Study The Effect Of Matrine On Proliferation And Mechanism Of It's Anti-cancer In Extrahepatic Cholangiocarcinoma Cell QBC-939

Objective: To investigate the effect of matrine on the proliferation and apoptosis of human extrahepatic cholangiocarcinoma cell QBC-939 and try to expound the mechanism of it's anti-cancer.Methods: Human extrahepatic cholangiocarcinoma cell QBC-939 was co-cultured with matrine in different concentrations. MTT essay was used to measure the level of the proliferation of cell QBC-939 and the effect of matrine on cell cycle and cell apoptosis of cell QBC-939 was observed by FCM. The cellular content of p27kip1 was measured with immunohistochemical method and image analysis. The morphological changes of cell were found by light microscopy and electronic microscopy.Results: The proliferation of cell QBC-939 was obviously inhibited by matrine in a dose and time dependent manner. After treated by 0.25~1.5g/L matrine for 48h, the cell inhibition rate increased gradually, respectively 21.23%,37.13%,57.32%,66.34%ï¼›and the comparing of OD value between control group and experimental group was significant(p<0.01).The matrine concentration had a negative correlation with OD value(r=-0.957).After incubation with 1.0g/L matrine for 24~96h,the comparing of OD value between control group and experimental group in different time interval was significant (p<0.01), and the cell inhibition rate increased gradually, respectively 37.21%,57.32%,67.45%,72.32%. The outcome of FCM showed that there was no apoptosis in cell QBC-939 induced by matrine, but matrine can effect the location of cell cycle. After treated by 0.25~1.5g/L matrine for 48h ,the percentage of cell number in G0/G1 phase was 56.15%,60.71%,64.07%,70.89%, and in S phase was 29.70%,26.73%,25.35%,23.65%,21.39ï¼…,and in G2/M phase was 14.15%,12.56%,10.58%,8.73%,7.72%.With the increasing of concentration, matrine has a positive correlation with cell in G0/G1 phase (r=0.969),but negative correlation with cell in S and G2/M phase (r=-0.901,r=-0.922). Matrine could make cell cycle arrested in G0/G1 phase. The morphological changes of apoptosis were also not found by electronic microscopy, but cell was filled with round and transparent granules. After incubation with 0.25~1.5g/L matrine for 48h, the cell number decreased gradually by light microscopy. The content of p27kip1 was depended on the dosage of matrine. The matrine concentration had a negative correlation with p27kip1 grey level ( r=-0.973).Conclusion: Matrine can inhibit the proliferation of human extrahepatic cholangiocarcinoma cell QBC-939. The mechanism of it's anti-cancer was cell cycle arrest ,not induced the apoptosis of the cell. The up-regulation of p27 might be correlated with cell cycle arrest.