| As one of the important anti-tumor therapies, chemotherapy had the advantage of effective and quick response, and the disadvantage of serious adverse reactions for it had a harmful impact on healthy cells. So it is necessary to innovate in new anti-tumor drugs. As a Traditional Chinese Medicine, Zedoary Turmeric Oil(ZTO) is the volatile oil from Curcuma wenyujin, and composed of curcumol, β-elemene and so on. Among them, the content of curcumol is the highest. Curcumol was reported to be effective in anti-uterine cervix cancer with few adverse reactions. Although β-elemene had anti-tumor effects as well, it tended to bring many adverse reactions which hindered its wide use. Curcumol extracted from ZTO, was used to study the anti-hepatoma and anti-colon carcinoma effect and its mechanism which is expected to provide theoretical support for the clinic use of curcumol. METHODS:Curcumol was extracted by method of crystallization and recirculation and was identified by spectrophotometry, then curcumol and ZTO were prepared to liposome. Proliferation of HepG2 cell and HT-29 cell in vitro were observed with curcumol, ZTO and β-elemene by the means of growth curve modeling and MTT assay. Apoptosis of cells was observed by staining with Hoechst 33258/PI and laser confocal scanning microscopy(LCSM). Expression of COX-2 mRNA and VEGF mRNA of HepG2 cell and HT-29 cell were investigated by RT-PCR, and expression of COX-2 and VEGF were observed by immunohistochemistry technique in mice.RESULTS:1. Curcumol was extracted from ZTO and the yield of curcumol was 2.574%. The diameter range of curcumol liposome and ZTO liposome was 100 to 1000nm. Envelopment ratios of curcumol liposome and ZTO liposome were 86.2% and 74.5%, repectively.2. Curcumol 27μg/ml decreased the quantity of HepG2 cell at 7th day and the quantity of HT-29 cell at 6 th, 7 th day significantly. Curcumol 9~80μg/ml inhibited proliferation of HepG2 cell and HT-29 cell with by MTT assay with dose- and time- dependence, and ZTO and β-elemene decreased proliferation of HepG2 cell and HT-29 cell. Growth of tumor in H22 mice was inhibited by curcumol 100mg/kg. And inhibition ratio was 51.3%(p<0.05), which was not different from inhibition ratios of ZTO and β-elemene significantly(p>0.05).3. Curcumol 27μg/ml, ZTO 27μg/ml and β-elemene10μg/ml induced HepG2 cell and HT-29 cell apoptosis after 48h. LCSM results showed that apoptosis ratio of HepG2 cell and HT-29 cell was higher by the treatment of curcumol, ZTO and β-elemene significantly (p<0.05). The apoptosis rates in HepG2 cell were 30.2%, 12% and 25.2%; The apoptosis rates in HT-29 cell were 38%, 22.4% and 44%.4. VEGF and COX-2 were expressed in the cytoplasm in tumor tissue. Expression of COX-2 and VEGF were diminished by curcumol significantly(p<0.01), and inhibition ratio of curcumol was better than ZTO, reached to 43.% (p<0.01) and 31.5%(p<0.01), respectively.5. Expression of VEGF mRNA of HepG2 cell and HT-29 cell were discreased by curcumol 9, 27, 80μg/ml, ZTO 9, 27, 80μg/ml and β-elemene 10, 30μg/ml significantly (p<0.05).6. Expression of COX-2 mRNA of HepG2 cell was reduced by curcumol 80μg/ml, ZTO 80μg/ml and β-elemene 10, 30μg/ml significantly(p<0.05). Expression of COX-2 mRNA of HT-29 cell was diminished curcumol 27,80μg/ml and β-elemene 10, 30μg/ml significantly(p<0.05). ZTO had no abvious effects on that(p>0.05).CONCLUSION:1. Curcumol was extracted from ZTO successfully, and Curcumol and ZTO were prepared to liposome.2. Proliferation of HepG2 cell and HT-29 cell in vitro were inhibited with curcumol with dose- and time- dependence significantly.3. Proliferation of tumor in H22 mice was inhibited significantly.4. Apoptosis of HepG2 cell and HT-29 cell in vitro were inducted by curcumol.5. Expression of VEGF and COX-2 in tumor tissue of H22 mice in vitro were inhibited by curcumol. 6. Expression of VEGF mRNA and COX-2 mRNA of HepG2 cell and HT-29 cell n vitro were inhibited by curcumol. |
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