High Through Put Screening Of Anti-liver Cancer Extrations Of Chinese Traditional Drugs And Study On Their Action Mechanisms

In the experiment, 1000 kinds of Chinese traditional drugs for curing liver cancer were studied. Through extracting of ethanol(ethanol: acid ester=6:4) and water(methyl alcohol: water=8:2).The FlashMaster II flash chromatography system is adopt automated purification method. By this apparatus, a library of 20000 chinese triditional drug extraction was established, using the drug screening MTT method we had screened the library to found 3 drugs'IC50 under 10μg/mL, then purified them to get bio-activite pure compounds.LCID-1 get from the Chinese traditional drugs high throughput screening by method of MMT .we studied its action on HepG2 cells and probe into action mechanism, offered the data for next drug exploiture.Cultured human liver cancer cell line HepG2 was treated with LCID-1. The changes of cell growth, proliferation and cell cycle were measured by MTT and FCM. Using microscope observed the variety of cell conformation; Effect of LCID-1 on apoptosis was detected by TUNEL. Transcription of Bcl-2 and Ki67 mRNA was measured by RT-PCR; Expression of bcl-2 and Ki67 was detected by immunogistochemistry. The main results were as follows:As a result:①The result of MTT showed, following treatment with LCID-1 under concentration of 0.01,0.1,1,10,100,1000μg/mL, the growth inhibition rate of liver cancer cell line was 6.15%,30.58%,52.72%,70.64%,96.26%,98.05%, respectively.Under the same conditions , the growth inhibition rate of the small rat normal liver cell were 5.92%,23.29%,41.94%,58.71%,81.59%,95.20% respectively.It shows that LCID-1 has less toxicity to human normal cells than to liver cancer cells.The IC50 of LCID-1 treating HepG2 cell is 5.5μg/mL,2.5μg/mL,0.9μg/mL at 24h,48h,72h.②FCM indicated that S phase distribution increased with the increase of drug concentration.③TUNEL result indicated 32.7﹪±4.8﹪tumor cells had the characteristics of apoptosis after LCID-1 treatment, while no apoptosis was found in HepG2 cells before treatment.④RT-PCR indicated Bcl-2 and Ki67 mRNA over expressed in HepG2 cells in control, and the expression was downregulated after LCID-1 treatment (P<0.05).⑤Immunohistochmistry staining showed overexpression of bcl-2 and Ki-67 in HepG2 cells was downregulated by LCID-1.The experimentation in vitro indicated:LCID-1,the Chinese traditional drug extraction has great merit of candidate drug to cure liver cancer.