. FCM was used to detect intracellular concentration of Rho123. Real-time PCR and western blot were adopted to detect the protein levels and gene levels of ERK, P-ERK and P-gp.[Results] Compared with sensitive cells,HCT-8/VCR appeared multidrug resistant. The drug-serum containing Zhizhen Recipe significantly inhibited the growth of HCT-8/VCR cells. After treatment with 8%, 16% and 32% of the drug serum for 48 hours,the discharge of intracellular Rho123 by HCT-8/VCR decreased,wave crest was shifted to right, and intracellular fluorescence intensity was strengthened. The expression of ERK and DR1 mRNA decreased, which was correlated to the concentration. Compared with before treatment,expression of p-ERK was markedly decreased respectively from(0.764±0.001)to(0.513±0.002),(0.498±0.001), and(0.471±0.12)(P＜0.01). The expression of ERK was decreased respectively from (0.771±0.204)to(0.437±0.004),(0.413±0.002), and(0.398±0.001)(P＜0.01). The expression of P-gp was markedly decreased respectively from(0.94±0.014)decreased to(0.701±0.01),(0.663±0.01), and(0.508±0.02)(P=0.000).[Conclusion] Drug-serum containing Zhizhen Recipe could reverse the MDR of colon cancer cells. The possible mechanism was by inhibiting the efflux function of P-gp,reducing the activity of ERK pathway,and down regulating the expression of P-gp protein and gene.
- Zhizhen Recipe /
- colorectal cancer /
- MDR /
- ERK /

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