秦皮甲素抑制人鼻咽癌细胞系HNE-3增殖

Esculin inhibits the proliferation of human nasopharyngeal carcinoma cell line HNE-3

目的 探讨秦皮甲素对鼻咽癌细胞生物学行为的影响及其可能作用机制。方法 不同浓度的秦皮甲素处理人鼻咽癌细胞系HNE-3;并将si-NC、si-circ-ZNF609转染至HNE-3细胞,pcDNA、pcDNA-circ-ZNF609分别转染至HNE-3细胞后用秦皮甲素处理细胞;采用CCK-8法检测细胞增殖;采用试剂盒检测细胞内葡萄糖摄取水平与乳酸水平;采用酶活性试剂盒检测丙酮酸激酶和己糖激酶的活性;Transwell小室法检测细胞迁移及侵袭;RT-qPCR检测circ-ZNF609的表达量;Western blot检测MMP-2、MMP-9蛋白表达量。结果 秦皮甲素以浓度依赖性的方式降低细胞存活率和MMP-2、MMP-9蛋白水平(P<0.001),降低葡萄糖摄取水平和乳酸水平(P<0.001),降低丙酮酸激酶和己糖激酶的活性(P<0.001),减少迁移及侵袭细胞数(P<0.001);秦皮甲素可降低circ-ZNF609的表达量(P<0.001),且呈剂量依赖性;转染si-circ-ZNF609后细胞存活率和MMP-2、MMP-9蛋白水平降低(P<0.001),葡萄糖摄取水平和乳酸水平降低(P<0.001),丙酮酸激酶和己糖激酶的活性降低(P<0.001),迁移及侵袭细胞数减少(P<0.001);转染pcDNA-circ-ZNF609能够回复秦皮甲素对HNE-3细胞增殖、糖酵解、迁移及侵袭的抑制作用。结论 秦皮甲素可通过调控circ-ZNF609表达而抑制HNE-3细胞增殖、糖酵解、迁移及侵袭。

Objective To explore the effect of esculin on the biological behavior of nasopharyngeal carcinoma cells and its possible mechanism. Methods Different concentrations of esculin were used to treat human nasopharyngeal carcinoma cells HNE-3, si-NC, si-circ-ZNF609 were transfected into HNE-3 cells, pcDNA, pcDNA-circ-ZNF609 were respectively transfected into HNE-3 cells and then treated with esculin. CCK-8 experiment was used to detect cell proliferation. Intracellular glucose uptake levels and lactate levels were tested using kits. The activity of pyruvate kinase and hexokinase was tested using an enzyme activity kit. The Transwell chamber test was used to find cell migration and invasion. RT-qPCR method was used to detect the expression of circ-ZNF609. Protein expression of MMP-2 and MMP-9 was calculated by Western blot. Results Esculin could reduce cell survival rate and theprotein levels of MMP-2, MMP-9 in a concentration-dependent manner(P<0.001), could reduce glucose uptake and lactate levels(P<0.001), and may reduce pyruvate kinase and the activity of hexose kinase(P<0.001), could also reduce the number of migrating and invading cells(P<0.001). Esculin could reduce the expression of circ-ZNF609(P<0.001), and it was dose-dependent. After transfection with si-circ-ZNF609, the cell survival rate and the protein levels of MMP-2, MMP-9 were decreased(P<0.001), glucose uptake and lactate levels were decreased(P<0.001). The activity of pyruvate kinase and hexokinase and the number of migrating and invasive cells were all decreased(P<0.001). Transfection of pcDNA-circ-ZNF609 may restore the inhibitory effects of esculin on the proliferation, glycolysis, migration and invasion of HNE-3 cells. Conclusions Esculin could inhibit cell proliferation, glycolysis, migration and invasion in HNE-3 cells by regulating the expression of circ-ZNF609.