飞燕草素对三阴性乳腺癌MDA-MB-231细胞糖酵解的影响

目的:探讨飞燕草素对乳腺癌MDA-MB-231细胞糖代谢的影响。方法:以不同浓度的飞燕草素作用MDA-MB-231细胞,CCK-8检测细胞活性,免疫组化染色检测Ki67表达,划痕试验检测细胞的迁移能力;葡萄糖摄取、乳糖和三磷酸腺苷(ATP)测定试剂盒检测MDA-MB-231细胞葡萄糖摄取、乳糖和ATP的生成,己糖激酶(HK)活性试剂盒检测HK活性;蛋白质印迹法检测HK、丙酮酸激酶(PK)、乳酸脱氢酶A(LDHA)和M2-型丙酮酸激酶 2(PKM2)表达。结果:飞燕草素能明显抑制MDA-MB-231细胞增殖,20和40 μmol/L飞燕草素能分别降低细胞存活率至75%和72%(P<0.05);20、40 μmol/L飞燕草素能降低乳腺癌MDA-MB-231细胞Ki67表达阳性的细胞数量,差异有统计学意义(P<0.01);飞燕草素能降低乳腺癌细胞葡萄糖的摄取,降低细胞乳糖和ATP的生成,抑制PKM2、HK和LDHA蛋白的表达,降低p-Akt和p-mTOR蛋白表达。结论:飞燕草素能抑制乳腺癌MDA-MB-231细胞糖代谢,抑制Akt/mTOR通路活性可能是飞燕草素抑制乳腺癌细胞糖酵解的机制。

Effect of Delphinidin on Glycolysis in Triple-Negative Breast Cancer Cell MDA-MB-231

Objective:To research the effect of delphinidin on glycolysis in breast cancer cell MDA-MB-231.Methods:Cell counting kit-8(CCK8)assay and Ki67 immunohistochemistry were used to test the effect of delphinidin on breast cancer cell MDA-MB-231 proliferation,wound healing assay was used to test the effect of delphinidin on MDA-MB-231cells migration. The effect of delphinidin on glucose uptake,lactate and ATP generation and the activity of hexokinase(HK)were detected in MDA-MB-231cells. Protein expression of pyruvatekinase type M2 pyruvatekinase(PKM2),HK,lactate dehydrogenase-A(LDHA),p-Akt and p-mTOR were detected with western blot(WB). Results:Delphinidin inhibited MDA-MB-231 cells proliferation(Ki67 positive cells)and cell viability. 20 and 40 μmol/L delphinium could reduce the cell survival rate to 75% and 72% respectively(P<0.05). 20 and 40 μmol/L delphinium could reduce the number of Ki67-positive breast cancer MDA-MB-231 cells,the difference was statistically significant(P<0.01). Delphinidin also inhibited glucose uptake,decreased lactate and ATP generation in MDA-MB-231 cells. WB results showed that delphinidin inhibited the protein expression of PKM2,HK,LDHA,p-Akt and p-mTOR. Conclusion:The delphinidin could inhibit glucose uptake and glycolysis in MDA-MB-231 cells. Delphinidin might inhibit MDA-MB-231 cells glycolysis through Akt/mTOR pathway.