Experimental Studies On Different Concentrations Of Bupleuri Combine Curcuma Decoction In The Inhibition Of HepG₂Cell Lines

[Objective]Different concentrations of Bupleuri combine Curcuma Decoction were putted into invitro HepG2cell lines, and then observe the suppression effect of HepG2proliferation and expression of Ki67. Then compare the suppressor effect with Bupleuri Decoction and Curcuma Decoction respectively, and from the ancient Chinese medicine classics to explore the prospects for its tumor suppressor mechanism and prospects in the clinical treatment of liver cancer.[Methods]Hepatocarcinoma HepG2cells were divided into5groups:the control group, the groups treated with5-Fu and Bupleuri combine Curcuma Decoction35mg/ml,·28mg/ml,21mg/ml,14mg/ml,7mg/ml. Tested the suppression ratio of cells by using MTT, tested the growth of cells by using growth curve method, and test the expression of Ki67in cells by using immunohistochemistry Compare the suppression ratio of cells between the Bupleuri combine Curcuma Decoction (21mg/ml,14mg/ml) and Bupleuri Decoction (10.5mg/ml,7mg/ml), Curcuma Decoction (10.5mg/ml,7mg/ml) by using MTT.[Results]Bupleuri combine Curcuma Decoction can significantly inhibit the growth of HepG2and make an induction of apoptosis on the HepG2.The MTT test showed that Bupleuri combine Curcuma Decoction could make human liver cancer cell proliferate in a dose--effect relationship--the more concentration, the more incidence of inhibition. But the suppression ratio of35mg/ml concentration was lower than28mg/ml concentration. The suppression ratio of5-Fu and35mg/ml,28mg/ml,21mg/ml,14mg/ml,7mg/ml Bupleuri combine Curcuma Decoction were75.60%,95.27%,97.01%,81.66%,55.08%,7.65% respectively. Its50%inhibitory concentration(IC50)is about]3.03mg/ml,90%inhibilory concentration(IC90)is about22.27mg/ml. The growth curve showed that the cells in all groups except the control group were suppressed, and the suppressed action in the groups treated with35mg/ml,28mg/ml,2]mg/ml,14mg/ml Bupleuri combine Curcuma Decoction was better than the group treated with7mg/ml. The Ki67index of control group, the groups treated with35mg/ml,28mg/ml,21mg/ml,14mg/ml,7mg/ml Bupleuri combine Curcuma Decoction were94.73±2.79,71.67±3.56,70.53±3.31,76.03±3.02,79.53±3.55,83.3±2.61respectively, there was a significant statistical differences between the groups treated with Bupleuri combine Curcuma Decoction(P<0.01). By the concentration gradient pairwise comparisons Bupleuri combine Curcuma decoction of35mg/ml,28mg/ml group showed no statistically significant (P>0.05), other groups were significantly different (P <0.01). The suppression ratio of21mg/ml,14mg/ml, Bupleuri combine were81.66%,55.08%, significant higher than the10.5mg/ml,7mg/ml Bupleuri Decoction (P<0.01). The Curcuma Decoction has no significant killing effect on the HepG2cell lines, but showed some growth-promoting effect, the suppression ratio of10.5mg/ml,7mg/ml Curcuma Decoction is-7.65%and-8.26%, but there is no statistical significance to the control group (P>0.05).[Conclusions]Bupleuri combine Curcuma Decoction could suppress the growth of the HepG2cell lines, kill the cells and depress the expression of Ki67in cells. It was a concentration-effect relationship. The Curcuma Decoction has no significant killing effect on the HepG2cells, but combine with bupleuri decoct together, the tumor suppressor effect was significantly better than Bupleuri Decoction only. The Bupleuri combine Curcuma Decoction may have a potential sensitizer, synergies effect on liver cancer treatment.