目的 探討臭椿酮對順鉑（DDP）耐藥胃癌細胞株SGC-7901/DDP耐藥性的影響及其機制。方法 采用噻唑藍（MTT）法檢測不同質(zhì)量濃度臭椿酮對SGC-7901/DDP細胞活力的影響以篩選無毒作用濃度。將體外培養(yǎng)的SGC-7901/DDP細胞分為臭椿酮低濃度（0.2 mg/mL）＋DDP（2 μg/mL）組、臭椿酮中濃度（0.4 mg/mL）＋DDP（2μg/mL）組、臭椿酮高濃度（0.8 mg/mL）＋DDP（2 μg/mL）組考察臭椿酮對SGC-7901/DDP細胞DDP敏感性的影響；將體外培養(yǎng)的SGC-7901/DDP細胞分為對照組、DDP（2 μg/mL）組、DDP（2 μg/mL）＋Pifithrin-α（20 μmol/L）組、臭椿酮（0.8 mg/mL）＋DDP組、臭椿酮＋DDP（2μg/mL）＋Pifithrin-α（20 μmol/L）組考察p53信號通路與藥物作用的關(guān)系；采用MTT法檢測SGC-7901/DDP細胞活力，流式細胞儀檢測SGC-7901/DDP細胞凋亡率，免疫印跡法檢測SGC-7901/DDP細胞中LC3Ⅱ、LC3Ⅰ、Beclin1和p53蛋白表達水平。結(jié)果 0.2、0.4、0.8 mg/mL臭椿酮對SGC-7901/DDP細胞未產(chǎn)生明顯細胞毒性。與DDP組相比，臭椿酮低濃度＋DDP組、臭椿酮中濃度＋DDP組、臭椿酮高濃度＋DDP組細胞活力明顯降低，凋亡率、LC3Ⅱ/LC3Ⅰ值和Beclin1、p53蛋白表達水平明顯升高（P<0.05），且呈濃度依賴性；而DDP組和對照組之間差異無統(tǒng)計學意義（P>0.05）；給予Pifithrin-α作用后的DDP＋Pifithrin-α組和臭椿酮＋DDP＋Pifithrin-α組細胞活力較相應對照DDP組、臭椿酮＋DDP組明顯升高，而細胞凋亡率、LC3Ⅱ/LC3Ⅰ值和Beclin1、p53蛋白表達水平較DDP組、臭椿酮＋DDP組明顯降低（P<0.05）。結(jié)論 臭椿酮可通過p53通路誘導自噬逆轉(zhuǎn)SGC-7901/DDP細胞對DDP的耐藥性。

Objective To investigate the effect of ailanthone (ATE) on the drug resistance of gastric cancer cell line SGC-7901/DDP to cisplatin and its mechanism. Methods Methyl thiazolyl tetrazolium (MTT) assay was used to detect the effects of different concentrations of ATE on SGC-7901/DDP cell viability to screen the non-toxic concentration. SGC-7901/DDP cells cultured in vitro were divided into low concentration ATE (0.2 mg/mL ATE) + DDP (2 μg/mL cisplatin), medium concentration ATE (0.4 mg/mL ATE) + DDP group, high concentration ATE (0.8 mg/mL ATE) + DDP group to investigate the effect of ailanthone on DDP sensitivity of SGC-7901/DDP cells. SGC-7901/DDP cells cultured in vitro were divided into control group, DDP group, DDP+ Pifithrin-α (20 μmol/L Pifithrin-α), ATE (0.8 mg/mL ATE)+DDP group, ATE+DDP+ Pifithrin-α group to investigate the relationship between p53 signaling pathway and drug action. MTT assay was used to detect the viability of SGC-7901/DDP cells, flow cytometry was used to detect the apoptosis rate of SGC-7901/DDP cells, Western blotting was used to detect the protein expression levels of LC3 Ⅱ, LC3 Ⅰ, Beclin1, and p53 in SGC-7901/DDP cells. Results 0.2, 0.4, and 0.8 mg/mL ATE had no cytotoxicity on SGC-7901/DDP cells. Compared with those in DDP group, the cell viability of low concentration ATE+DDP group, medium concentration ATE+DDP group and high concentration ATE+DDP group were significantly lower, the apoptosis rate, LC3 Ⅱ/LC3 Ⅰ value and Beclin1, p53 protein expression levels were significantly higher (P<0.05), and it was concentration dependent. There was no significant difference between DDP group and control group (P>0.05), after treatment with Pifithrin-α, the cell viability of DDP+ Pifithrin-α group and ATE+DDP+Pifithrin-α group was significantly higher than that of DDP group and ATE+DDP group, the apoptosis rate, LC3 Ⅱ/LC3 Ⅰ value and Beclin1, p53 protein expression levels were significantly lower (P<0.05). Conclusion ATE can induce autophagy and reverse cisplatin resistance of SGC-7901/DDP cells by p53 pathway.

R285.5

國家自然科學基金資助項目（81903770）