| Objective: Malignant lymphoma is in the source of lymphaden and lymph tissue, it is a kind of malignant tumor about immune system. It is concerned with canceration about some kinds of immunocell which is happened of proliferation in lymphocyte in the process of immune response. Malignant lymphoma is one of the frequent malignant tumours in childhood. Combination chemotherapy is still the chief means about therapy in the nowadays. But the medicine which is frequently used about chemo often low the power of resistance and cause grave infect. This is one of important factor that effect the patients'long term survival. Therefore, it is very important to improve the patients'prognosis and quality of life about to find a kind of medicine which is high performance and low poison to cure lymphoma. Elemene is extracted in rhizome of curcuma aromatica salisb, which is an active component about anticancer. It is a new kind of anticancer drugs and is used to treated many kinds of tumor with its'distinct pharmacologic action. It has very powerful effect about to kill tumor cell and contain tumor to live. Elemene is wide about anticancer, high performance, security, and asepsis. It is no damage about the function of liver and kidney and is no bone marrow depression. It can elevate life quality about patients'. Telomerase is a special reverse transcriptase of nuclear ribonuclear protein, which is composed with RNA and protein. It includes human telomerase RNA (hTR), human telomerase reverse transcriptase (hTERT) and telomerase associated protein (TEP). The new expression of telomerase is very important about the process of escaped senescence about tumor cell and the activation of telomerase can level off the length of telomere and is even can make the cell gain immortalization. The masculine expression of telomerase activation is close correlation about genesis, development and the extent of cell differentiation about tumor. And it can be used for judging innocence, malignance and prognosis about tumor. Telomerase become one of foregone and the most broad-spectrum tumor markers. HTERT is a kind of reverse transcriptase, it has seven subunits. It is a catalytic subunit of telomerase activation. Further mechanisms to study and research telomerase activation is mostly regulate at transcriptional level. Therefore, hTERT is a determinative factor about the accommodation of telomerase activation. The activation about promoter of hTERT can detect in most kind of malignant tumors and it has upstair associativity with telomerase activation. Now, it is not yet seen about investigate and report on effect of elemene on proliferation, apoptosis, and human telomerase reverse transcriptase (hTERT) expression in human lymphoma cell line. In this study, in order to investigate the influence of elemene on proliferation, apoptosis, cell cycle distribution and hTERT expression in human malignant lymphoma cell line Raji. From this experiment we will approach about the effect of elemene on malignant lymphoma. And we can research possible anticancer mechanism of elemene though the influence about hTERT. Thus, theory evidence of elemene in clinical application will be provided.Methods: Human Burkitt's lymphoma cells Raji were cultivated in vitro and then treated with various concentration of elemene. Cell inhibitive rate was investigated by MTT assay. Cell cycle distribution and apoptosis rate in Raji cells were detected by Flow cytometry (FCM). Expression of hTERT mRNA in Raji cells was detected by RT-PCR.Results: 1 MTT showed: Cell inhibitive rate of Raji after treated with various concentrations of elemene for 24 hours were 5.24%,13.29%,25.11%,58.39% respectively. Cell inhibitive rate of Raji after treated with various concentrations of elemene for 48 hours were 23.43%,29.94%,37.23%,72.25% respectively. Cell inhibitive rate of Raji after treated with various concentrations of elemene for 72 hours were 28.80%,53.00%,68.97%,81.3% respectively. Absorbance light degree in experiment groups after treated Raji cells for 24h,48h,72h had statistical significance compared with control group (P<0.05), at the same time, there was statistical significance between concentration groups and time groups (P<0.05). 2 FCM showed: After Raji cells treated with various concentrations of elemene for 48 hours, the cell percentage in G0/G1 phase of test groups were(54.78±0.33)%,(59.89±0.27)%,(68.71±0.23)%,(74.78±0.26)% respectively; the cell percentage in S phase were(21.30±0.26)%,(19.73±0.28)%,(19.30±0.25)%,(18.18±0.20)% respectively; the cell percentage in G2/M phase were(23.89±0.18)%,(20.37±0.33)%,(12.04±0.21)%,(7.34±0.22)% respectively. The cell percentage in G0/G1, S, G2/M phase of control group were (50.30±0.48)%,(25.20±0.35)%,(24.51±0.30)% respectively. Compared with control group, cell cycle distribution in test groups of elemene existed a significant difference (P<0.05). By one-way ANOVA, cell cycle distribution had statistical significance among concentration groups (P<0.05). 3 FCM showed: Apoptosis peak emerged significantly after Raji cells treated with elemene for 48hours. Apoptosis percentage (AP) were (5.66±0.33)% , (6.24±0.36)%,(6.95±0.29)%,(17.20±0.27)% respectively; there was no or only low apoptosis peak in control group, AP in control was (2.20±0.34)%. There was remarkable difference about AP between experiment groups and control group. By one-way ANOVA, AP were significantly different among concentration groups (P<0.05). 4 RT-PCR showed: The correspondent expression of hTERT mRNA in Raji cells after treated with elemene for 48 hours at the concentrations of 10μg/ml, 20μg/ml,30μg/ml,40μg/ml were 0.82±0.022,0.72±0.020,0.50±0.016,0.47±0.019 respectively. The correspondent expression of hTERT mRNA in control group was 0.90±0.030. Experiment groups compared with control group about the correspondent expression of hTERT mRNA had statistical significance (P<0.05). The decreasing expression of hTERT mRNA existed statistical significance among concentration groups (P<0.05).Conclusions: 1 The survival of Raji cells was inhibited by elemene in vitro with a concentration and time dependent manners from 10-40μg/ml. 2 The number of Raji cells in G0/G1 phase was increased, while those in S and G2/M phase were decreased by the effect of elemene. Elemene induced cell cycle arrest in G0/G1 phase and inhibited the growth of Raji cells, which may be one of the mechanisms of antitumor. 3 In this study, before the cells were interfered with drugs, hTERT were highly expressed in Raji cells, which suggested that hTERT together with telomerase activation and infinite reproductive of tumor cells existed internal association. 4 The expression of hTERT in Raji cells can be downregulated by elemene in vitro with a concentration dependent manner, and apoptosis in Raji cells can be induced by elemene through the anomalism of telomerase activation from the downregulation of hTERT expression, which may be another possible anticancer mechanism of elemene.
|
PDF Full Text Request