Evaluation of the potential therapeutic effects of curcumin and lycopene in adult malignant brain tumour cells in vitro

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Abdullah Thani, Noor Azela;

Glioblastoma multiforme are the most challenging of cancers to treat and even with recent advances in therapeutic approaches, the prognosis following diagnosis remains poor. Novel therapeutic approaches able to target such tumours without resulting in significant toxici... View more
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    ........................................................................................................................................................... ..15 Figure 4: Chemical structures of curcumininoids; A) Curcumin, B) Demethoxycurcumin C) Bisdemethoxycurcumin and D) Cyclocurcumin

    ............................................................................................................................................................. 33 Figure 6: Metabolic pathways of curcumin in rodents and in vitro culture (rat and human hepatocytes). Taken from: Marczylo et al., 2007

    ............................................................................................................................................................. 39 Figure 13: A histogram shows a negative control in the flow cytometry analysis in which the primary antibody was omitted leaving only secondary antibody (with biotin conjugated) being incubated. M2 will represent the positive stained cells while M1 is negative in staining.

    ............................................................................................................................................................. 68 Figure 14: A quadrant in the Annexin V apoptosis. Each region in the quadrant i.e. lower right, lower left, upper right and upper left represents the population of cells. Axis with FL3-H indicates PI and axis with FL4-H indicates Annexin V.

    ............................................................................................................................................................. 71 Figure 15: The tubule network traced with Paint software (A and B) before analysed using Image J software (C). With the image J software, each colour represents a parameter measured in the angiogenesis. The four parameters are total length of the tubules, node and junction (red in blue circle) and branches (green)

    ............................................................................................................................................................. 74 Figure 16: Phase contrast micrographs of the normal brain cell line; A) CC2565-passage 10 and primary brain tumour cell cultures (GB biopsy-derived); (B) MUBS-passage 6 (C) MUBP-passage 9 (D) MUTC-passage 8 and (E) MUP-passage 8. The different morphology of cells was shown by the colour coded arrows which include spindle (red), small star (green), large star (blue) and broad, straight and blunt end (purple) in shapes. All cell cultures were viewed and compared under phase contrast microscopy (20X magnification)

    ............................................................................................................................................................. 77 Figure 21: Antigen expression of CC2565 cell culture following staining with different antibodies via immunocytochemistry. A) GFAP B) GD3 C) NG2 D) CD44 E) Beta-1 integrin. F) Negative control.

    ............................................................................................................................................................. 92 Figure 22: Antigen expression of MUBS cell culture following staining with different antibodies via immunocytochemistry. A) GFAP B) GD3 C) NG2 D) CD44 E) Beta-1 integrin. F) Negative control.

    ............................................................................................................................................................. 92 Figure 32: Four representative diagrams from Annexin V apoptosis assay of MUBP cells at different hour of incubation (following curcumin treatment); 24h (upper left box), 48h (upper right box), 72h (lower left box) and 96h (lower right box). In a diagram, each quadrant represents viable cells (lower left), early apoptosis cells (lower right), late apoptotic cells (upper right) and necrotic cells (upper left). Percentage of cell gated in the quadrant is made by CellQuest software ........................................................................................................................................................... 111 Figure 33: MUBP cells population from each quadrant (viable, early apoptotic, late apoptotic, necrotic) of every incubation time (following curcumin treatment) are represented in a bar graph.

    Error bars show standard error of mean. Data represents the mean ±standard error of mean for n=3 experiments with each experiment being carried out in duplicate. Percentage of cell gated was followed by statistical analysis (one way ANOVA-Fisher comparison) as to obtain the p value (*p value<0.05) and tabulated as below.

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