Drug Serum Of Sijunzi Decoction Impact On Biological Characteristics Of Side Population Cells In Human Gastric Carcinoma Cell Lines BGC-823、SGC-7901

Part I Isolation of side Population cells in human gastric carcinoma cell lines BGC-823,. SGC-7901Objective:Detection and isolation the proportion of SP cell in gastric carcinoma cell lines BGC-823、SGC-7901.Methods:Choice low、moderately degree of differentiation of gastric carcinoma cell lines SGC-7901、BGC-823,stained by fluorochrome Hoechst33342, controlled with the Presence of verapamil, SP cells Were analyzed and isolated by fluorescence-activated cell sorting(FACS). Detection and isolation the proportion of SP cell from human gastric carcinoma cell lines.Results:SP cell was detected in gastric carcinoma cell line BGC-823with the proportion of2.60%±1.25%, gastric carcinoma cell line SGC-7901with the proportion of2.53%±0.85%.Conclusion:There existing a certain number of SP cells in human gastric carcinoma cell lines SGC-7901, BGC-823, which can be isolation. There are some differences the proportion of SP cells in various of differentiation degree of tumor cells.Part II Identify the biological characteristic of side population cells in human gastric carcinoma cell lines BGC-823、SGC-7901Objective:Appraisal biology characteristics of the SP cells isolation from gastric carcinoma cell lines BGC-823、SGC-7901through proliferation activity in vitro, matrigel invasion assay, tumorigenicity in vivo preliminary, to investigate the feasibility of starting Research on gastric cancer stem cell from the side population.Methods:Compare the proliferation activity in vitro of the SP、NSP cells and the total cells by cell growth curve (CCK-8), and colony formation assay using matrigel invasion assay. Compare the differences of invasive ability in vitro of the SP、NSP cells and the total cells using xenograft transplantation of NOD/SCID mouse tumor experim ents. Comparing SP and NSP cells and the total cells with sorting tumorigenicity differences in vivo. Identified the characteristic of differentiation through the tumor in vivo differentiation phenomenon.Results:Cell growth curve shows that the proliferation ability of BGC-823and SGC-7901SP cell is obviously higher than NSP cell and the total cell (P<0.05). Flat cloning experiment results shows that the clone formation rate of BGC-823and SGC-7901SP cell is greater than NSP cell and the total cell (P<0.05). Transwell cells invasion assay showed that BGC-823and SGC-7901SP cell invasive ability less than NSP cell and the total cell (P<0.05). Xenograft tumor experimental results show that BGC-823and SGC-7901SP cell showed stronger tumorigenicity than NSP cell and the total cell. SP and NSP cell and the total cell tumor in vivo, all can be transformed to different differentiation degree of gastric carcinoma cells.Conclusion:People gastric carcinoma cell lines BGC-823、SGC-7901SP cell has a strong proliferation in vitro and tumorigenicity than NSP cell and the total cell without sorting, invasive ability is weak, has the multidirectional differentiation characteristics, properties in line with the cancer stem cell, the SP cell enrich cancer stem cells, can be used as starting Research on gastric cancer stem cell.PartⅢ Drug serum of sijunzi decoction impact on biological characteristics of side population cells in human gastric carcinoma cell lines BGC-823、SGC-7901Objective:The intervention drug serum of sijunzi decoction impact on biological characteristics of SP cells in human gastric carcinoma cell lines through proliferation activity in vivo and tumorigenicity research, which is provided objective experimental and theoretical basis for traditional Chinese medicine in the treatment of gastric cancerMethods:Make animal drug serum, high performance liquid chromatography (HPLC) detect the concentrations of serum drug, Compare the impaction on proliferation activity in vitro from cell growth curve (CCK8) and colony formation assay after intervention of drug serum, application xenograft transplantation of NOD/SCID mouse tumor experiments Compare the impact on tumorigenicity in vivo after intervention of drug serum.Results:High performance liquid chromatography (HPLC) analysis shows that drug active components which are existed in serum, cell growth curve showed that BGC-823and SGC-7901SP and NSP cells after the intervention of drug serum proliferation activity below without which is the intervention (P<0.05), xenograft transplantation experiment results showed that BGC-823and SGC-7901SP and NSP cells after the intervention of drug serum showed weak tumorigenieity in vivo than the one without intervention by tumors growth curve (P<0.05).Conclusion:The intervention of drug serum Sijunzi decoction in gastric cancer cell lines BGC-823and SGC-7901SP and NSP cells decreased in proliferation activity in vivo and tumorigenicity, drug serum of sijunzi decoction can inhibite proliferation activity and tumorigenicity of SP cells on gastric cancer BGC-823and SGC-7901.