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EXPERIMENTAL PROCEDURES

These mice were divided into 2 feeding groups: a 20 mice on a sterile, a chemically-defined AINbased diet containing 0. Body weights and average diet intake for each group were measured weekly. Plasma samples were analyzed for amino acid content, insulin, IGF-1, FGF21, leptin, adiponectin, cholesterol, triglycerides, and glucose.

Tumor volumes were determined using the formula: 0. Histopathology analysis was performed by Dr Timothy Cooper Penn State College of Medicine , and the percent of lesions morphologically resembling mild, moderate, or florid intraductal papillomas IDPs ; ductal carcinoma in situ DCIS ; or invasive carcinoma were calculated. The tumor sections also were used for immunohistochemical analysis of proliferation and apoptosis for all groups.

Additionally, the mammary glands from 10 mice of each group were fixed in NBF for histology. At sacrifice, organ weights for liver, kidneys, spleen, mammary fat pads, and left gastrocnemius muscle were recorded. The percentages of Ki67 and cleaved caspase-3 positive cells counted in triplicate under a high power field X were used as a measure of proliferation and apoptosis, respectively. Staining was completed using a Dako Autostainer Plus.

Beverly, MA. Blood glucose was measured using an AbbottH Freestyle glucometer and glucose test strips. Four experiments with 6 replicates per treatment were conducted. Ten micron sections were obtained from the formalin-fixed, paraffin-embedded MCF10AT1 tumors and mammary fat pads of the athymic nude mice. This data set was screened for those patients containing data on mRNA levels from Agilent microarrays.

Histological analyses of mammary and tumor sections from the nude mice were analyzed using an unpaired Mann-Whitney one-tailed t-test to determine significance. Physiological parameters of mice on CF and MR were examined. Muscle mass decreased in the mice on MR diet. Future studies may provide a more in depth understanding of the changes that were observed in the muscle. One of the hallmarks of cancer is increased metabolic activity of the cells. Decreased plasma amino acid levels in cancer patients supports the concept that the tumor is parasitizing the organism and altering protein metabolism that would then affect the amino acid concentrations [ 28 ].

Equally important is the finding that the sulfur-containing amino acids methionine, cysteine, and taurine were reduced significantly in plasma, providing clear evidence that the MR diet reduces sulfur-containing amino acids throughout the animal. As predicted from previous studies of mice on a MR diet [ 11 , 29 ], reductions in plasma triglycerides, IGF1, and glucose were observed Fig.

Cholesterol levels were slightly elevated in the MR mice. Leptin was not significantly changed Fig. These differences in leptin levels may be due to strain differences between the two studies. Selected analytes measured in the plasma from nude mice on MR or CF diet. Tumor weights averaged The mice on the MR diet had a notable decrease in tumor weight to This suggests that lesions formed prior to the initiation of the MR diet were not eliminated by MR, but that MR slowed tumor progression and reduced the size of tumors formed by the implanted MCF10AT1 cells.

The fact that there was a population of mice that acquired an invasive carcinoma phenotype suggests that some cells may be able to escape the MR inhibition through an unknown mechanism. Left mammary gland 4 is indicated with arrows. To understand a possible mechanism for the decreased tumor size in the mice on the MR diet, proliferation and apoptosis were examined. Histological sections of the both tumor and nearby mammary tissue were stained for Ki67, a marker for proliferation.

A total of 20 animals per treatment were examined for levels of Ki67 staining. Representative images demonstrate that the number of Kipositive stained cells was reduced in the tissue from mice on the MR diet Fig. The number of proliferating cells in the tumor tissue was Consecutive sections of tumor tissue were stained for activated caspase-3, as an indicator of apoptosis Fig. Increases in apoptosis and decreases in proliferation occur in mice on the MR diet when compared to mice on the CF diet.

