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Generation and characterization of a paclitaxel-resistant human gastric carcinoma cell line

Fu, Linga,b,c,d,*; Yin, Fena,b,c,d,*; Li, Xiao-Ruia,b,c,d,*; Han, Bing-Kaia,b,c,d; Zhang, Chuanga,b,c,d; Wang, Jun-Weia,b,c,d; Wang, Yu-Qinga,b,c,d; Bi, Yue-Fenga,b,c,d; Liu, Hong-Mina,b,c,d

doi: 10.1097/CAD.0000000000000601
PRECLINICAL REPORTS
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The main aim of this study was to establish a novel paclitaxel (PTX)-resistant human gastric carcinoma cell line and to investigate its biological significance. A cell line, MGC803/PTX, was established by gradually increasing PTX density on the basis of MGC803 over a period of 10 months. In addition, a pair of resistant cell lines (SW620 and SW620/PTX) were added to further explain the resistant mechanism of PTX. The drug resistance index and stability of MGC803/PTX cells were detected using the Cell Counting Kit-8 method. The morphological features were observed using inverted microscopy. Apoptosis was measured by flow cytometry (FCM) and Hoechst 33258 fluorescence staining. The distribution of the cell cycle was determined by FCM, and protein expressions of P-gp, Bcl-2, Bax, and PARP were detected by western blot analysis. When characterizing the resistance in vitro, we found that MGC803/PTX cells were 10.3-fold more resistant to PTX compared with MGC803 cells. In addition, MGC803/PTX cells showed cross-resistance to 5-fluorouracil and adriamycin. FCM and Hoechst 33258 fluorescence staining indicated that MGC803/PTX cells had a significantly lower percentage of apoptotic cells after treatment with PTX compared with MGC803 cells. Other differences between parental cells and resistant cells included morphology, proliferation rate, doubling time, cell cycle distribution, and colony-formation rate. Western blot analysis indicated that P-gp, Bcl-2, and PARP protein were more abundant in MGC803/PTX and SW620/PTX cells compared with MGC803 and SW620 cells, whereas Bax protein levels were lower in resistant cells. Furthermore, MGC803/PTX cells showed obvious resistance to PTX in vivo. To our knowledge, this is the first report on the establishment of a PTX-resistant MGC803 cell line, which is an important tool to explore the resistance of anticancer drugs and to overcome tumor drug resistance.

aDepartment of Pharmaceutical Chemistry, School of Pharmaceutical Sciences, Zhengzhou University

bKey Laboratory of Henan Province for Drug Quality and Evaluation

cKey Laboratory of Technology of Drug Preparation, Ministry of Education

dCollaborative Innovation Center of New Drug Research and Safety Evaluation, Zhengzhou, Henan Province, People’s Republic of China

* Ling Fu, Fen Yin and Xiao-Rui Li contributed equally to the writing of this article.

Correspondence to Dr Hong-Min Liu, Department of Pharmaceutical Chemistry, School of Pharmaceutical Sciences, Zhengzhou University, Zhengzhou 450001, Henan Province, China Tel: +86 371 6778 1908; fax: +86 371 6778 1890; e-mail: liuhm@zzu.edu.cn

Received September 27, 2017

Accepted January 11, 2018

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