Effect And Mechanism Of The Qi-tonifying And Astringing Method Interfering The Malignant Tumor Proliferation And Metastasis

Objective:Qi-tonifying and Astringing drugs are the drugs which are generally used to treat malignant tumors. The therapeutical effect of the drugs is related to the inhibition of tumor proliferation and metastasis, and anti-angiogenesis in tumor. Qi-tonifying and Astringing drugs can tonifying qi and inducing astringency to cancerous toxin, they also can extending the lives and improving the survival qualities of patients with cancer. This issue selects Astragalus and Agrimony which are usually used in clinical as the objects of study. Studies would be done on the influences of Qi-tonifying and Astringing drugs are related to the tumor cell proliferation, metastasis, and called apoptosis through in vitro cell experiments. Then, a initial exploration would be done on the effects on proliferation and metastasis of tumor and its possible mechanism by detecting the expression of Vascular endothelial growth factor (VEGF) and matrix metalloproteinases(MMPs). The results of this issue will be service in the follow-up studies and clinical treatment medication.Methods:To establish the model of HepG2and HUVEC cultured in vitro, preparat medicated serum by serum pharmacological method. Detecting the effect of the medicated serum of Qi-tonifying and Astringing drugs on the proliferation ability of HepG2and HUVEC by MTT method. Detecting the effect of the medicated serum of Qi-tonifying and Astringing drugs on the invasion and migration ability of HepG2and HUVEC by matrigel invasion assay. Detecting the protein and mRNA expression of VEGF, MMP-2and TIMP-2by Western Blot and Realtime-PCR technique.Results:Results of cell experiment show that:the medicated serum of Qi-tonifying and Astringing drugs in this issue have significantly inhibiting effect effect on the proliferation and migration ability of HepG2and HUVEC, compared with the blank serum contrast group, the difference has statistical difference (P＜0.05).Detecting the protein expression of MMP-2, TIMP-2, VEGF in HepG2by Western Blot technique, results of experiments show that:each medicated group compared with the blank serum contrast group, the protein expression of VEGF, MMP-2of each medicated group is reduced, the difference has statistical difference (P＜0.05or P＜0.01); the protein expression of TIMP-2of each medicated group is increased, the difference has statistical difference (P<0.05or P＜0.01).Detecting the mRNA expression of MMP-2, TIMP-2in HepG2by Realtime-PCR, results of experiments show that:each medicated group compared with the blank serum contrast group, the mRNA expression of MMP-2of each medicated group is reduced, the difference has statistical difference (P＜0.05or P<0.01); the mRNA expression of TIMP-2of each medicated group is increased, the difference has statistical difference (P<0.05or P<0.01).Detecting the protein expression of MMP-2, TIMP-2, VEGF in HUVEC by Western Blot technique, results of experiments show that:each medicated group compared with the blank serum contrast group, the protein expression of VEGF, MMP-2of each medicated group is reduced, the difference has statistical difference(P<0.05or P<0.01); the protein expression of TIMP-2of each medicated group is increased, the difference has statistical difference(P<0.05or P＜0.01).Detecting the mRNA expression of VEGF in HUVEC by Realtime-PCR, results of experiments show that:each medicated group compared with the blank serum contrast group, the mRNA expression of VEGF of each medicated group is reduced, the difference has statistical difference (P＜0.05or P<0.01).Conclusion:In vitro cell experiments can confirm that Qi-tonifying and Astringing drugs have inhibiting effect on cell proliferation and metastasis of HepG2, these above can be the mechanism of action of these drugs on Inhibition of proliferation and metastasis of tumor at the cellular level. The experimental results show that Qi-tonifying and Astringing drugs depress the protein and mRNA expression of MMP-2、TIMP-2、VEGF, these above can be the mechanism of action of these drugs on Inhibition of proliferation and metastasis of tumor at the molecular biology level.In addition, vitro cell experiments confirmed that Qi-tonifying and Astringing drugs have inhibition effect on cell proliferation and metastasis of HUVEC, these above can be the mechanism of action of these drugs on inhibiting angiogenesis at the cellular level. The experimental results show that Qi-tonifying and Astringing drugs depress the protein and mRNA expression of VEGF, these above can be the mechanism of action of these drugs on inhibiting angiogenesis of tumor at the molecular biology level.In summary, Preliminary showed that in vitro, Qi-tonifying and Astringing method have inhibition effect on proliferation and metastasis of tumor, which plays a double role in the anti-tumor and antiangiogenesis effect,, but the deeper mechanism needs to research and prove in vivo.