Mann-Whitney unpaired one-tailed t-test was performed on the samples. The decrease in proliferation combined with the increase in apoptosis suggests that MR may perturb the cell cycle. Expression of murine cell cycle inhibitors P21 and P27 and the cell cycle regulator cyclin D1 were measured by real-time PCR in the nude mice. Although there was a slight decrease in cyclin D1, this effect was not significant Fig. P21 expression was significantly increased Fig. To determine whether the human cell-derived tumors were affected similarly by MR, paraffin sections of embedded tumors from CF or MR mice were collected, and RNA was isolated.

There was a trend indicating that human P21 was increased in the tumors from MR fed mice Fig. Methionine restriction increases cell cycle inhibitors P21 and P27, while decreasing cyclin D1. Mann-Whitney unpaired one-tailed t-test. The next day, media was changed to either regular media This suggests that the MR effect on the cell cycle inhibitors may be a similar response to MR in both breast cancer cells and immortalized breast cells.

The differences in proliferation may indicate that MR may be more effective at hindering invasive breast cancer cells. In both cell types, time had significant affect on proliferation. This data set contains patients, of which data are available for patients regarding mRNA expression levels from Agilent microarrays.

Four patients had decreases in both P21 and P27, 11 patients had decreases in P21, and 14 patients had decreases in P Of the 52 patients, information was available regarding human epidermal growth factor receptor 2 Her2 , estrogen receptor ER , and progesterone receptor PR status in 32 patients. This may suggest that cancer patients following an MR diet may benefit, since increased levels in P21 and P27 can inhibit the cancer cell proliferation, providing a longer period of time for other established cancer therapies to be effective.

Survival curve of patients with decreases in cdkn1a or cdkn1b gene expression compared to the breast invasive carcinoma patients with mRNA Agilent microarray data. A rapid decrease in overall survival occurs in patients with decreases in cdnk1a or cdnk1b gene expression when compared to all patients with cancer. Dietary MR has been identified as a strategy for disease prevention and increased lifespan in experimental animals [ 10 , 19 , 31 , 32 ].

Our findings support this hypothesis and indicate that MR inhibits the growth of breast cancer tumors and induces apoptosis. High levels of IGF1 and insulin in humans have been linked to increased risk of breast cancer [ 33 ]. In carcinogenesis, growth factors, such as IGF1 and insulin, stimulate growth and progression of cancer [ 34 ]. Methionine functions as the donor for the C2-C4 and amine nitrogen during the synthesis of the polyamines spermidine and spermine. Ornithine, the precursor of putrescine which in turn is the precursor of spermidine and spermine , is increased in mice on the MR diet.

Polyamines are involved in the regulation of proliferation, growth, and survival of cells [ 35 ]. High levels of polyamines have been identified in several cancers [ 36 ], and the inhibition of polyamine synthesis has been shown to have antitumor effects on skin, colon, and prostate cancers [ 37 , 38 ].

The increase in ornithine suggests an inhibition of polyamine production. The metabolism of polyamines is strictly controlled and contains two rate-limiting enzymes: ornithine decarboxylase and S-adenosyl methionine decarboxylase [ 39 ]. Ornithine, spermidine, and spermine were previously reported to be increased in the liver of MR rats [ 29 ].

In the Perrone et al. The conflicting findings regarding polyamine synthesis suggest that either MR effects on the tumor are independent of polyamine synthesis, or alternatively, polyamine function and synthesis are regulated differently in liver and mammary gland. Taurine is a sulfur-containing amino acid that contributes to cell volume homeostasis and affects apoptosis mechanisms [ 40 ].

Taurine plasma levels are decreased in mice on MR. In cancer, volume-sensitive taurine correlates with cisplatin resistance [ 41 ]. Recently, taurine has been shown to induce apoptosis through PUMA p up regulated modulator of apoptosis in human colorectal, ovarian, and cervical cancer cells [ 41 — 43 ]. A decrease in taurine levels resulted in reduced cell volume that induced levels of activated caspase-3, which led to apoptosis in cervical adenocarcinoma cells [ 44 ]. Taurine has been examined as a novel tumor marker in female breast cancer patients.

