目的 探讨马齿苋酰胺E对人肾癌786-O细胞增殖、迁移和侵袭的作用及机制。方法 体外培养786-O细胞，给予马齿苋酰胺E （10、20、40 μmol/L）进行干预，采用CCK-8法检测细胞增殖情况；采用划痕实验检测细胞迁移能力；采用Transwell小室法检测细胞侵袭能力；采用AO/EB试剂盒检测细胞凋亡率。建立裸鼠移植瘤肾癌模型，给予马齿苋酰胺E （3、15 mg/kg）进行干预，测定各组裸鼠肿瘤质量及体积，采用苏木素-伊红（HE）染色法观察肿瘤组织病理变化；采用Western blotting法检测786-O细胞和荷瘤裸鼠瘤组织中细胞增殖标志物如Ki67、增殖细胞核抗原（proliferating cell nuclear antigen，PCNA）及凋亡标志物如活化半胱氨酸蛋白酶-3（Cleaved Caspase-3）、Cleaved Caspase-9及侵袭相关蛋白如基质金属蛋白酶-2（matrix metalloproteinase-2，MMP-2）、MMP-9及希佩尔-林道抑癌基因（Von Hippel-Lindau，VHL）/缺氧诱导因子-1α（hypoxia inducible factor-1α，HIF-1α）通路相关蛋白VHL、HIF-1α的表达情况。结果 马齿苋酰胺E显著抑制786-O细胞活力（P<0.05、0.01），下调Ki67、PCNA蛋白表达水平（P<0.05、0.01）；明显促进786-O细胞凋亡（P<0.05、0.01），上调Cleaved Caspase-3、Cleaved Caspase-9蛋白表达水平（P<0.05、0.01）；有效改善786-O细胞迁移与侵袭能力（P<0.05、0.01），显著降低MMP-2、MMP-9蛋白表达水平（P<0.05、0.01）；显著上调VHL蛋白表达水平（P<0.05、0.01）并显著下调HIF-1α蛋白表达水平（P<0.01）。马齿苋酰胺E可有效降低荷瘤裸鼠肿瘤质量与体积（P<0.05、0.01），抑制肿瘤细胞增殖并促进其凋亡，显著下调Ki-67、PCNA、MMP-2、MMP-9、HIF-1α蛋白表达水平（P<0.05、0.01），显著上调Cleaved Caspase-3、Cleaved Caspase-9、VHL蛋白表达水平（P<0.05、0.01）。结论 马齿苋酰胺E体外可有效抑制786-O细胞增殖、迁移与侵袭，促进786-O细胞凋亡，体内可有效降低荷瘤裸鼠肿瘤质量与体积，其作用机制可能与调控VHL/HIF通路及其下游蛋白表达有关。

Objective To investigate the effect and mechanism of oleracein E on proliferation, invasion, and migration of human renal carcinoma 786-O cells. Methods Human renal carcinoma 786-O cells were cultured in vitro and treated with oleracein E (10, 20, and 40 μmol/L) for intervention. CCK-8 method was used to detect cell proliferation; Scratch test was used to detect cell migration ability; Transwell chamber method was used detect cell invasion ability; AO/EB kit was used to detect cell apoptosis rate. Kidney cancer model of transplanted tumor in nude mice was established and treated with oleracein E (3, 15 mg/kg) for intervention. Weight and volume of tumor in nude mice was determined, pathology changes of tumor tissue were observed by hematoxylin-eosin (HE) staining. Western blotting was used to detect expressions of cell proliferation marker such as Ki67 and proliferating cell nuclear antigen (PCNA), apoptosis marker such as activated cysteine protease-3 (Cleaved Caspase-3) and Cleaved Caspase-9, invasion-related protein such as matrix metalloproteinase-2 (MMP-2) and MMP-9, Von Hippel-Lindau (VHL)/hypoxia inducible factor-1α (HIF-1α) pathway related proteins such as VHL and HIF-1α in 786-O cells and tumor tissues of nude mice. Results Viability of 786-O cells was significantly inhibited (P < 0.05, 0.01), expressions of Ki-67 and PCNA were down-regulated (P < 0.05, 0.01); Apoptosis of 786-O cells was significantly promoted (P < 0.05, 0.01), expressions of Cleaved Caspase-3 and Cleaved Caspase-9 were up-regulated (P < 0.05, 0.01); Migration and invasion ability of 786-O cells were effectively improved (P < 0.05, 0.01), expressions of MMP-2 and MMP-9 were significantly reduced (P < 0.05, 0.01); VHL protein expression was significantly up-regulated (P < 0.05, 0.01) and HIF-1α protein expression was significantly down-regulated (P < 0.01) by oleracein E. Weight and volume of tumor in tumor-bearing nude mice were effectively reduced (P < 0.05, 0.01), proliferation of tumor cells was inhibited and apoptosis of tumor cells was promoted, Ki67, PCNA, MMP-2, MMP-9, HIF-1α expression were significantly down-regulated (P < 0.05, 0.01), expressions of Cleaved Caspase-3, Cleaved Caspase-9, and VHL were significantly up-regulated (P < 0.05, 0.01) by oleracein E. Conclusion Oeracein E can effectively inhibit the proliferation, migration, and invasion of 786-O cells in vitro, promote the apoptosis of 786-O cells, and effectively reduce the tumor weight and volume of tumor-bearing nude mice in vivo, of which mechanism may be related to the regulation of VHL/HIF pathway and its downstream protein expression.

R285.5

浙江省教育厅科研项目（Y202045313）