紫杉醇诱导子宫内膜癌细胞耐药的生物学特性及机制研究。
Study on Biological Characteristics and Mechanism of Paclitaxel Induced Drug Resistance in Endometrial Carcinoma Cells.
机构信息
The Third Affiliated Hospital, Sun Yat-sen University, Guangdong Province, China.
The Seventh Affiliated Hospital, Sun Yat-sen University, Guangdong Province, China.
出版信息
Biomed Res Int. 2018 Aug 5;2018:8372085. doi: 10.1155/2018/8372085. eCollection 2018.
OBJECTIVE
To study the biological characteristics of paclitaxel resistant endometrial carcinoma cells and its mechanism of drug resistance.
METHOD
The paclitaxel resistant cell lines were established by high-dose paclitaxel (TAX) injection. The IC50 of paclitaxel was determined by CCK-8 assay in Ishikawa and Ishikawa-TAX. The cell cycle and apoptosis rate were detected by flow cytometry. Western blot was used to detect the expression of p-AKT and p-p70S6K. The expression of drug resistance-related genes Pgp and MDR1 was determined by RT-PCR. Cell viability was determined by soft agarose assay and invasive ability in vitro by transwell assay.
RESULTS
Paclitaxel and NVP-BEZ235 cotreatment group can further inhibit the clonogenicity and invasion of Ishikawa and Ishikawa-TAX cells compared with paclitaxel alone and NVP-BEZ235 treatment group. Paclitaxel and NVP-BEZ235 cotreated groups increased the apoptosis rate of Ishikawa and increased G0/G1 phase arrest in both cells. Paclitaxel alone significantly inhibited p-AK and p-p70 S6K protein expression in Ishikawa and Ishikawa-TAX cells and the inhibition was enhanced by NVP-BEZ235 when cotreated with paclitaxel.
CONCLUSION
Paclitaxel can inhibit Ishikawa and Ishikawa-TAX cell via PI3K/Akt/mTOR signaling pathway. Paclitaxel and NVP-BEZ235 cotreatment can enhance the inhibitory effect.
目的
研究紫杉醇耐药型子宫内膜癌细胞的生物学特性及其耐药机制。
方法
采用高剂量紫杉醇（TAX）注射法建立紫杉醇耐药细胞系。用 CCK-8 法检测 Ishikawa 和 Ishikawa-TAX 细胞中紫杉醇的 IC50。用流式细胞术检测细胞周期和凋亡率。用 Western blot 检测 p-AKT 和 p-p70S6K 的表达。用 RT-PCR 检测耐药相关基因 Pgp 和 MDR1 的表达。用软琼脂克隆形成实验检测细胞活力，用 Transwell 体外侵袭实验检测细胞的侵袭能力。
结果
紫杉醇与 NVP-BEZ235 联合处理组比紫杉醇单药和 NVP-BEZ235 处理组能进一步抑制 Ishikawa 和 Ishikawa-TAX 细胞的集落形成和侵袭能力。紫杉醇与 NVP-BEZ235 联合处理组增加了 Ishikawa 细胞的凋亡率，并使两种细胞均发生 G0/G1 期阻滞。紫杉醇单药处理明显抑制了 Ishikawa 和 Ishikawa-TAX 细胞中 p-AKT 和 p-p70 S6K 蛋白的表达，而与 NVP-BEZ235 联合处理时增强了抑制作用。
结论
紫杉醇可通过 PI3K/Akt/mTOR 信号通路抑制 Ishikawa 和 Ishikawa-TAX 细胞。紫杉醇与 NVP-BEZ235 联合处理可增强抑制作用。
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