A decrease in serum taurine levels was observed in people with breast cancer or at high risk of breast cancer. Further, an inverse correlation between vascular endothelial growth factor VEGF,marker for angiogenesis and taurine concentrations has been demonstrated [ 45 ]. Therefore, the connection between taurine, MR, and cancer is convoluted and complicated, and further research is needed to understand the implication of reduced plasma levels of taurine in MR mice. In both the xenograft model and in breast cancer cell lines, the mechanism by which MR inhibits tumor progression appears to be a coordinated effort of inhibiting the cell cycle by stimulating the cell cycle inhibitors, P21 and P Levels of P27 were not significantly changed in the xenograft model, suggesting that the effect of MR is more related directly to inhibiting P21 than P Additionally, P27 also was elevated in the breast cancer cell lines, suggesting that it too may be involved in the suppression of tumor progression.

Survival analysis revealed that patients with decreased P21 or P27 expression had reduced survival. The results from the present study indicate that MR may be a protective agent against cancer progression, but does not completely inhibit cancer progression. Therefore, the application of MR in a clinical setting could be both safe and a feasible option for arresting the progression of breast cancer while providing patients with more time to be treated by conventional methods to eradicate the cancer.

Further studies are needed to examine the effect of MR in combination with chemotherapeutic and immunotherapeutic treatments. Patient-derived breast tumor xenografts facilitating personalized cancer therapy. Breast Cancer Res. Breast and cervical cancer in countries between and a systematic analysis. Regulation of pyruvate metabolism in metabolic-related diseases.

Rev Endocr Metab Disord. Warburg O. On the origin of cancer cells. Dang CV. Glutaminolysis: supplying carbon or nitrogen or both for cancer cells? Cell Cycle. Omodei D, Fontana L. Calorie restriction and prevention of age-associated chronic disease. FEBS Lett. Weindruch R. Dietary restriction, tumors, and aging in rodents. J Gerontol. J Nutr. Lifespan-extending caloric restriction or mTOR inhibition impair adaptive immunity of old mice by distinct mechanisms.

Aging Cell. Methionine-deficient diet extends mouse lifespan, slows immune and lens aging, alters glucose, T4, IGF-I and insulin levels, and increases hepatocyte MIF levels and stress resistance. PLoS One. Methionine restriction prevents the progression of hepatic steatosis in leptin-deficient obese mice. Metab Clin Exp. Cysteine supplementation reverses methionine restriction effects on rat adiposity: significance of stearoyl-coenzyme A desaturase. J Lipid Res. Metabolic adaptations to methionine restriction that benefit health and lifespan in rodents.

Exp Gerontol. Methionine restriction effects on mitochondrial biogenesis and aerobic capacity in white adipose tissue, liver, and skeletal muscle of F rats. Sanchez-Roman I, Barja G. Regulation of longevity and oxidative stress by nutritional interventions: role of methionine restriction.

J Bioenerg Biomembr. Low methionine ingestion by rats extends life span. Steinmann, L. Boardman, S. Thibodeau, and R. One-step fluorescent probe product-enhanced reverse transcriptase assay. Arnold, R.


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Hepler, and P. Update by B. Flow Cytometric quantification of surface-displayed recombinant receptors on staphylococci. Goetsch, C. Samuelson, T. Nguyen, A. Robert, M. Binz, and S. Update by C. Jacobsson and L. Frykberg Update by K. Biopanning phage display libraries using magnetic beads Vs.

The Cell Cycle (and cancer) [Updated]

Polystyrene Plates. McConnell, T. Dinh, M. Le, and D. Fusion proteins could generale false positives in peptide phage display. Murthy, I. Ekiel, S. Shen, and D. Update by K. Phosphorylation-Directed antibodies in high-flux screens for compounds that modulate signal transduction. Alberta and C. Update by J. Two-Hybrid selection. Mammalian Two-Hybrid system: A complementary approach to the yeast two-hybrid system. Luo, A. Batalao, H. Zhou, and L. Development of a yeast trihybrid screen using stable yeast strains and regulated protein expression.

