Arisaema heterophyllum Blume Monomer Stigmasterol Targets PPARγ and Inhibits the Viability and Tumorigenicity of Lung Adenocarcinoma Cells NCI-H1975

Abstract

To clarify the regulatory effect and molecular mechanism of Arisaema heterophyllum Blume (AhBl) monomer stigmasterol on lung adenocarcinoma in human lung adenocarcinoma cells NCI-H1975 cultured in vitro and in nude mice. Oil red O staining, free fatty acid detection, adenosine triphosphate (ATP), and NADPH were applied to elucidate the regulatory effect of stigmasterol on the energy metabolism of NCI-H1975 cells. Simultaneously, colony formation assay and nude mouse tumorigenesis were performed to clarify the underlying mechanisms of stigmasterol on the proliferation and tumorigenesis of NCI-H1975 cells. Furthermore, peroxisome proliferator-activated receptor gamma (PPARγ) inhibitor GW9662 was supplemented to determine the expression changes of cyclins to clarify the regulation mechanism of stigmasterol. The results revealed that stigmasterol administration markedly inhibited the viability but promoted lipid deposition of NCI-H1975 cells. Meanwhile, the reduction of cell energy metabolism affected cell proliferation and colony formation. qPCR and western blot assays indicated that stigmasterol played a role in regulating the expression of cyclins and PPARγ signaling pathway proteins. Nude mouse tumorigenesis suggested that tumor size and weight in the stigmasterol-treated group were apparently lower as compared with the control group. Tumor tissue cells developed varying degrees of degeneration and large areas of ischemic necrosis presented in the central and peripheral cells. Immunohistochemistry results revealed that Ki67 expression in the stigmasterol group was substantially inhibited, while PPARγ expression was greatly elevated as compared with the control. GW9662 could mediate the inhibitory effect of stigmasterol on NCI-H1975 cells. The current study demonstrated that stigmasterol targeted PPARγ and inhibited the viability and tumorigenicity of lung adenocarcinoma cells NCI-H1975.

Advances in Stigmasterol on its anti-tumor effect and mechanism of action

“…The study of Song et al. ( 24 ) performed in vivo and in vitro experiments to investigate the regulatory role of stigmasterol in LUAD and try to clarify the corresponding molecular mechanism of action. They found that stigmasterol distinctly inhibited the viability of NCI-H1975 cells but promoted lipid deposition.…”

“…Studies have unraveled various biological and pharmaceutical properties of stigmasterol, such as analgesia ( 8 ), anti-inflammation ( 9 – 14 ), anti-oxidization ( 15 , 16 ), anti-diabetes ( 15 , 17 , 18 ), maintaining psychiatric status ( 19 ), lowering blood cholesterol level ( 20 , 21 ), improving learning and memory ability ( 22 ), and protecting against Leishmania ( 6 ), etc. Moreover, stigmasterol is recently reported with anti-tumor potential either in vivo or in vitro in several cancers (e.g., lung cancer ( 23 , 24 ), liver cancer ( 25 – 27 ), gallbladder cancer ( 28 , 29 ), gastric cancer ( 30 , 31 ), and ovarian cancer ( 32 )) via inhibiting growth while promoting apoptosis in tumor cells ( Figure 1 ).…”