Inhibitory Effect Of Apigetrin On Hela Cells Of Human Cervical Cancer And Its Molecular Mechanism

Cervical cancer is women’s mortality rate in the first four of cancer,and there is a greatly risk of high recurrence rate and side effects about its current treatment.Epidemiological studies have proved that flavonoids in several types of cancer(especially women)have the inhibition.Apigetrin,a kind of flavonoids,has an anti-cancer activity.At present,due to the characteristics of low toxicity and safety of natural active substances,more and more scientists have been used as dietary supplement medication-assisted therapeutic agents for research.In this study,transcriptome and in vitro cell experiments were used to study the inhibitory mechanism of on cervical cancer Hela cells.The main research contents and results are as follows(1)MTT experiment showed that apigetrin can inhibit the proliferation of Hela cells and the half-inhibitory concentration was 47.26 μM.Hela cells were treated with a concentration of 40 μM apigetrin,and after transcriptome processing,the expression levels of 7359 genes in Hela cells changed.Among them,the expression levels of 2873 genes increased and 4486 genes decreased.We mainly screened three gene enrichment pathways: the internal plasma reticulum protein processing signaling pathways,cell cycle pathways,and autophagy-animal pathways.q PCR was used to verify some significantly different genes,and the results were consistent with the expression profile.(2)The pro-apoptotic effect and molecular mechanism of apigetrin on cervical cancer Hela cells.It was determined by flow cytometry that apigetrin can promote Hela cell apoptosis in a dose-dependent manner,block the cell cycle in the G0/G1 phase,and reduce the membrane potential of mitochondria.q PCR and WB were used to detected the specific mechanism about anticancer of apigetrin.The results showed that it can regulates cycle-related genes and proteins such as CDK2/6,Cyclin A/D/E,and p16(INK4A),death receptor pathway,and mitochondrial pathway-related genes and proteins such as Fas L,Fas,TNF-R1,Bak,Bax,Bcl-2,Bad,Bcl-xl,cytochrome c,and caspase 3/8/9 to promote cell apoptosis.Besides,the PTEN/PI3K/AKT signaling pathway is also involved in Hela cell apoptosis induced by apigetrin.(3)The effect and molecular mechanism of apigetrin on promoting autophagy and migration of cervical cancer Hela cells.Apigetrin can enhance the autophagy fluorescence of LC3 B,combining q PCR and WB experiments to study autophagy-related genes and proteins: LC3 B and the addition of autophagy inhibitor 3-MA indicate that autophagy has a certain relationship with ROS and apoptosis;It also can affect the autophagy of Hela cells by regulating endoplasmic reticulum pathway-related genes and proteins ATF6 and chop;the trend of migration-related genes and protein MMP2/9 is consistent with the results of cell scratch healing experiments and migration experiments,proving that apigetrin can inhibit migration and invasion of Hela cells in cervical cancer.