Effect Of Zedoary Turmeric Oil And Some Components On Chronic Myelocytic Leukemia And Its Mechanism

Chronic myelocytic leukemia (CML), which make up twenty percent of all the leukemia, is very harmful for human health with wide age distribution and unfavourable prognosis. CML is characterized by the Philadelphia (Ph) chromosome creating a hybrid bcr/abl gene, which results from the reciprocal t(9;22)(q34;q11), with the ab1 proto-oncogene from 9q34 fused to the breakpoint cluster region (bcr) locus on 22q11. The expression product of bcr/abl, P210 can promote cell proliferation and inhibit cell differentiation and apoptosis through activating ras, Erk, jun kinase, PI3K and JAK-STAT with its tyrosine kinase. It is an important method to eliminate the bcr/abl rearrange gene for CML therapeuty and preventing drug resistant and recidivism. Herein, it is an significant topic to search novel targeted drug for the fusion gene eliminated.Zedoary Turmeric Oil (ZTO), which is extracted from the traditional Chinese medicine Zedoary, have antitumor effect with a little adverse effect. Curcumol and p-elemene are two principal component in ZTO. Although p-elemene can inhibit tumor growth and influence the expression of oncogene bc1-2, it trend to result in nausea, inappetence, fever, phlebitis and severe pain, etc., which restrict its clinical application markedly. Curcumol is the maximum component in the ZTO, which was once reported to cure uterine cervix cancer with good curative effect. So, it is significant to research the antitumor effect of curcumol and its mechanism.Curcumol was extracted by crystallization and recirculation method. Liposome of curcumol or ZTO was prepared and identified with spectrophotometry. Human CML cell line K562 and mouse lymphocytic leukemia cell line L1210 were used to study the effect of curcumol, P-elemene and ZTO and their mechanism in this experiment. Cell proliferation and growth curve were observed with MTT assay. Cell apoptosis, differentiation were determinatd with Wright-Gimsa staining, laser scanning confocalmicrdscopy, NBT-reduction test and cytochemical staining techniques. Bcl-2 mRNA, bax mRNA and bcr/abl mRNA expression levels were measured by RT-PCR method. Micronucleus test was used to observe the chromosome damage.Results:1. Curcumol was extracted from ZTO and the yield rate was 2.574%. The diameter ranges of curcumol liposome and ZTO liposome were between 100 and 1000 nm. Envelopment ratios of curcumol liposome and ZTO liposome were 86.2% and 74.5%, respectively.2.Curcumol and ZTO could suppress the proliferation of K562 and L1210 cell significantly with dose- and time-dependence.3.Curcumol 10g/ml induced K562 cell chromatin aggregates and karyorrhexis. The apoptosis rates in curcumol 30 g/ml group were 28.7%, 50.0%, 78.3% at 24h, 48h and 72h, respectively.3.Curcumol (10, 30g/ml) induced K562 cell to differentiate to mature stage.4. Curcumol (30, 100 g/ml) decreased bcr/abl mRNA expression and increased bax mRNA expression significantly (p<0.05). Bcl-2 mRNA was not detected in all groups.5.Micronucleus test indicated that curcumol (30, 100 g/ml) decresed micronucleus rates markedly after 48h compared with control group. ZTO had no significant effect.Conclusion:1 . Curcumol was successfully extracted and the liposome was prepared qualifiedly.2. Curcumol and ZTO could suppress the proliferation of K562 and L1210 cell significantly with dose- and time-dependence.3. Curcumol could induce K562 cell apoptosis, which related to bc1-2 and bax pathway.4, Curcumol could repair bcr/abl fusion gene, induce tumor cell apoptosis and promote differentiation as well, which might be the mechanism of inhibiting leukemia cell proliferation.