The Effect And Mechanism Of Histone MONO-ADP-Ribosylation On HUVECs Proliferation And Migration In Human Colorectal Cancer LOVO Cell Lines

Objective: To investigate the effect and possible mechanism of proliferation and migration of HUVECs regulated by histone mono-ADP-ribosylation of LOVO cells of human colorectal cancer.Method: 1. Lentivirus was constructed through mutation at the locus of mono-ADP-ribosylation of histone from arginine to alanine or lysine transfected into LOVO cells. The stably transfected LOVO cell lines were selected as experimental group. The untransfected LOVO cells and those transfected with lentiviral vector were served as control group. 2. Supernatant of cultured LOVO cell was added to nutrient solutions of HUVECs, the effect on proliferation of HUVECs was then detected by CCK-8. Meanwhile, the migration ability of HUVECs was detected by Transwell. 3. mRNA expression of PTEN was detected by qRT-PCR, while expressions of H3K4me3、PTEN、PI3K、Akt、p-Akt、HIF-1α and VEGF were detected by western blotting.Results: 1. The results of transwell showed that the migration numbers of HUVECs were significantly reduced in experiment groups as compared to those in control groups(P<0.05). The inhibition rate of migration was then calculated to be 74.6%. The results of CCK-8 showed that the proliferation of HUVECs were also significantly reduced in experiment groups as compared to those in control groups(P<0.05). And the inhibition rate of proliferation decreases as the concentration of culture supernatant of LOVO cells decreases. 2. qRT-PCR showed that the mRNA level of PTEN was upregulated. The expressions of H3K4me3 and PTEN were upregulated, while PI3 K, p-Akt, HIF-1α and VEGF were downregulated in western blotting.Conclusions: 1. The ability of migration and proliferation was significantly inhibited after the mono-ADP-ribosylation of histone in LOVO cells reduced, which indicates that mono-ADP-ribosylation of histone may be involved in the regulation of migration and proliferation of HUVECs.2. Reduced mono-ADP-ribosylation of histone in LOVO cells could upregulate the expression of H3K4me3 and PTEN, meanwhile downregulate the expression of PI3K、p-Akt、HIF-1α and VEGF, which indicates that the PI3K/Akt signal pathway was inhibited when PTEN was activated. The downstream factors as HIF-1α and VEGF were also inhibited which leading to the inhibition of proliferation and migration of HUVECs.