The Antitumor Mechanism Of PAHDAA And Its Copper Complex In Hepatocellular Carcinoma Cells

BackgroundPrimary liver cancer is clinical common malignant tumors,the incidence of liver cancer worldwide,each year more than 626 thousand people,and there is an increasing trend in the incidence of malignant tumors,and ranksthe mortality of fifth,nearly 600 thousand / year,malignant tumor related death third.China is a large country with primary liver cancer,the number of patients accounted for about 55% of the total number of patients,liver cancer mortality in China ranked among the top second tumors.In recent years,although thegreat progress has been made in the early diagnosis and treatment measures,such as treatment based on surgery and the survival rate of patients with hepatocellular carcinoma has been improved,but the prognosis is still poor,the leading cause of liver cancer surgery is liver cancer recurrence and poor prognosis of intrahepatic and extrahepatic metastasis;However,research shows that liver cancer the occurrence,degradation,invasion and metastasis and angiogenesis are closely related,thus inhibiting tumor angiogenesis is one of the important means of the current anti-tumor therapy.ObjectiveIn order to further research and development of low toxicity and efficient anti-tumor angiogenesis drugs,this article will this laboratory synthetic drugs PAHDAA and PAHDAA-Cu anti-tumor angiogenesis research,preliminary expounds their antitumor mechanism of invasion and metastasis.Methods1.The structure of PAHDAA and PAHDAA-Cu and the complexation ratio of PAHDAA and Cu2+ were detected.2.The inhibitory effect of PAHDAA and PAHDAA-Cu on the growth of hepatocellular carcinoma HepG2 cells and HUVEC cells was detected by succinate dehydrogenase(MTT)assay.3.The ability of PAHDAA and PAHDAA-Cu to produce ROS in and out of cells by fluorescence photometer.4.RT-PCR and Western Blot were used to detect the invasion and metastasis of PAHDAA and PAHDAA-Cu cells,respectively.5.The effect of PAHDAA and PAHDAA-Cu on cell migration was detected by scratch test and Transwell assay.6.Through the observation of the chick chorioallantoic membrane assay effect of PAHDAA and PAHDAA-Cu on neovascularization.Results1.The PAHDAA and PAHDAA-Cu prepared in laboratory were soluble in 70% of methyl sulfoxide,and the complexation ratio of PAHDAA to copper was(2:1).2.PAHDAA and PAHDAA-Cu can inhibit the proliferation of hepatocellular carcinoma HepG2 cells and HUVEC cells(HepG2 cells,IC50=23.045±0.014 mol/L;HUVEC cells: IC50=53.942±0.014 mol/L),thePAHDAA-Cu showed growth inhibition of tumor cells,(HepG2 cells IC50=33.087±0.009: mol/L;HUVEC: IC50=26.245±0.013 mol/L cells).3.Fluorescence detection and reactive oxygen species assay showed that PAHDAA and PAHDAA-Cu could reduce the formation of free radicals in cells.4.RT-PCR and Western Blot results showed that the resistance to invasion and metastasis of tumor protein P53 activation and NDRG1 expression and E-cadherin expression were increased,the expression of HIF and downregulation of P65 protein showed good Kang tumor invasion and metastasis;no obvious changes in the expression of VEGF protein.5.HUVEC cell scratch test and Transwell test showed that PAHDAA and PAHDAA-Cu had stronger inhibitory effect on cell migration,and lattertheir complexes was stronger than pure drugs,and were positively correlated with drug concentration.6.The chick chorioallantoic membrane.The experimental results show that PAHDAA and PAHDAA-Cu revealed thatthe formation of new blood vessels were inhibitedincluding reduced vascular branches,thin and short forthe newly formed blood vessels.A slight hemolysis was observed.Conclusions1.PAHDAA and PAHDAA-Cu have a smaller cytotoxicity,and its anti-tumor mechanism may be through the inhibition of NF-kappa B nuclear transcription factor,and ultimately induce apoptosis.2.The PAHDAAand PAHDAA-Cu has a good ability to resist invasion and metastasis,and PAHDAA-Cu has a better effect.3.PAHDAA and PAHDAA-Cuhavethe ability to inhibit the formation of blood vessels,reduce the risk of metastasis of malignant tumors,andthat mayprovide a new direction for the development of new antiangiogenesis drugs.