Dissertation
Dissertation > Medicine, health > Chinese Medicine > Of Pharmacy > Pharmacology > Chinese medicine Experimental Pharmacology

Effects of the Effective Component of Panax Ginseng on the Proliferation of Leukemia KG1α Cells

Author LiuXiaoYan
Tutor ChenDiLong;LiJing
School Chongqing Medical University
Course Human Anatomy and Embryology
Keywords Active ingredients of ginseng KG1α cells Inhibition of proliferation Apoptosis
CLC R285.5
Type Master's thesis
Year 2011
Downloads 31
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The chemotherapy cancer is one of the fastest growing areas in clinical therapeutics, chemical anticancer drugs developed so far are mostly cytotoxic drugs, they can not be side effects and poor selectivity obstacles grams taking poison. On the one hand, many tumor cells also varying degrees of anti-tumor chemical drugs are not sensitive to low doses is difficult to suppress the malignant proliferation of tumor cells, high dose will cause serious injury to body tissues and organs, on the other hand the hematopoietic and immune system of the body so they even lower doses can also lead to the irreversible destruction of these systems are extremely sensitive to chemotherapeutic drugs. One of the core content of the modernization of Chinese medicine is a theoretical system of Traditional Chinese Medicine to provide the theoretical basis of modern science and experimental evidence. Is worth noting that the Chinese medicine in the treatment of cancer, has accumulated a wealth of information, the good effect of traditional Chinese medicine is increasingly recognized by people. Initial screening of anti-tumor medicine through its structural analysis in modern biomedical technology, analog modification, pharmacological and toxicological in-depth research, entirely possible to develop a new, highly efficient, low toxicity of anticancer drugs. Objective: To study ginseng saponins (TSPG) and Ginsenoside Rh2 (S) on human acute myeloid leukemia cell lines KG1α proliferation and cell cycle and apoptosis, and using immunocytochemistry and molecular biology The method validation TSPG interfere with KG1α growth possible mechanisms. Methods: logarithmic phase KG1α cells, adjust the density of 5 × 10 8 / L, blank control group were given conventional culture; the ginsenosides the TSPG group were added 0.1,0.2,0.4,0.6, 0.8 g / L TSPG. MTT colorimetric determination of cell viability, Wright's staining and light microscopy, electron microscopy morphological changes. Flow cytometry determination of changes in the distribution of cell cycle and cell apoptosis rate. Immunocytochemical methods to detect changes in the expression of β-catenin and of NF-κBp65 protein in the cells. Β-catenin in the semi-quantitative RT-PCR detection cells, NF-κBp65 CyclinD1 and of TCF4 the mRNA level changes. Semi-quantitative Western blot detection cell β-catenin, NF-κBp65, CyclinD1 and TCF4 protein levels change. Take the the logarithmic phase KG1α cells adjust the density of 5 × 10 8 / L, the control group were given conventional culture; the ginsenosides monomer the Rh2 (s) group were added 0.03,0.035,0.04, 0.045,0.05 g / L Rh2 (s). MTT colorimetric determination of cell viability, Wright's staining and morphological changes of light electron microscopy. Flow cytometry determination of changes in the distribution of cell cycle and cell apoptosis rate. Results: Ginseng total saponins the TSPG in vitro proliferation KG1α, has a significant inhibitory effect. 0.1-0.8g / L TSPG concentration range, the inhibition was concentration dependent, to reach half of the inhibition rate of the cells in the concentration of 0.4g / L at, and 48h inhibition rate reached its peak. Compared with the blank control group cells into a group gathered growth becomes decentralized growth, significantly reduced the number of cells; cells were differentiated to produce mitotic; Some of apoptosis, cytoplasmic produced 0.4g / L TSPG after 48h, visible large number of vacuoles and apoptotic bodies; KG1α cell cycle distribution was no significant change; significantly higher rate of apoptosis; Immunocytochemistry showed reduced expression of intracellular β-catenin and of NF-κBp65 protein, the expression region from the nucleus to the extracellular film transfer; RT-PCR semi-quantitative detection of cells in β-catenin, the NF-κBp65, of CyclinD1 and TCF4 mRNA levels reduce. Semi-quantitative Western blot detection cells in β-catenin, NF-κBp65 TCF4 protein levels decreased. Ginsenoside Rh2 (s) the in vitro proliferation KG1α has a significant inhibitory effect. In the concentration range of 0.03-0.05g / L Rh2 (s), the inhibition was concentration dependent, to reach half of the inhibition rate of the cells in the concentration of 0.04g / L at, and peaked at 48h inhibition rate. Compared with the blank control group, 0.04 g / L the Rh2 (s) role 48h, can also be found cells was dispersed growth, significantly reduced the number; Some cell differentiation to produce mitotic, apoptosis, cytoplasmic vacuoles and apoptotic bodies; blank control group and were cultured in vitro after 48h 0.04 g / L Rh2 (S) group G 0 / G 1 phase and G 2 < / sub> M phase cell distribution results compared, the difference was statistically significant. S phase difference of the two groups was not statistically significant. Rh2 (S), the rate of apoptosis after treatment was significantly higher. Conclusion: TSPG TSPG can inhibit KG1α cell proliferation, induction of apoptosis, and the mechanism may be related to reduced β-catenin TSPG suppression KG1α cell proliferation the NF-κBp65, of CyclinD1 and of TCF4 the mRNA level, inhibition of β-catenin, The NF-κBp65 TCF4 protein. Ginsenoside Rh2 (S) can inhibit the KG1α cell proliferation KG1α arrest in G0/G1 phase. FCM measured rate of apoptosis, Rh2 (S) can induce apoptosis. TSPG experimental study, the rate of apoptosis in the control group (8.49 ± 0.66)% of TSPG group apoptosis rate (23.56 ± 0.99)%; Rh2 (S) in the experimental study, the control group, the apoptosis rate ( 2.58 ± 0.40)% apoptosis rate of Rh2 (S) group (10.44 ± 3.17)%. TSPG cause apoptosis rate of change of amplitude Rh2 (S) caused the rate of change of the rate of apoptosis was not statistically significant. Shows, Rh2 (S) the ability to induce apoptosis and TSPG quite, but its mechanism of action with TSPG may need further study.

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