The Influence Mechanism Of Breviscapine To The Ability Of HePG2 Cell Biological Character Research

Objective:To study the influence of breviscapine in primary liver cancer cell proliferation, migration, invasion and influence in the process of apoptosis, and Jak2/STAT3 signaling pathway plays role in the process of mechanism research.Methods:(1) respectively in HepG2 liver cancer cells and normal BRL3a liver cells by 0 ug/ml,1 ug/ml,3 ug/ml,9 ug/ml,27 ug/ml,81 ug/ml concentration gradient to join breviscapine, to 1 x 106/a vaccination in 6 orifice and density close to 80%, the inspection and analysis was determined by MTT method to the breviscapine influence on HepG2 cells and BRL3a liver cell inhibitory concentration 50 (IC50) and proliferation after cultured 24 hours;(2) the experiment is divided into HepG2+Bre and HepG2 group, then respectively join the same amount of media and culture for 24 h, detecting the number of HepG2 cell migration of Transwell little indoor;(3) the experiment is divided into HepG2, HepG2+Bre group, used liquid move cross straight horizontal lines, cultivated 24 h and measured respectively 0h,12 h,24 h cell scratch distance;(4) the logarithmic phase of HepG2 cells divided into HepG2, HepG2 +Bre group,and respectively inoculated to 6 orifice to cultivate 24 h, detect the number of each group apoptosis cells with Tunel kit;(5) using Western blot method to detect the protein expression of Bax, the Bcl-2, Caspases3, Jak2 and Stat3 and p-Stat3 respectively in HepG2+Bre, HepG2 cell group, BRL3a+Bre and BRL3a liver cell group.Results:(1) determined by MTT experiment results show that by the various concentration of breviscapine processing for 24h,the breviscapine inhibitory concentration of HepG2 cell is13.68ug/ml, and it can inhibit HepG2 cell proliferation by different degree of (P<0.05), and the inhibition rate assumes the concentration dependence;While for BRL3a liver cell, the inhibitory concentration cannot be tasted and it’s proliferation is not significantly inhibited (P>0.05);(2) the Transwell experimental results show that the invasion HepG2 cell number in HepG2+Bre group small indoor was obviously less than HepG2 group (P<0.05);(3) the scratches experimental results showed that the scratches from breviscapine processing of HepG2 cells in 12 h,24 h had no significant change (P>0.05), the scratches from HepG2 cells without breviscapine processing significantly reduced (P<0.05);(4) Tunel experimental results show that the number of breviscapine processing of HepG2 group was dyed brown significantly greater than the number of cells without breviscapine of HepG2 group (P<0.05);(5) the analysis and testing results of Western blot experimental show that:the expression of Bcl-2, Jak2 and Stat3 and p-Stat3 protein in HepG2 group was higher than BRL3a liver cell group (P<0.05), while the expression of proteins Bax, Caspase3 significantly below than BRL3a liver cells (P<0.05); in HepG2 cells processing with breviscapine, the expression of Bax and Caspases3 protein increased significantly (P<0.001), while the the expression of Bcl-2, Jak2 and Stat3, p-Stat3 protein decreased significantly (P<0.001,P<0.01);in BRL3a liver cells treated by breviscapine,the the expression of Bax, Caspases3, Bcl-2,Jak2 and Stat3 and p-Stat3 protein compared with without breviscapine processing BRL3a liver cells has no significant change (P>0.05).Conclusions:(1)breviscapine promote HepG2 liver cancer cell apoptosis by inhibiting the Bcl-2, promote Bax, Caspase3 expression; (2)breviscapine can inhibit the activation of Jak2,Stat3 and p-Stat3 proteins of HepG2 cells thus to inhibit the proliferation of HepG2 cells, reduce its ability of migration and invasion; (3)breviscapine may inhibit JAK2/STAT3 signaling pathway to promote HepG2 apoptosis, inhibit the proliferation, migration and invasion of HepG2 cell.