| Objective Microbial metabolites is an important source of modern drugdevelopment, currently the world’s top100drugs are being used, there are nearly50%of the drug derived from microorganisms, especially anti-cancer drugs, anti-infectives,cardiovascular disease and immune inhibitors, anti-cancer chemotherapy drugs,antimetabolites, and heavy metals in addition, the antibiotic is most important, most ofthese drugs are occasionally discovered and utilized, and further processing of themicrobial metabolism in the course of the study products, in medicine source of bacteriato develop more actinomycetes, fungi, Streptomyces and Bacillus, it is preciselybecause of the high degree of development, so even consume a lot of time to spend a lotof money, it is difficult to isolate from these microorganisms new effective drugs. Sincethe fifties of the last century, scientists have observed that the bacteria secrete stickyproduct has biological activity of antibiotics, which has a species-specific (differentstrains of the same category metabolites different, different types of bacteria can producea number of similar structure metabolites) characteristics, making sticky bacteria hasbeen favored more and more researchers, and in2008by the U.S. FDA approval for thelisting of epothilone (epothilones) is the most famous extracted from the sticky anti-bacterial metabolites in tumor substances, anti-cancer drugs has become the first line,because it has a similar pro-taxol microtubule polymerization activity, with the multi-drug-resistant tumor cells to strong toxicity, have less side effects because no complexchemical structure, good water solubility and pathways from microbial fermentation, isconsidered paclitaxel replacement products, better anti-cancer drug, is one of the mostimportant value sticky bacteria can produce a rich variety of secondary metabolites theproduct, with a variety of new structures, innovative and diverse characteristics ofmechanism of action, is recognized as an important source of outside exceptactinomycetes new kinds of microbes drugs Qinba mountain flora and fauna as one of China’s largest natural resource protection areas, microbial reserves in the country andeven the world are unique, which contains the sticky type of bacteria is so diversemetabolites were screened to find anticancer drugs, has unique advantages and greatpotential, sticky bacteria commonly found in soil and decaying trees, vegetation, andcan ferment the extremely rich secondary metabolites present from sticky bacterialmetabolites unearthed hundreds of biologically active ingredients, yet the vast majorityof the microbes to discover the other group, metabolites which have a higher proportionof anti-tumor substances, therefore, screening sticky bacterial secondary metaboliteslooking for anti-cancer drugs, has unique advantages and great potential. Theexperimental study was to investigate the role of sticky bacteria obtained from twosecondary metabolites Qinba Mountain (NX52and NX83) HT-29cell proliferation afterits generated metabolite explore the possible mechanism of action.Method Develop stable, healthy passaged HT-29colon cancer cell lines, cellswere observed by inverted optical microscope morphology, growth curve by MTTmethod of HT-29cells, and colleagues estimated the population doubling time, applydifferent MTT assay concentration (0.1mg/ml,1.0mg/ml,10mg/ml) of the two stickybacterial metabolites (NX52, NX83) HT-29colon cancer cell proliferation of culturedhuman and metabolites combined with an inverted microscope viewing morphology ofHT-29cells after interaction.Result Successfully cultivate a stable, healthy passaged HT-29colon cancer celllines, single non-adherent cells when oval-shaped cells adherent monolayer growth,elongated strip of irregular polygons, faster cell growth between clusters of cells, whenthe cells covered the bottom80%or more cells into a group gathered adhesion, cellaggregation dense, clear boundary, clear cytoplasm, plump, good transparency, celldoubling time of40hours, sticky bacterial metabolites in each dosing group comparedwith the control group reduced proliferation in a dose-dependent effect (0.1mg/ml,1mg/ml and10mg/ml) and time-dependent effect (24h,48h and72h), secondarymetabolites through the treated HT-29cell proliferation and morphological changesoccurred, the degree of change with time of action and concentration of the productincreased metabolism increases the number of inverted microscopic holes dosing hole significantly reduced compared with the control cells small, significantly increasedspacing between cells, cell shrinkage, blurred edges, vacuoles, irregularly shaped cellsappeared in most of the thin cytoplasm, shapes, visible part of the cell fragmentation.Conclusion Train a stable passage of HT-29cell line can be used as a good modelfor in vitro experimental cancer research, NX52, NX83are secondary metabolitesisolated bacteria from sticky bred in Qinba Mountain, both on colon cancer HT-29cellswas inhibited, the primary metabolite extracts from a variety of substances, from anti-metabolite mechanism to consider the characteristics of sticky bacteria and epothilonetumors, reason to believe that the two metabolites of this experiment may contain anti-tumor substances, the mechanism may be similar with the epothilone, such as thepromotion of microtubule structure and stability of cells and thus play stop mitosis orinhibit electron transport, cytoskeleton damage, inhibiting nucleic acid polymeraseactivity and the like affect cell metabolism, cell destruction structure, inhibition of DNAreplication to achieve anti-tumor effect, In summary, there may have colon cancer HT-29cells was inhibited bioactive substances secondary metabolites of bacteria in thisexperiment, two sticky, sticky for the further study to determine the bacterial secondarymetabolites have anti-tumor effects the separation and purification of the monomersubstances and bacteriostatic substances or development of anticancer drugs to provideexperimental basis. |
PDF Full Text Request