Study on the effects of active components of Lichong Shengsui decoction on autophagy in ovarian cancer by regulating related signaling pathways

Liu Fangyuan; Ding Danni; Guo Ying; Shen Ying; Li Jia; Han Fengjuan

doi:10.3760/j.issn.1673-4777.2024.05.004

Abstract：Objective To observe the effect of Lichong Shengsui decoction(LCSSY)on autophagy in ovarian cancer by regulating adenosine monophosphate-activated protein kinase(AMPK)/mammalian target of rapamycin(mTOR)/uncoordinated 51 like kinase-1(ULK1)signaling pathway.Methods Ovarian cancer SKOV3 cells were cultured in vitro and serum containing LCSSY was prepared.SD rats were divided into blank control group and LCSSY low,medium and high dose groups by random number table method.Cell count kit-8 method was used to detect the proliferation inhibition rate of ovarian cancer SKOV3 cells.Western blotting was used to detect the expression of autophagy related proteins recombinant human autophagy effector protein(Beclin-1),ratio of human microtubule associated protein light chain 3 Ⅱ to mouse microtubule associated protein light chain 3 Ⅰ(LC3Ⅱ/Ⅰ),autophagy junction protein(P62),as well as AMPK/mTOR/ULK1 pathway related proteins.AMPK activator was used to verify its mechanism of action.Results At 24 and 48 h after administration,the inhibition rates of SKOV3 cell proliferation in the low,medium and high dose LCSSY groups were higher than those in the blank control group,and the inhibition degree was gradually strengthened with the increase of drug concentration(all P＜0.05).In addition,the inhibition rates of low dose LCSSY group and medium dose LCSSY group at 48 h were higher than those at 24 h(both P＜0.05).Compared with the blank control group,the expression of Beclin-1 and LC3 Ⅱ/Ⅰ was down-regulated in the low,medium,and high dose LCSSY groups in a dose-dependent manner[(0.98±0.02),(0.59±0.02),(0.55±0.02)vs(1.46±0.12);(4.45±0.21),(4.00±0.25),(2.95±0.21)vs(5.98±0.23);(0.67±0.02),(0.94±0.05),(1.53±0.13)vs(0.30±0.02)](all P＜0.05).Compared with the blank control group,the expression of AMPK and ULK1 was decreased,and the expression of mTOR complex 1 was increased in low,medium and high dose LCSSY groups in a dose-dependent manner(all P＜0.05).AMPK activator reversed the AMPK/mTOR/ULK1 signaling pathway,and compared with the high dose LCSSY group,the high dose of LCSSY combined with AMPK activator increased the protein expressions of AMPK and ULK1,while the protein expression of mTOR complex 1 decreased(all P＜0.05).Conclusion LCSSY may inhibit autophagy in ovarian cancer cells by regulating AMPK/mTOR/ULK1 signaling pathway.