The Proliferation Of Radix Salviae Miltiorrhiae. And Agrimonia Pilosa Ledeb. On A549 Cells And The Influence On Expression Of Angiogenesis-related Genes

Cancer patients have poor survival.For cancer treatment,clinical chemotherapy,surgery is the main treatment,which has significant effect.But drawback is that these treatments have more side effects.So looking for both anti-tumor cell and relatively small side effects drug and treatment particularly important.Traditional Chinese medicine has a long history of clinical application.Anti-tumor effect of Chinese medicine has many advantages.therefore,anti-tumor effect of Chinese medicine has become popular.In this study,to explore the effects of Radix Salviae Miltiorrhiae.and Agrimonia pilosa Ledeb.water extract on proliferation and apoptosis of human lung cancer A549 cell line;To study Chinese medicine which active blood circulation and remove blood stasis whether has the effect of the promotion of tumor metastasis or not. Experimental section includes four parts:1.MTT assay was used to observe the anti-proliferation effect of six Chinese medicine water extract on A549 cells for 48h.2.MTT assay was used to observe the anti-proliferation effectof Radix Salviae Miltiorrhiae. water extract on A549 cells for 24h,48h,72h;Morphological changes were observed under optical microscopy;AO/EB fluorescence double-dye technology was performed to observe the apoptosis of cells treated for 48h in fluorescence microscope;The apoptosis rates of A549 cells treated by Radix Salviae Miltiorrhiae.water extract for 24h were determined by flow cytometry (FCM).3.MTT assay was used to observe the anti-proliferation effect of Agrimonia pilosa Ledeb. water extract on A549 cells for 24h,48h,72h;Morphological changes were observed under optical microscopy;AO/EB fluorescence double-dye technology was performed to observe the apoptosis of cells treated for 48h in fluorescence microscope;The apoptosis rates of A549 cells treated by Agrimonia pilosa Ledeb.water extract for 24h were determined by flow cytometry (FCM).4. Tow Genes(COX-2,VEGF),which are enhance angiogenesis factor,were chosen to be validated by Q-PCR. The expressions of COX-2,VEGF Genes were detected by Q-PCR at 12h after 5mg/ml,2.5mg/ml,1.25mg/ml Radix Salviae Miltiorrhiae.and Agrimonia pilosa Ledeb. water extract treated A549 cells.Results:1.Radix Salviae Miltiorrhiae.and Agrimonia pilosa Ledeb. water extract has the best proliferation of A549 cells.2.Radix Salviae Miltiorrhiae.water extract inhibited the proliferation of A549 cells in a dose-dependent manner,and the best proliferation was after being treated for 48h;Morphological changes were dramatical under optical microscopy;In experiment group appeared yellow or chrysoidine fluorescence,cont racted,beaded and crescent and the number of apoptotic cells increased;FCM analysis showed that cells were induced apoptosis dose-dependently.3.Agrimonia pilosa Ledeb.water extract inhibited the proliferation of A549 cells in a dose-dependent manner.And the best proliferation was after being treated for48h; Morphological changes were dramatical under optical micrscopy;In experiment group appeared yellow or chrysoidine fluorescence,cont racted,beaded and crescent and the number of apoptotic cells increased;FCM analysis showed that cells were induced apoptosis dose-dependently.4. Expression of COX-2 and VEGF Genes detected by Q-PCR:The expression level of COX-2 and VEGF after 5mg/ml,2.5mg/ml,1.25mg/ml Radix Salviae Miltiorrhiae. on A549 was lower than that in blanke control groupe and in a dose-dependent manner.The expression level of COX-2 after A549 cells induced by 5mg/ml,2.5mg/ml,1.25mg/ml Agrimonia pilosa Ledeb.was lower than that in blanke control groupe and was not in a dose-dependent manner.But the expression level of VEGF after A549 cells induced by 5mg/ml,2.5mg/ml,1.25mg/ml Agrimonia pilosa Ledeb.was in a dose-dependent manner. This shows that Radix Salviae Miltiorrhiae.and Agrimonia pilosa Ledeb.water extract could indirectly inhibit tumor cell metastasis.