Impacts of Leonurine on the Proliferation, Apoptosis and Angiogenesis of Hepatoma SMC-7721 Cells by Regulating Shh/Gli1 Signaling Pathway
WANG Jian1, LÜ Xuemei2, QI Liting3*
The aim of this study was to investigate the impacts of Leo (Leonurine) on the proliferation, apoptosis and angiogenesis of hepatoma SMC-7721 cells by regulating Shh (Sonic hedgehog)/Gli1 (glioma associated oncogene homolog 1) signaling pathway. SMC-7721 cells were cultured in vitro and divided into: control group, low concentration Leo group (Leo-L group, 10 μg/mL Leo), medium concentration Leo group (Leo-M group, 20 μg/mL Leo), high concentration Leo group (Leo-H group, 30 μg/mL Leo), recombinant human SHH protein group (SHH group, 3 μg/mL recombinant human SHH protein), and high concentration Leo+recombinant human SHH protein group (Leo-H+SHH group, 30 μg/mL Leo+3 μg/mL recombinant human SHH protein). CCK8 experiment was applied to detect SMC-7721 cell proliferation. Scratch test was applied to detect SMC-7721 cell migration. Flow cytometry was applied to detect apoptosis in SMC-7721 cells. Lumen formation experiment was applied to detect angiogenesis in SMC-7721 cells. Western blot was applied to detect the Shh/Gli1 signaling pathway and apoptosis related protein expression levels in SMC-7721. Compared with the control group, the D450 value, scratch healing rate, angiogenesis quantity, and Bcl-2, Shh, and Gli1 protein expression of SMC-7721 cells in the Leo-L, Leo-M, and Leo-H groups were obviously reduced (P<0.05); the apoptosis rate and Bax protein expression were obviously increased (P<0.05); the D450 value, scratch healing rate, angiogenesis quantity, and Bcl-2, Shh, and Gli1 protein expression of SMC-7721 cells in the SHH group were obviously increased (P<0.05); the apoptosis rate and Bax protein expression were obviously reduced (P<0.05). Compared with the Leo-H group, the D450 value, scratch healing rate, angiogenesis quantity, and Bcl-2, Shh, and Gli1 protein expression of SMC-7721 cells in the Leo-H+SHH group were obviously increased (P<0.05); the apoptosis rate and Bax protein expression were obviously reduced (P<0.05). This suggests that Leo may inhibit the proliferation and angiogenesis of hepatoma SMC7721 cells and induce cell apoptosis by inhibiting the Shh/Gli1 signaling pathway.
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