Mechanism Of Nerve Growth Factor Inhibiting Proliferation Of Hepatoma Cells By PTEN Gene

In this paper,Sanger sequencing was used to test the mutation of PTEN gene in hepatocellular carcinoma cell lines HepG2 and HuH7.The results showed that the wild-type PTEN gene sequence was included in HepG2 cells,and the PTEN gene mutation was detected in HuH7 cells.Therefore,the hepatoma cell line HepG2 was selected for subsequent experiments.Western Blotting was used to detect the expression level of PTEN protein in HepG2 cells and alpha-fetoprotein AFP in liver cancer cells.The results showed that the protein expression levels were negatively correlated.NGF is the earliest study and the most well-known neurotrophic factor.In recent years,experimental studies have shown that NGF plays an important role in the occurrence and development of tumors during tumor growth,differentiation,angiogenesis and metastasis.In order to further explore the mechanism of NGF in hepatocarcinoma cells,this study used NGF to treat wild-type hepatoma cells HepG2 and PTEN knockout HepG2 cells to detect cell proliferation,apoptosis and cycle changes.The results showed that NGF can effectively inhibit the proliferation of wild-type HepG2 cells,promote their apoptosis and induce intracellular G0/G1 cycle arrest.For PTEN knockout HepG2 cells,NGF has no significant proliferation,apoptosis and cycle.In order to further explore the negative correlation between PTEN protein and AFP protein in hepatocarcinoma cells,HepG2 cells were treated with EGF,ATRA and NGF respectively.The results of qRT-PCR and Western Blotting showed that EGF did not affect the expression of PTEN and AFP in HepG2 cells;NGF and ATRA can significantly alter the expression of PTEN and AFP.On this basis,the PTEN gene knockout HepG2 cells were obtained by CRISPR/Cas9 gene editing technology,and NGF,PI3K inhibitor Wortmannin and Akt inhibitor Perifosine,qRT-PCR and Western Blotting results were used to simultaneously treat HepG2 with NGF and Wortmannin.The cells did not change the expression of PTEN and AFP in HepG2.Simultaneous addition of NGF and Perifosine only changed the expression of PTEN in HepG2 cells,and did not change the expression of AFP.The expression of AFP was not significantly changed by HepG2 cells treated with NGF.In this study,we explored the function and mechanism of PTEN gene in hepatocarcinoma cells,and found that NGF promoted the expression of PTEN in HepG2 cells,inhibited the expression of AFP in HepG2,and revealed that the expression level was related to PI3K/Akt cell pathway.This finding provides a new and exploreable path for clinical treatment of liver cancer.