Establishment of drug-resistant esophageal cancer cell line（Eca-109/VCR）and investigation on its drug-resistance mechanism

doi:10.16155/j.0254-1793.2021.03.09

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Chinese Journal of Pharmaceutical Analysis ›› 2021, Vol. 41 ›› Issue (3) : 446-451. DOI: 10.16155/j.0254-1793.2021.03.09

Metabolism Analysis

Establishment of drug-resistant esophageal cancer cell line（Eca-109/VCR）and investigation on its drug-resistance mechanism

SUN Xiao-ran¹, XIN Yun-chao¹, SUN Jian-jing¹, LI Duo¹, ZHANG Lin-xi²

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Abstract

Objective: To establish a vincristine-resistant cell line of esophageal cancer（Eca-109/VCR）and to investigate the drug-resistance mechanism. Methods: The vincristine（VCR）-resistant esophageal cancer cell line（Eca-109/VCR）was constructed by high-dose intermittent shock combined with time-increasing methods. The morphology of Eca-109/VCR cells were observed under the inverted microscope. The growth curves of Eca-109 and Eca-109/VCR were plotted by cell counting method, and the doubling time was calculated. The IC₅₀ and resistance index of VCR/5-FU/PTX were detected by CCK-8 method. The cell cycle and apoptosis were detected using flow cytometry. The expressions of P-glycoprotein（P-gp）, and multidrug resistance related protein（MRP） were detected by immunocytochemical staining and Western blot. Results: Eca-109/VCR cells were successfully constructed after 6 months. Eca-109/VCR cells mostly grew in clusters. The resistance index to VCR was 17. 60, and presenting multidrug resistance, the resistance index to 5-Fu and PTX were 6. 59 and 5. 60 respectively （P<0. 01）. Compared with Eca-109 cells, the growth doubling time of Eca-109/VCR cells were prolonged;migration and invasion capability were enhanced, and the distribution of cell cycle was changed, as the cell number of G0/G1 and S were observed increased, and that of G2/M phase decreased;the apoptosis rate was lower;Immunocytochemistry and Western blot showed that the relative expression of P-gp and MRP in Eca-109/VCR was higher than that in Eca109 cells（P<0. 05）. Conclusion: Eca-109/VCR cell line has been successfully established. The mechanism of multiple drug-resistance may be related to the increased expression of P-gp and MRP proteins.

Key words

esophageal cancer / Eca-109/VCR cell line / Multiple drug-resistace / Flow cytometry / Western blot

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SUN Xiao-ran, XIN Yun-chao, SUN Jian-jing, LI Duo, ZHANG Lin-xi. Establishment of drug-resistant esophageal cancer cell line（Eca-109/VCR）and investigation on its drug-resistance mechanism[J]. Chinese Journal of Pharmaceutical Analysis, 2021, 41(3): 446-451 https://doi.org/10.16155/j.0254-1793.2021.03.09

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