Study On The Mechanism Of Erzhi Pills In Invasion And Metastasis Of Colon Cancer Cells

[Objective]Invasion and metastasis is one of the important factors for the high mortality of colon cancer patients.This study explores the mechanism of Erzhi pills on the proliferation,cycle,apoptosis,invasion and metastasis of colon cancer cells based on the hypothesis of "yin deficiency causing tumor".It provides scientific basis for Erzhi Pills to resist proliferation,invasion and metastasis of colon cancer,and it is of great significance to improve the survival rate of the patients.[Methods]1.The experimental study of cell proliferation: MTT assay was used to detect the effect of erzhi pills containing serum and Oxaliplatin on cell proliferation2.The experimental study of cell cycle apoptosis: The cell cycle distribution was detected by PI staining;the cell apoptosis was detected by Annexin V-FITC/PI staining;detecting the expression of apoptosis-related proteins Bax and Bcl-2 by Western blot.3.The experimental study of cell invasion and metastasis: Detecting cell migration by scratch test;Detecting cell invasiveness by Transwell chamber;detecting the expression of p65?p-p65?VEGFR1 and E-Cadherin proteins by Western blot;detecting the expression of VEGFR1 and CDH1 mRNA by Western blot [Results]1.MTT experiment results show: After 12 hours treatment with Erzhi pills serum,2.5% and 10% drug serum significantly inhibited the proliferation of HCT116 cells(P<0.05).After 48 hours,2.5%,5%,10%,20% drug serum had inhibitory effect on cells(P<0.05).After 72 hours,only 10% drug serum had inhibitory effect on the cells(P<0.05).2.Cell cycle and apoptosis test:(1)Cycle test results show: compared with the blank control group,the cells in L-OHP group and combined group were arrested in G1 phase(P<0.05),the cells in medium and high Erzhi pills serum group were arrested in S phase(P<0.05).(2)Apoptosis test results show: compared with the blank control group,the apoptosis of the cells was significantly improved in each group(P<0.05),and the cell apoptosis rate was the highest in the combination group(P<0.05).(3)Western blot experiment results show: compared with the blank control group,the expression of Bax protein was significantly increased in the L-OHP group and the combination group(P<0.05),and the expression of Bax protein in the combination group was higher than that of the L-OHP group(P<0.05).Compared with the blank control group,the expression of Bcl-2 protein was significantly down-regulated in all the drug-treated groups(P<0.05).3.Invasion and metastasis test:(1)Scratch test results show: after 12 and 24 hours: compared with the blank control group,the cell migration rate of each drug group was lower(P<0.05),and the lowest in the combination group(P<0.05).after 36 and 48 hours: compared with the blank group,the cell migration rate decreased except for the high dose group of Erzhi pills(P<0.05),and the lowest in the combination group.(2)Transwell experiment results show: the number of perforating cells in each drug group was lower than blank control group(P<0.05).(3)Western blot experiment results show: compared with the blank control group,the medium and high dose group significantly reduced the expression of the p-p65 and VEGFR1 protein and the combination group was lower than L-OHP group(P<0.05).The expression of E-cadherin was significantly increased in the L-OHP group and the combination group compared with the blank control group and the combination group was higher than L-OHP group(P<0.05).(4)QPCR experiment results show: compared with the blank control group,the expression of VEGFR1 mRNA was significantly decreased in the medium and high dose group(P<0.05).The combination group was lower than L-OHP group(P<0.05).Compared with the blank control group,each group had no significant effect on the expression of CDH1 mRNA.[Conclusions]1.Erzhi Pills serum can inhibit the proliferation of Colon Cancer HCT116 cells.2.Erzhi Pills serum can change the cell cycle distribution of colon cancer HCT116 cells,block the cells in S phase,promote the apoptosis of cells,and strengthen the pro-apoptotic ability of L-OHP.The mechanism may be related to the inhibition of the expression of Bcl-2 protein and the increase of the expression of Bax protein.3.Erzhi Pills serum can inhibit the migration and invasion of colon cancer HCT116 cells,which may be related to inhibition of p-p65 protein,VEGFR1 protein and mRNA expression,and increase the expression of E-cadherin protein by synergistic L-OHP.