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Inhibitory Effects of Ligustrazine and Matrine on Infiltration and Metastasis of Leukemia Cells

Author: ZhangWei
Tutor: DaiBiTao
School: Chongqing Medical University
Course: Pediatrics
Keywords: Ligustrazine Matrine Leukemia Infiltration Metastasis
CLC: R285.5
Type: Master's thesis
Year: 2008
Downloads: 127
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Abstract


Background:Leukemia, which threats the patients’lives severely and affects the patients’quality of living, is the malignant tumor from hematopoietic system and is the most common malignant tumor in children. With the knowledge of the genesis and development of leukemia and the improvement of combined chemotherapy, the complete remission rate of leukemia has been heightened. Leukemia cells can infiltrate into central nervous system or testis. If this happens, it will prolong the time of therapy and lead to the recurrence for complete remission patients. In clinic, the main way to prevent and treat it is to increase the dose of chemotherapeutics. Unfortunately, increasing the dose of chemotherapyeu- tics also has the side effects of chemotherapeutics and it will increase the cost on the therapy for the patients. So, it is important to look for available chemotherapeutics which can inhibit the infiltration and metastasis of leukemia cells effectively. Recently, traditional Chinese drugs have been paid close attention to because of their special pharmacological action. It has been reported widely that Ligustrazine and Matrine can inhibit cell proliferation, induce cell differentiation, facilitate apoptosis, and reverse multidrug resistance of tumors. Our previous study has confirmed that Ligustrazine can inhibit the infiltration and metastasis of HL-60 cells effectively; but other reports on the inhibitory effects of Ligustrazine and Matrine on infiltration and metastasis of leukemia cells haven’t been seen.Objective:To study the inhibitory effects of Ligustrazine and Matrine in low, medium and high concentration on adhesion, migration, invasion and expression of matrix metalloproteinases of HL-60 cells and Jurkat cells; and to investigate their possible mechanism of action.Methods:HL-60 cells and Jurkat cells were cultured in vitro. The MTT assay was used to choose the best concentration of Ligustrazine and Matrine. Jurkat cells were treated by 0.05g/L, 0.1g/L and 0.15g/L Ligustrazine, and HL-60 cells and Jurkat cells were treat by 0.1g/L, 0.15g/L and 0.2g/L Matrine. Then cell adhesion assay, cell migration assay and cell invasion assay were used to observe the inhibitory effects of Ligustrazine and Matrine on adhesion, migration and invasion of leukemia cells; and RT-PCR were performed to evaluate the MMP-2 and/or MMP-9 mRNA expression levels.Results:1. Treated by Ligustrazine in low, medium and high concentration for 48 hours, cell survival rates of Jurkat were (90.1±1.1)%, (82.8±1.8)% and (74.3±2.2)% respectively. Treated by Matrine in low, medium and high concentration for 48 hours, cell survival rates of HL-60 were (88.2±0.9)%, (81.6±1.4)% and (74.9±2.1)% respectively; and Jurkat were (87.4±2.1)%, (81.0±1.2)% and (73.4±1.2)% respectively;2. Treated by Ligustrazine in low, medium and high concentration, cell adhesion rates of Jurkat for 1 hour were (86.8±0.4)%, (79.2±1.9)% and (71.7±2.2)% respectively; cell migration rates of Jurkat for 8 hours were (87.5±0.5)%, (75.1±0.8)% and (61.9±1.2)% respectively; and cell invasion rates of Jurkat for 48 hours were (81.4±2.2)%, (66.0±3.6)% and (50.0±5.3)% respectively;3. Treated by Matrine in low, medium and high concentration, cell adhesion rates of HL-60 for 1 hour were (85.8±0.5)%, (73.5±2.0)% and (61.9±1.3)% respectively; cell migration rates of HL-60 for 8 hours were (82.9±1.1)%, (70.9±0.6)% and (60.1±2.5)% respectively; and cell invasion rates of HL-60 for 48 hours were (80.2±2.3)%, (63.9±3.2)% and (48.8±2.2) % respectively;4. Treated by Matrine in low, medium and high concentration, cell adhesion rates of Jurkat for 1 hour were (88.1±0.6)%, (79.9±1.2)% and (70.0±1.6)% respectively; cell migration rates of Jurkat for 8 hours were (88.3±1.0)%, (74.7±2.2)% and (61.9±2.2)% respectively; and cell invasion rates of Jurkat for 48 hours were (84.8±1.7)%, (70.4±2.2)% and (53.4±4.1)% respectively;5. Treated by Ligustrazine in low, medium and high concentration for 48 hours, the expression rates of MMP-9 mRNA in Jurkat were (73.2±4.0)%, (51.3±2.3)% and (46.7±1.6)% respectively. Treated by Matrine in low, medium and high concentration for 48 hours, the expression rates of MMP-2 mRNA in HL-60 were (88.2±7.4)%, (87.8±9.2)% and (85.7±8.5)% respectively; those of MMP-9 mRNA in HL-60 were (78.3±3.4)%, (67.8±6.4)% and (66.3±4.4)% respectively; and those of MMP-9 mRNA in Jurkat were (82.4±6.5)%, (72.2±8.7)% and (66.1±6.5)% respectively;6. The expression of MMP-9 was obvious but MMP-2 was not in Jurkat cells; and the expressions of MMP-2 and MMP-9 were obvious in HL-60 cells;7. There was positive correlation between the capability of invasion and the expression of MMP-9 in Jurkat cells; and there was positive correlation between the capability of invasion and the expression of MMP-2 and MMP-9 in HL-60 cells.Conclusion:1. Ligustrazine and Matrine can inhibit the proliferation of HL-60 cells and/or Jurkat cells, and the inhibitory effects are dose and time dependent. In this study, the best concentrations of Ligustrazine are 0.05g/L, 0.1g/L and 0.15g/L, and those of Matrine are 0.1g/L, 0.15g/L and 0.2g/L;2. Ligustrazine in concentration of 0.05g/L, 0.1g/L and 0.15g/L can inhibit the adhesion, migration and invasion of Jurkat cells; and the inhibitory effects are dose dependent;3. Matrine in concentration of 0.1g/L, 0.15g/L and 0.2g/L can inhibit the adhesion, migration and invasion of HL-60 cells; and the inhibitory effects are dose dependent;4. Matrine in concentration of 0.1g/L, 0.15g/L and 0.2g/L can inhibit the adhesion, migration and invasion of Jurkat cells; and the inhibitory effects are dose dependent;5. Ligustrazine in concentration of 0.05g/L, 0.1g/L and 0.15g/L can down- regulate the expression of MMP-9 mRNA in Jurkat cells. Matrine in concentration of 0.1g/L, 0.15g/L and 0.2g/L can down-regulate the expression of MMP-2 and MMP-9 mRNA in HL-60 cells; and down-regulate the expression of MMP-9 mRNA in Jurkat cells;6. There is obvious expression of MMP-9 mRNA but not for MMP-2 in Jurkat cells; and there are obvious expressions of MMP-2 and MMP-9 mRNA in HL-60 cells;7. By down-regulating the expressions of MMP-2 and/or MMP-9 mRNA, Ligustrazine and Matrine can inhibit the infiltration and metastasis of leukemia cells.

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