| 王 俊,王 聪,徐超亮,鲁 军,邵 怡.敲低PFKP联合肉碱棕榈酰基转移酶抑制剂etomoxir协同对肾透明细胞癌细胞Caki-1的抗肿瘤作用[J].,2019,19(23):4411-4414 |
| 敲低PFKP联合肉碱棕榈酰基转移酶抑制剂etomoxir协同对肾透明细胞癌细胞Caki-1的抗肿瘤作用 |
| The Anti-tumor Effect of PFKP Knockdown Combined with Carnitine Palmitoyltransferase Inhibitor Etomoxir on Renal Clear Cell Carcinoma Cell Line Caki-1 |
| 投稿时间：2019-04-28 修订日期：2019-05-23 |
| DOI：10.13241/j.cnki.pmb.2019.23.003 |
| 中文关键词: PFKP 肾透明细胞癌 细胞死亡 肿瘤代谢 |
| 英文关键词: PFKP Renal clear cell carcinoma Cell death Cancer metabolism |
| 基金项目:上海交通大学"医工交叉基金"项目（YG2016MS20） |
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| 中文摘要: |
| 摘要 目的：研究敲低P型磷酸果糖激酶（phosphofructokinase，PFKP）联合肉碱棕榈酰基转移酶抑制剂etomoxir对肾透明细胞癌Caki-1细胞的影响，并进一步探究其作用机制。方法：利用Western blot验证对照及PFKP shRNA敲低肾透明细胞癌细胞中 PFKP的敲低效率，分别检测对照组（shCtrl）、PFKP敲低组、etomoxir组（shCtrl+etomoxir）、PFKP敲低联合etomoxir组的增殖曲线。使用Annexin-V/PI染色并用流式细胞检测对照组、PFKP敲低组、etomoxir组、PFKP敲低联合etomoxir组的细胞死亡，研究 PFKP敲低联合etomoxir对细胞存活的影响。分别检测对照组、PFKP敲低组、etomoxir组、PFKP敲低联合etomoxir组的ATP水平与脂肪酸变化。结果：Western blot结果验证了PFKP的敲低效率。流式细胞检测显示，对照组、PFKP敲低组、etomoxir组、PFKP敲低联合etomoxir组的平均细胞死亡率分别为1.1、1.9、13.9、31.3%。PFKP敲低联合etomoxir组Caki-1细胞的死亡率显著高于单纯PFKP敲低与etomoxir组（P<0.05）。PFKP敲低联合etomoxir组Caki-1细胞的ATP水平显著低于单纯PFKP敲低与etomoxir组（P<0.05）。Etomoxir的加入抑制了PFKP敲低引起的游离脂肪酸下降（P<0.05）。结论：PFKP敲低联合etomoxir对Caki-1细胞呈现协同的细胞毒抗肿瘤作用。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the effect of P-type phosphofructokinase (PFKP) knockdown combined with carnitine palmitoyltransferase inhibitor etomoxir on the renal clear cell carcinoma Caki-1 cells, and further explore its mechanism of action. Methods: The knockdown efficiency of PFKP in renal clear cell carcinoma cells was confirmed by Western blot. The proliferation curves of control group (shCtrl), PFKP knockdown group, etomoxir group (shCtrl + etomoxir) and PFKP knockdown combined with etomoxir group were measured. The effects on cell survival in all groups were detected by flow cytometry using Annevin-V/PI staining. The ATP levels and free fatty acid levels in the control group, PFKP knockdown group, etomoxir group, PFKP knockdown combined with etomoxir group were detected. Results: Western blot analysis confirmed the knockdown efficiency of PFKP. Flow cytometry showed that the mean cell mortality rate of control group, PFKP knockdown group, etomoxir group, PFKP knockdown combined with etomoxir group were 1.1, 1.9, 13.9, 31.3% Compared with the control cells, the cell mortality of PFKP knockdown combined with etomoxir group Caki-1 cells was significantly higher than that of PFKP knockdown and etomoxir groups(P<0.05). The ATP levels in PFKP knockdown combined with etomoxir group were significantly lower than those of PFKP knockdown and etomoxir groups (P<0.05). The addition of etomoxir inhibited the decrease in free fatty acids caused by PFKP knockdown (P<0.05). Conclusion: PFKP knockdown combined with etomoxir showed synergistic cytotoxic and anti-tumor effects on Caki-1 cells. |
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