Influence of traditional medicine prescription on Flt3 and N-ras mRNA expression in leukemic stem cells
10.3969/j.issn.1673-8225.2009.40.033
- VernacularTitle:中药复方对急性髓细胞性白血病干细胞Flt3和N-ras基因表达的影响
- Author:
Ruirong XU
;
Xiaoling WANG
;
Jingyi WANG
;
Xing CUI
;
Yan WANG
;
Lizhen LI
- Publication Type:Journal Article
- From:
Chinese Journal of Tissue Engineering Research
2009;13(40):7969-7973
- CountryChina
- Language:Chinese
-
Abstract:
BACKGROUND:There is a group of leukemic stem cell (LSC) in patients with leukemia. These malignant stern cells, although rare, but self-renewal, with a certain degree of differentiation is the existence of leukemic cells and causes of their continued proliferation. Intervention of leukemia-specific targeted treatment drugs has become a research hotspot in recent years. OBJECTIVE: To investigate the influence of traditional medicine compound on expression of flt3 and N-ras mRNA in LSCs. DESIGN, TIME AND SETTING: A randomized, grouping experiment was performed at the Affiliated Hospital of Shandong University of Traditional Chinese Medicine from September 2007 to December 2008.PARTICIPANTS: A total of 50 cases of acute myelocytic leukemia admitted to Department of Hematology, Affiliated Hospital of Shandong University of Traditional Chinese Medicine and Qilu Hospital of Shandong University between 2006 and 2007 were selected, including 5 cases of type FAB, 15 of M2, 9 of M4 and 21 of M5.METHODS: Original prescription: Huangqi, Hedyotic diffusa, Xiaoji, Taizishen, Banzhilian, Pugongying, Rehmannia dride rhizome, Huangjing, Nuzhenzi, ecliptae herba, Tiandong, Maidong, Baizhu, Fuling, Liquorice root. Fuzheng prescription: Huangqi, Hedyotic diffusa, Xiaoji, Banzhilian, Pugongying, ecliptae herba. Preparation: the above-mentioned three groups of drugs were prepared into 1 g/mL medicine liquid by Laboratory of Chinese Drug Preparation of Affiliated Hospital of Shandong University of Traditional Chinese Medicine. When it was negative in sterility test, the liquid was filtrated and sterilized. 5 mL bone marrow was separately harvested from M1, M2, M4, and M5 leukemia patients, diluted and placed in lymphocyte isolation solution. LSCs were purified by magnetic activated cell sorting and flow cytometry. The cell concentration was adjusted to 2×108/L and divided into 4 groups: control (no treatment), and three experimental groups (treated separately with original, Fuzheng, Quxie prescriptions at final concentration of 100 mg/L). The cells were cultured for 48 additional hours. MAIN OUTCOME MEASURES: RT-PCR was used to detect the mRNA expressions of Flt3 and N-ras in LSCs. RESULTS: The expression of flt3 in three experimental groups was significantly decreased compared with control group (P < 0.05), in particular, original prescription group decreased the most (P<0.05), and no significant difference was found between Fuzheng and the Quxie prescription groups (P>0.05). The expression of N-ras in four groups was similar to Flt3. CONCLUSION: Flt3 and N-ras mRNA were overexpressed in LSCs. Chinese medicine original prescription and its Fuzheng or Quxie prescriptions decreased Flt3 and N-ras mRNA expression.