Anticancer Effect And Underlying Mechanism Of Natural Small Molecule Compounds In Colorectal Cancer And Glioma Cells

1 Effect and mechanism of small molecule compounds based on the energymetabolismof glioblastoma cellsPurpose:Gliomas are the most common type of intracranial tumors, more than half of primary brain tumor are gliomas. Despite recent advances of modern medicine, the average survival time of glioma pations don’t exceed 15 months, and it has long been observed that after surgical removal,treatment is almost never curative. The main way of tumor energy metabolism is aerobic glyco lysis, and this research is devoted to study the antitumor effects and mechanismsof ZZZ-1 through regulation of energy metabolism and fate in glioblastoma cells.Experiment design:In this study, we examined the effects of ZZZ-1 on proliferation and colony formationina series of glioma cell lines, also we tested the regulation of glucose and pyruvate uptake, lactate production, ATP production, oxygen consumption; The levels ofthe mitochondria membrane potential (ΔΨm) and reactive oxygen species (ROS) were examined, cellapoptosis and senesence were assessed; Additionally, the expression levels of apoptosis-related proteins, Bmi-1, GFAP andkey protease (PFKFB3 and LDH5) in glycolysis were also evaluated in gliomas. The ATP-synthase inhibitor oligmycin (OM) was used to explore the potential role of ZZZ-1. The anti-tumor activity, expression levels of VEGF, key protease (PFKFB3 and LDH5) in glycolysis, Bmi-1, GFAP, NeuN and Iba-1 were also evaluated in the U-87MG xenograft nude mice model in vivo.Result:1) Significant anti-glioma effect:ZZZ-1 significantly inhibitedcell growth and colony formationof human and rat glioma cells in a dose- and time-dependent manner;2) Cell death and differentiation was altered:ZZZ-1 induced lose of ΔΨm and generation of reactive oxygen species (ROS), inhibited the expression of PCNA and survivin, induced cell apoptosis in C6 cells and cell senescence in U-87MG cells, downregulated the expression ofBmi-land upregulated the expression of GFAP; 3) Transformation of energy metabolites in gliomas:treatment with ZZZ-1 for 24 h increased the glucose and pyruvate consumption and lactate production, intracellular ATP production and oxygen consumption were also increased after treatment with ZZZ-1 for different times,treatment with ZZZ-1 for 72 h suppressed the protein expression of PFKFB3 and LDH5; OM reduced the growth inhibition, the loss of ΔΨm, the overexpression of ROS, the increase of glucose consumption and ATP production inuced by ZZZ-1; on the other hand, ZZZ-1 could reduce the increased lactate production induced by OM; 4) Significant antiglioma effect in vivo:we used the nude mice U-87MG xenograft tumor model to study the antitumor effect of ZZZ-1 in vivo, and we found that ZZZ-1 inhibited human U-87MG xenograft growth and downregulated the expression of PCNA and survivin in vivo; The expression levels of key protease (PFKFB3 and LDH5) in glycolysiswere also decreased; Additionally, ZZZ-1 upregulated the expression levels of GFAP, NeuN and Iba-1 in vivo.Conclution:Collectively, these results indicate that ZZZ-1 plays important roles in modulating glioma cell proliferation and ZZZ-1 inhibits cell growth through modulating energy metabolic, inducing cell death and differentiation in glioma cells.2 Effect and mechanism of small molecule compoundsin Human Colorectal Cancer through modulation of multiple targetsPurpose:Colorectal cancer (CRC) is a global health problem with over one million new cases diagnosed and half a million deaths worldwide each year.This study is devoted to evaluate the antitumor effect, underlying mechanism and possible targets of caffeic acid 3,4-dihydroxyphenethyl ester (CADPE) on human CRC cells.Experimental Design:The effects of CADPE on cell growthand colony formation were examined in four CRC cell lines; The cellcycle progression, apoptosis and related protein expression were also analyzed in two CRC cell lines. Four MAPK inhibitors were used to explore the potential role of CADPE. The anticancer activity, induction of apoptosis and protein expression levels were also evaluated in human colorectal carcinoma xenografts in vivo. Effect of CADPE on human normal intestinal myofibroblast cell line was also evaluated. Results:We found that CADPE decreased the proliferation and colony formation of four different CRC cell lines, arrested cellcycle in G0/G1 phase and induced apoptosis in a subset of CRC cells, which were correlated with constitutive activation of p38, inhibition of nuclear factor kappa B (NF-κB) and its downstream targets such as signal transducer and activator of transcription 3 (STAT3) and c-Myc. CADPE also downregulated the cell survival (survivin) and cell cycle regulatory proteins (e.g., cyclin D1, CDK4, CDK6, p-Rb and E2F-1) in CRC cell lines. P38 inhibitor reversed the effectof CADPE on cell growth inhibition, cellcycle-related protein expression and apoptosis, but ERK and JNK inhibitors had no such effects. In vivo experiments, CADPE inhibited human colorectal cancer SW620 xenografts growth and induced apoptosis in immuno-deficient (SCID) mice, and decreased the protein expressionlevels of PCNA, survivin and cyclin D1 in tumor tissues. Interestingly, CADPE had a high level of security both in vitro and in vivo.Conclusion:These results indicated that CADPE plays important roles in modulating CRC cell proliferation and CADPE inhibits CRC cell growth by regulating multiple targets.