硼替佐米对高三尖杉酯碱耐药细胞株K562的作用及其机制

张悦; 陈锦豪; 郭宗明; 徐敏

doi:10.13201/j.issn.1004-2806.2021.05.008

硼替佐米对高三尖杉酯碱耐药细胞株K562的作用及其机制

- 张家港市第一人民医院血液科(江苏张家港,215600)
基金项目:
苏州市科技发展计划项目(No:SYSD2015006)

国家自然科学基金青年基金项目(No:81800205)

详细信息

通讯作者: 徐敏,E-mail:xmzjgws@163.com

中图分类号: R733.72

收稿日期: 2021-02-23

Effects and mechanisms of bortezomib on homoharringtonine-resistant cell line K562

- Department of Hematology, Zhangjiagang First People's Hospital, Zhangjiagang, 215600, China

More Information

Corresponding author: XU Min, E-mail: xmzjgws@163.com

Received Date: 23 February 2021

摘要

摘要: 目的:探究硼替佐米对高三尖杉酯碱耐药细胞株K562(K562/HHT)的作用及其机制。方法:采用浓度梯度增加法建立K562/HHT细胞;CCK-8法检测不同浓度硼替佐米分别作用K562和K562/HHT细胞24 h、48 h后对细胞增殖抑制的作用;流式细胞术分别检测硼替佐米单独或联合高三尖杉酯碱作用于K562/HHT细胞48 h后的细胞凋亡;Western blotting技术检测硼替佐米单独或联合高三尖杉酯碱作用K562/HHT细胞48 h后BCL-2、P170蛋白表达的变化。结果:成功建立K562/HHT细胞,24 h耐药倍数为678.31倍,48 h耐药倍数为787.31倍。硼替佐米对K562/HHT细胞的增殖抑制呈浓度、时间依赖性(P<0.05);低浓度硼替佐米能够逆转K562/HHT的耐药性,24 h逆转倍数为2.89倍,48 h逆转倍数为3.85倍。低浓度硼替佐米联合高三尖杉酯碱能够增加K562/HHT的凋亡;低浓度硼替佐米能够抑制K562/HHT细胞P170蛋白的表达,但不影响BCL-2的表达。结论:低浓度硼替佐米可以逆转K562/HHT细胞对高三尖杉酯碱的耐药,增加K562/HHT的化疗敏感性,其机制可能与下调耐药蛋白P170相关。
- 硼替佐米 /
- K562/HHT /
- 耐药 /
- 高三尖杉酯碱
Abstract: Objective: To investigate the effects and mechanisms of bortezomib on homoharringtonine-resistant cell line K562/HHT.Methods: The homoharringtonine-resistant of K562/HHT cell line was cultured in gradient concentrations of homoharringtonine. CCK-8 cell proliferation assay was used to measure the viability of K562 and K562/HHT cells treated by bortezomib for 24 h and 48 h, respectively. The cell apoptosis of K562/HHT cells after treating with bortezomib alone or combined with homoharringtonine for 48 h was analyzed by flow cytometry. Western blotting was used to detect the protein expressions of BCL-2 and P170.Results: K562/HHT was established successfully, which showed 678.31 folds(24 h) and 787.31 folds(48 h) of homoharringtonine resistance compared with the naive cells. Bortezomib inhibits the proliferation of K562/HHT cells in a concentration-and time-dependent manner(P<0.05); Low doses of bortezomib could reverse K562/HHT resistance, and reverse 2.89 at 24 h and 3.85 at 48 h. Low doses of bortezomib combined with homoharringtonine increased the apoptosis of K562/HHT. Western blotting showed that low doses of bortezomib decreased the protein levels of P170 in K562/HHT cells without affecting BCL-2 protein expression.Conclusion: Low doses of bortezomib can reverse the drug resistance and sensitize homoharringtonine-resistant cell line K562/HHT to chemical drugs possibly by down-regulating the drug-resistant protein P170.
- bortezomib /
- K562/HHT /
- drug resistance /
- homoharringtonine

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