The Mechanism Of 27-hydroxycholesterol On Glucose Metabolism In Human Lung Cancer A549 Cells

Objective : 27-Hydroxycholesterol(27HC or 27-OHC),the most abundant cholesterol primary metabolite in plasma,is an endogenous selective estrogen receptor modulator and LXR agonist.27 HC is closely related to the development of lung cancer,and 27 HC promotes lung cancer cell proliferation in an ER?-dependent manner.However,27 HC was found to be able to reduce intracellular cholesterol levels and inhibit cell proliferation by activating the LXR pathway in ER-negative A549 cells.Previous studies have shown that 27 HC plays an important role in lung cancer cell proliferation,invasion and migration,and cholesterol metabolism,but its research on glucose metabolism in A549 cells has not been reported yet.This study aimed to investigate the effects of 27 HC on glucose metabolism in A549 cells.The expression of LXR,GLUT1,HK2,PKM2,LDHA and PCK2 in A549 cells treated with 27 HC,T0901317 or si LXR?,ATP levels,glucose uptake and lactate production were examined,and the LXR signaling pathway was further explored in this process.The possible mechanism of action to further discover the potential role of 27 HC in the pathogenesis and prognosis of cancer.Methods1.A549 cells were treated with different concentrations(0~15?M)27HC and(0~25?M)T0901317 for 12~48 hours.2.2.CCK8 kit detects the viability of A549 cells.3.si LXR? interferes with the expression of LXR? in A549 cells.4.ATP test kit detects intracellular ATP levels.5.The glucose test kit and the lactic acid test kit respectively detect the contents of glucose and lactic acid in the culture solution.6.Western blot was used to detect the expression of LXR? and glucose metabolism related proteins GLUT1,HK2,PKM2,LDHA and PCK2.Results1.27 HC concentration(1~15?M,24h)and time(0~48h,5?M)depended on inhibiting the viability of A549 cells;2.The concentration of T0901317(1~25?M,24h)inhibited the viability of A549 cells;3.27 HC inhibited glucose metabolism in A549 cells: 27 HC concentration(1~15?M,24h)and time(0~48h,5?M)depended on activation of LXR?,decreased ATP level of A549 cells,decreased glucose uptake and lactate production,Inhibition of expression of GLUT1,HK2,PKM2,LDHA and PCK2 proteins;4.T0901317 inhibits glucose metabolism in A549 cells,and is consistent with the effect of 27HC: T0901317 concentration(5?M,24h)activates LXR?,decreases ATP levels in A549 cells,reduces glucose uptake and lactate production,and inhibits GLUT1,HK2,PKM2,LDHAAnd expression of PCK2 protein;5.Knockdown of LXR? attenuated the inhibitory effects of 27 HC and T0901317 on glucose metabolism in A549 cells: Compared with the27 HC or T0901317-treated group,A549 cells in the si LXR? group had elevated ATP levels,increased glucose uptake and lactate secretion,and GLUT1,HK2,PKM2,LDHA and PCK2 protein expression is upregulated.Conclusions:1.27 HC inhibits glucose metabolism of A549 cells;2.27 HC participates in the glucose metabolism process of A549 cells through LXR signaling pathway.