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1.
Figure 5

Figure 5. From: Inositol Hexaphosphate Suppresses Growth and Induces Apoptosis in Prostate Carcinoma Cells in Culture and Nude Mouse Xenograft: PI3K-Akt Pathway as Potential Target.

IP6 inhibits angiogenesis and decreases VEGF and eNOS expression in PC-3 tumor xenografts. Xenograft tissue samples from different treatment groups were subjected to IHC staining as detailed in ‘Materials and Methods’, analyzed qualitatively and quantitatively for the CD31-positive microvessels (A–D), and intensity of VEGF (E–H) and eNOS (I–L) immunostaining. In each case, the quantitative data shown are mean ± SEM of 5 tumor samples from individual mouse in each group. IP6, inositol hexaphosphate.

Mallikarjuna Gu, et al. Cancer Res. ;69(24):9465.
2.
Figure 2

Figure 2. From: Inositol Hexaphosphate Suppresses Growth and Induces Apoptosis in Prostate Carcinoma Cells in Culture and Nude Mouse Xenograft: PI3K-Akt Pathway as Potential Target.

IP6 decreases phosphorylation or expression of signaling molecules in PI3K/PDK1/Akt axis in PC-3 and C4-2B cells. After specific treatments, cells were collected at 6, 12 and 24 hrs, cell lysates prepared, and western blotting was done to analyze the effect of IP6 on the expression of (A) phosphorylated and total PI3K and PDK1, and (B) ILK1, phosphorylated and total Akt at Ser473 and Thr308, GSK3α/β (Ser9/21) and Cyclin D1 as detailed in Materials and Methods. IP6, inositol hexaphosphate.

Mallikarjuna Gu, et al. Cancer Res. ;69(24):9465.
3.
Figure 4

Figure 4. From: Inositol Hexaphosphate Suppresses Growth and Induces Apoptosis in Prostate Carcinoma Cells in Culture and Nude Mouse Xenograft: PI3K-Akt Pathway as Potential Target.

IP6 strongly decreases the expression of ILK1, pAkt (Ser473) and Cyclin D1 in PC-3 tumor xenografts. Xenograft tissue samples from different treatment groups were subjected to IHC staining as detailed in ‘Materials and Methods’, analyzed qualitatively and quantitatively for the expression of ILK1 (A–D), nuclear pAkt-Ser473 (E–H), and Cyclin D1 (I–L). In each case, the quantitative data shown are mean ± SEM of 5 tumor samples from individual mouse in each group. IP6, inositol hexaphosphate; NS, statistically not significant.

Mallikarjuna Gu, et al. Cancer Res. ;69(24):9465.
4.
Figure 6

Figure 6. From: Inositol Hexaphosphate Suppresses Growth and Induces Apoptosis in Prostate Carcinoma Cells in Culture and Nude Mouse Xenograft: PI3K-Akt Pathway as Potential Target.

IP6 strongly decreases HIF-1α DNA-binding activity in PC-3 tumor xenografts. Nuclear fractions were prepared from the tumor tissues and the DNA-binding reactions were done for 30 min in the dark as per the manufacturer’s instructions. The reaction mixture was resolved on 4–16% native Bis-Tris gel, run at 100 V for 60 min at room temperature in the dark and EMSA gel was analyzed and images were captured using the LI-COR Odyssey infrared laser imaging system. COS7 (CoCl2 treated) nuclear extract was used as positive control. Mutant HIF-1 IRDye end-labeled oligo was used to compete with the wild-type HIF-1 binding sequence along with the nuclear extract to establish the specificity of DNA-protein complex. The data shown are representative of three independent experiments. IP6, inositol hexaphosphate.

Mallikarjuna Gu, et al. Cancer Res. ;69(24):9465.
5.
Figure 1

Figure 1. From: Inositol Hexaphosphate Suppresses Growth and Induces Apoptosis in Prostate Carcinoma Cells in Culture and Nude Mouse Xenograft: PI3K-Akt Pathway as Potential Target.

IP6 inhibits growth and induces apoptosis in PC-3 cells. PC-3 cells were treated with 2 mM IP6 in complete media for 6, 12 and 24 hrs, and thereafter processed and analyzed as mentioned in ‘Materials and Methods’. A, total cell number and percentage of cell death was determined by Trypan blue dye exclusion assay. B, the percentage of apoptotic cell population was detected by ELISA Cell death assay. C, effect of IP6 on apoptotic markers, cleaved PARP and cleaved caspase-3, as determined by western blot analysis. In each case, membranes were simultaneously incubated with β-actin (loading control) and scanned with Odyssey Infrared Imager (84 µm resolution, 0 mm offset with medium or high quality). IP6, inositol hexaphosphate

Mallikarjuna Gu, et al. Cancer Res. ;69(24):9465.
6.
Figure 3

Figure 3. From: Inositol Hexaphosphate Suppresses Growth and Induces Apoptosis in Prostate Carcinoma Cells in Culture and Nude Mouse Xenograft: PI3K-Akt Pathway as Potential Target.

IP6 inhibits PC-3 tumor xenograft growth in nude mice together with in vivo anti-proliferative and pro-apoptotic effects. PC-3 cells were subcutaneously injected on the right flank of each mouse, 1% or 2% IP6 (w/v) was given in drinking water, and tumor volume was recorded as detailed in ‘Materials and Methods’. The average tumor volume is shown for the corresponding week (A), and tumor weight (g) per mouse (B) was noted at the end of the study. Control and respective IP6 treated groups were compared by one-way ANOVA followed by Bonferroni t-test for multiple comparisons. P<0.05 was considered statistically significant. C, tumor cell lysates were prepared and Western blot analysis was done as detailed in ‘Materials and Methods’ to assess the effect of IP6 on proliferation (PCNA) and apoptosis (cleaved PARP and cleaved caspase-3). IP6, inositol hexaphosphate; NS, statistically not significant.

Mallikarjuna Gu, et al. Cancer Res. ;69(24):9465.

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