Anti-Proliferative Effect And Mechanism Of Total Flavonoids Extracted From Juglans Mandshurica Maxim. On Hepg2 Cells Based On Microfluidic Chip

Juglans mandshurica Maxim, a traditional Manchu herbal medicine in China, has been used to treat or prevent various diseases. Total flavonoids, the primary chemical constituent of J. mandshurica (HH), possesses several bioactivities including anti-tumour. This study aimed at investigating the anti-liver cancer effect of HH in vitro through microfluidic chip, and expounding the molecular mechanism based on AKT relevant pathway. A multifunctional integrated microfluidic chip device, which can simulate the real microenvironment in the human body, was engineered to estimate the anti-liver cancer effect and mechanism of HH. The HepG2 cells cultivated in chip were treated with different concentrations of HH, and then proliferation inhibition was analyzed. Flow cytometry was used to assess cell apoptosis and cell cycle. Q-PCR and ELISA were performed to detect the expressions of genes and proteins on AKT-mTOR signaling pathway. The results demonstrated that HH inhibited the growth of HepG2 cells in dose-dependent and time-dependent manner, induced apoptosis and blocked cells cycle (G0/G1 phase) of HepG2 cells in a dose-dependent way. Gene and protein analysis revealed that the expression of PTEN mRNA and CASP-9, BAD proteins were significantly up-regulated while VEGF, AKT, mTOR mRNAs and VEGF, HIF-1 alpha proteins were down-regulated by HH treatment. These findings based on microfluidic chip provided the first experimental evidence confirming the efficacy of HH in anti-liver cancer via AKT-mTOR signaling pathway and paving the way for the broader use of HH in clinic.