Fuller, M. Morse, J. White, S. Dowell, and M. Use of a dicistronic expression cassette enconding the green fluorescent protein for the screening and selection of cells expressing inducible gene products. Mosser, A. Caron, L.

Arrest Signaling

Bourget, P. Jocolieur, and B. Update by D. Dual-function reporte protein for analysis of gene expression in living cells. Day, M. Kawecki and D. Fusion of green fluorescent protein with the Zeocin Resistance marker allows visual screening and drug selection of transfected euraryotic cells.

Bennett, C. Cox, J. Valdez, L. Perlaky, Z. Cai, D. Henning, and H. Fission yeast expression vectors adapted for positive identification of gene insertion and green fluorescent protein fusion. Zhao, R. Elder, M. Chen, and J. Update by Y. Tracking and quantitation of retroviral-mediated transfer using a completely humanized, red-shifted green fluorescent protein gene.

Muldoon, J. Levy, S. Kain, P. Kitts, and C. Link Jr.

Does the ribosome translate cancer? | Nature Reviews Cancer

A combined selection and reporter gene for retroviral and transgenic studies. Blake, P. Salinas, and S. Retroviral gene transfer in chondrogenic limb bud micromass cultures. Stott, Y. Lee, and C. Update by W. Wang, N. Kasahara, and C. High-efficiency gene transfer and pharmacologic selection of genetically engineered human keratinocytes. Deng, K. Choate, Q. Lin, and P. Expression genetics : differential display Book 6 editions published in in English and held by 51 WorldCat member libraries worldwide.

Differential display methods and protocols by Peng Liang Book 8 editions published in in English and held by 25 WorldCat member libraries worldwide. Abstracts of papers presented at the Ninth Cold Spring Harbor Conference on Cell Proliferation : growth of cells in hormonally defined media, September 1-September 6, Book 3 editions published in in English and held by 16 WorldCat member libraries worldwide. Cell Cycle and Growth Control : Biomolecular Regulation and Cancer by Stein 1 edition published in in English and held by 5 WorldCat member libraries worldwide The groups of specialized cells that make up the various human tissues depend on an intricate communication network to regulate gene expression that in turn mediates growth, cell--type specific function, division, and programmed cell death.

This network consists of extracellular signals interacting with the receptors of individual cells and determining the fate of each. Since this regulatory system plays a critical role in complex tissue, aberrations or malfunctions often accompany the onset and progression of cancer.

Cell Cycle and Growth Control: Biomolecular Regulation and Cancer, Second Edition provides a solid basis for understanding cell cycle and growth control as it relates to biological regulation, with a special emphasis on examining these processes in the context of cancer. The only book to comprehensively cover both the foundations and cutting--edge advances in understanding cell cycle and growth control, this text also contains an expert perspective on innovative strategies for cancer treatment, making it a vital companion for researcher and clinician alike.

Ruth Sager : February 7, March 29, Book 1 edition published in in English and held by 3 WorldCat member libraries worldwide. The cell cycle in cancer prognosis by William A Creasey Book 1 edition published in in English and held by 3 WorldCat member libraries worldwide.

Growth of cells in hormonally defined media : conference ; Cold Spring Harbor - N.

Pardee, Arthur B. (Arthur Beck) 1921-

Cancer : fundamental ideas by Arthur B Pardee Book 1 edition published in in English and held by 2 WorldCat member libraries worldwide. The cell cycle in cancer prognosis by Arthur B Pardee Book 2 editions published in in Undetermined and English and held by 2 WorldCat member libraries worldwide. The effect of the feedback inhibitor, CTP, on subunit interactions in aspartate transcarbamylase by John Gerhart Book 1 edition published in in English and held by 1 WorldCat member library worldwide.

Part I. The degree of association of some simple antigens. Part II. The reactions of synthetic antigens with specifically purified antibodies. Part III. The size and shape of molecules of "A" substance from hog gastric mucin by Arthur B Pardee 1 edition published in in Undetermined and held by 1 WorldCat member library worldwide No Abstract Submitted. Audience Level. Related Identities.

Associated Subjects.


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