G0/G1 arrest and S phase inhibition of human cancer cell lines by inositol hexaphosphate (IP6).
ABSTRACT Inositol hexaphosphate (InsP6 or IP6) has shown a striking anti-cancer activity in both in vivo and in vitro models. In an attempt to elucidate the mechanism(s) underlying the anti-neoplastic potential of IP6, we investigated its effect on cell cycle progression of MCF-7 estrogen receptor (ER)-positive and MDA-MB 231 ER-negative human breast cancer cell lines and HT-29 human colon cancer cells.
The anti-proliferative effect of IP6 was evaluated using dual-parameter flow cytometric measurements of DNA content, versus the incorporation of 5-bromo-2-deoxyuridine (BrdU) to determine cells actively synthesizing DNA. Combined analysis of the expression of cell cycle-related proteins, proliferation marker Ki-67 and proliferating cell nuclear antigen (PCNA) versus DNA content were used to determine the amount of proliferating cells in each phase, engaged in cell cycle transit.
After 3 days of treatment with 5 mM IP6, S-phase, as estimated by BrdU uptake, was significantly decreased in all three cell lines (p = 0.002). MCF-7 and HT-29 cells accumulated in the G0/G1 range of DNA contents (p = 0.002 and p = 0.001, respectively). MDA MB-231 cells transiently accumulated in G0/G1 only after 2 days (p = 0.01). There was a significant decrease in the percentage of Ki-67 expression in IP6-treated cells, from 82.8+/-3.0% to 66.8+/-4.2% in MCF-7 (p = 0.007), from 93.4+/-4.6% to 71.7+/-3.3% in MDA-MB 231 (p = 0.004), and from 95.2+/-1.2% to 73.5+/-2.5% in HT-29 cells (p = 0.002) respectively. PCNA expression levels were also significantly decreased by IP6 in all three cell lines (MCF-7 p = 0.0007; MDA-MB 231 p = 0.0006; HT-29 p = 0.0001).
These results show that IP6 controls the progression of cells through the cycle by decreasing S- phase and arresting cells in the G0/G1-phase of the cell cycle. A significant decrease in the expression of proliferation markers indicated that IP6 disengaged cells from actively cycling. Further investigations of cell cycle regulators may lead us to a better understanding of the mechanism(s) of the anti-neoplastic action of IP6.
Full-textDOI: · Available from: A.K. M Shamsuddin, May 08, 2014
- SourceAvailable from: Hae-Jeung Lee[Show abstract] [Hide abstract]
ABSTRACT: Chemoprevention is considered a rational strategy for dietary approaches to prevention of cancer. Multiple lines of evidence suggest that many of our dietary principles are able to intervene in the multistage carcinogenesis process and phytic acid (inositol hexaphosphate, IP6), a phytochemical present in a variety of plant species, has been shown to prevent various cancers, including those of the mammary gland, colon and liver. However, the mechanism of chemoprevention by IP6 has not been fully elucidated. In the present study, we examined the effects of inositol and/or IP6 supplementation on rat hepatocarcinogenesis initiated by diethylnitrosamine (DEN) and promoted by partial hepatectomy (PH). Supplementation with either inositol or IP6, or their combination, starting one week prior to administration of DEN, resulted in a significant decrease in both the area and the number of placental glutathione S-transferase positive (GST-P+) foci, a preneoplastic marker for DEN-initiated hepatocarcinogenesis. The administration of inositol and/or IP6 in drinking water caused marked enhancement in the glutathione S-transferase (GST) activity. In addition, the production of thiobarbituric acid reactive substances and the catalase activity were significantly reduced in rats supplemented with inositol and /or IP6. Based on these findings, it is likely that the chemopreventive effects of inositol and/or IP6 on rat hepatocarcinogenesis initiated by DEN and promoted by PH are associated with induction of GST activity and suppression of lipid peroxidation.Asian Pacific journal of cancer prevention: APJCP 6(1):41-7. · 2.51 Impact Factor
Article: Anti‐cancer function of phytic acid[Show abstract] [Hide abstract]
ABSTRACT: Summary Inositol hexaphosphate (InsP6 a.k.a. phytic acid or IP6) is ubiquitous. In the plant kingdom it is particularly abundant in cereals and legumes; in much smaller amounts IP6 and its lower phosphorylated forms (IP1−5) are contained in most mammalian cells, where they are important in regulating vital cellular functions. Both in vivo and in vitro experiments have demonstrated striking anticancer (preventive as well as therapeutic) effects of IP6. Inositol also is anti-carcinogenic, albeit to a lesser extent; it acts synergistically IP6 in inhibiting cancer. In addition to reduction in cell proliferation, IP6 increases differentiation of malignant cells often resulting in reversion to the normal phenotype. IP6 is quickly absorbed from the rat stomach and upper intestine and distributed as inositol and IP1. In vitro, it is instantaneously taken up by malignant cells undergoing variable dephosphorylation to inositol and IP1−5, pointing towards their role in mediating the action of IP6. In humans, IP6 has recently been detected in urine, plasma and other biological fluids; the levels fluctuating with ingestion or deprivation of IP6 or IP6-rich diet. As IP6 is high in high-fibre diets, these also may explain, at least in part, the epidemiological observation showing the association of ingesting high-fibre diets with a lower incidence of certain cancers. Along with safety, the reproducible efficacy of IP6 and inositol in the prevention of cancer in laboratory animals warrant their inclusion in our strategies for cancer prevention and perhaps therapy in humans. Aside from the anticancer action, IP6 and inositol also have numerous other health benefits. All these facts of normal physiological presence of IP6 in our body the level of which fluctuates with intake, association of an IP6-rich diet with low incidence of several diseases and vice versa, and finally reversal of some of these conditions, at least in part, by IP6 supplementation strongly argue in favour of its inclusion as an essential nutrient or perhaps a vitamin.International Journal of Food Science & Technology 09/2002; 37(7):769 - 782. DOI:10.1046/j.1365-2621.2002.00620.x · 1.35 Impact Factor
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ABSTRACT: Phytic acid or IP6 has been extensively studied in animals and is being promoted as an anti-cancer agent in health food stores. It is naturally found in legumes, wheat bran, and soy foods. It is believed to be the active ingredient that gives these substances their cancer fighting abilities. Proposed mechanisms of action include gene alteration, enhanced immunity, and anti-oxidant properties. A Medline search from 1966 to May 2002 using the keywords phytic acid and cancer, and limiting the search to the subheadings of therapeutic uses, prevention, and adverse effects revealed 28 studies. These studies were included in the review. A great majority of the studies were done in animals and showed that phytic acid had anti-neoplastic properties in breast, colon, liver, leukemia, prostate, sarcomas, and skin cancer. There were no human studies. Side effects included chelation of multivalent cations, and an increase in bladder and renal papillomas. This increase in papilloma formation only occurred with the sodium salt of phytic acid. It did not occur with either the potassium or magnesium salts. There is a large body of animal evidence to show that phytic acid may have a role in both the prevention and treatment of many forms of cancer. There is clearly enough evidence to justify the initiation of Phase I and Phase II clinical trials in humans.Complementary Therapies in Medicine 01/2003; 10(4):229-34. DOI:10.1016/S0965-2299(02)00092-4 · 2.22 Impact Factor