姜黄素调控子宫内膜癌细胞糖酵解过程的机制

目的：探究姜黄素靶向子宫内膜癌细胞糖酵解过程的调控机制，探索常规化疗药物联合姜黄素靶向杀伤子宫内膜癌细胞的作用效果。方法：培养子宫内膜癌Ishikawa细胞，利用能量代谢检测Seahorse、乳酸及丙酮酸水平，验证子宫内膜癌细胞糖酵解及氧化磷酸化水平；通过qPCR检测了糖酵解相关基因在姜黄素处理后子宫内膜癌细胞及正常子宫内膜癌细胞中表达情况，筛选姜黄素潜在的靶向调控位点；为了验证姜黄素通过丙酮酸脱氢酶激酶2（PDK2）调控子宫内膜癌细胞能量代谢过程，利用Seahorse及丙酮酸水平检测试实验，检测PDK2敲除后子宫内膜癌细胞能量代谢过程变化；小鼠体内皮下成瘤实验，探究姜黄素联合5-FU杀伤子宫内膜癌细胞的治疗效果。结果：姜黄素作用于子宫内膜癌细胞可显著抑制子宫内膜癌细胞糖酵解途径，而氧化磷酸化水平并未发生显著改变；姜黄素可显著下调PDK2的表达，调控子宫内膜癌糖酵解过程，而在子宫内膜癌细胞中敲除PDK2的表达，可显著抑制子宫内膜癌细胞的糖酵解途径，诱导子宫内膜细胞增殖抑制，因此姜黄素作用于PDK2调控子宫内膜癌糖酵解过程，从而达到杀伤肿瘤细胞的目的；5-FU联合姜黄素联合用药，可显著提高5-FU杀伤肿瘤细胞能力，抑制肿瘤细胞生长，5-FU联合姜黄素可显著提高子宫内膜癌细胞对化疗药物的敏感性。结论：姜黄素通过PDK2调控子宫内膜癌细胞糖酵解过程，从而抑制子宫内膜癌细胞增殖过程，增加子宫内膜癌细胞对5-FU的敏感性。

The mechanism of curcumin in regulating glycolysis of endometrial carcinoma cells

Objective: To detect the energy metabolic pathway of curcumin combined with 5-Fluorouridine and its killing effect on endometrial cancer cells. Methods: The levels of glycolysis and oxidative phosphorylation in endometrial cancer cells were measured by Seahores and Lactate/Pyruvate assay kit to confirm the main metabolic pathway in the endometrial cancer cells. Metabolic genes in Ishikawa and curcumin treated Ishikawa cells were screened by real-time PCR to explore the candidate target gene of curcumin; the level of glycolysis and oxidative phosphorylation were detected by Seahorse and the lactate/pyruvate assay in PDK2-knockout cells to reveal how curcumin regulated the energy metabolic pathway in endometrial carcinoma cells via PDK2. We establish human endometrial cancer model in nude mice to detect the treatment effect of 5-Fluoracil combined with curcumin in vivo. Results: It was revealed that curcumin could inhibit the glycolysis in the endometrial cancer cells, but the OXPHOS level couldn’t be changed. It was also found that curcumin could down-regulate the expression of pyruvate dehydrogenase kinase-2 (PDK2), which regulated the process of glycolysis. The knock-downed expression of PDK2 by specific shRNA decreased the level of glycolysis and proliferation of endometrial cancer cells. Curcumin could inhibit the level of glycolysis in the endometrial cancer cells by down-regulating the expression of PDK2. Combined with 5-FU, curcumin could obviously increase the sensitivity of endometrial cancer cells to chemotherapeutics, and up-regulate the ability to inhibit the proliferation of cancer cells. Conclusion: Curcumin could inhibit the expression of PDK2 and the level of glycolysis in the endometrial cancer cells.As a result, curcumin could obviously increase the sensitivity to 5-FU, and inhibit the proliferation of endometrial cancer cells effectively.