Effect of n-butyrate on cell cycle progression and in situ chromatin structure of L1210 cells

Darzynkiewicz, Zbigniew; Traganos, Frank; Xue, Sun; Melamed, Myron R.

doi:10.1016/0014-4827(81)90006-9

Cited by 75 publications

(36 citation statements)

References 25 publications

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“…However, the mechanisms by which butyrate and other SCFA regulate cell proliferation/differentiation and apoptosis are still unclear. In this sense, it has been described that butyrate is able to block cell proliferation, mainly in the G1 phase of the cell cycle (Darzynkiewicz et al 1981). This effect could be mediated by inhibition of the histone deacetylase activity (Whitlock et al 1980), an increased cyclin D and p21 Waf1 expression (Siavoshian et al 1997a, b) or a decreased expression of the proto-oncogenes c-src and c-myc (Foss et al 1989;Souleimani and Asselin 1993).…”

Section: Discussionmentioning

confidence: 99%

The effects of short-chain fatty acids on colon epithelial proliferation and survival depend on the cellular phenotype

Comalada

Bailón

Haro

et al. 2006

J Cancer Res Clin Oncol

180

125

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Section: Discussionmentioning

confidence: 99%

The effects of short-chain fatty acids on colon epithelial proliferation and survival depend on the cellular phenotype

Comalada

Bailón

Haro

et al. 2006

J Cancer Res Clin Oncol

180

125

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“…Monitoring of the enzymatic reaction requires the staining of DNA with fluorochromes, the extent of stainability after the nuclease reaction being the parameter to be determined. Interestingly, during the initial steps of the reaction with DNAse I, a n increase rather than a decrease in ethidium bromide fluorescence intensity was observed (8,271. It was inferred that, upon initial nicking, the fluorochrome could intercalate to a greater extent as a consequence of the unwinding of chromatin fibers and/or the release of constraints against intercalation.…”

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confidence: 99%

Nuclease‐induced DNA structural changes assessed by flow cytometry with the intercalating dye propidium iodide

1991

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A flow cytometric analysis of DNA structural changes induced by cleavage with nucleases was performed on isolated HeLa nuclei by assessing changes in stainability with the DNA-specific fluorochrome propidium iodide (PI). After mild digestion with DNAse I, micrococcal nuclease, or with the single-strandspecific S1 and Neurospora crassa nucleases, fluorescence intensity of nuclei stained with PI increased by about 15-30% above the value of undigested control samples. No significant modifications were observed with the restriction enymzes Eco RI, Alu I, and Not I. The DNAse I-induced increase in fluorescence intensity was also observed with the non-intercalating dye Hoechst 33258, but not with mithramycin. Nucleaseinduced fluorescence intensity changes as determined with PI were found to be dependent on the dye concentration. A constant increase (about 20%) was measured at dye/DNA-P ratios > 0.11. Below this value (2 pg/ml PI), the fluorescence intensity of digested samples was 1530% lower than that of undigested controls. This behaviour towards intercalating dyes is similar to that of the relaxed (nicked) vs. the supercoiled (intact) form of circular DNA. These results suggest that conformation-but not sequence-specific nucleases induce a relaxation of DNA supercoils.

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“…since it is a well-known differentiation inducer (19,221. It has been reported to induce G, blockade (14), reduction of clonogenic potential and phase S decrease; these effects were considered by Wallen et al (31) as criteria of a quiescent state. We also observed a G, accumulation; when DNA content was considered in only Ki-67-positive cells, G, accumulation was only visible for the highest BUT concentration.…”

Section: Discussionmentioning

confidence: 99%

“…Different attempts were carried out to stain simultaneously cells with Ki-67 MoAb and propidium iodide (3,12,24 Ki-67 expression by differentiating agents and DNA distribution in both Ki-67-positive and -negative cells. In the experiments reported here, butyric acid (BUT) was first tested, since this compound is well known to induce differentiation and quiescence in several cell strains (14,19,22), and then the effects of adriamycin (ADM), aclacinomycin A (ACM), and fagaronine (Fine), which are known to induce differentiation of K562 cells, were examined (7,101.…”

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Alterations of growth fraction and DNA content in K562 cells by differentiating agents

et al. 1990

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The growth fraction, estimated by the monoclonal antibody Ki-67 labeling, and DNA content, assessed by ethidium bromide staining, were determined simultaneously in K562 leukemic cells by flow cytometry. A multiparametric analysis enabled the fraction of the cell population with GI, S, and G, + M contents in Ki-67-positive and Ki-67-negative cells to be evaluated.Butyric acid (BUT) was used as positive control. The fraction of Ki-positive cells decreased with the BUT concentration, while the proportion of cells with G, DNA content increased only in the Ki-negative cells.Adriamycin, aclacinomycin A, and fagaronine induced differentiation, as assessed by benzidine staining and glycophorin A expression. These drugs decreased the fraction of Ki-positive cells by more than 50% for both anthracyclines and by 25% for fagaronine. Following treatment, Ki-negative cells displayed a G,, but also a G, and a S DNA content in different proportions, indicating that induction of quiescent cells by differentiating agents is not a uniform process and is worthy of interest.Key terms: FCM, proliferation-associated antibody Ki-67, glycophorin A, erythroid differentiationIn cancer therapy, it is of interest to evaluate the tumour growth fraction (GF). It reflects the rate of tumour growth which is a factor of prognosis (8,16,29) and the fraction of cells sensitive to anticancer agents. Non-cycling cells have been described with a GI but also with a G2 and even an S DNA content, both in vitro (4,13,15) and in vivo (11); "recruitment" and "synchronizing" protocols will be better scheduled if the DNA content distributions of the proliferating and quiescent cells are known. A recent strategy in cancer chemotherapy has been to induce cell maturation and inhibit cell growth (21,23) without cell killing. In this case, it will be interesting to know precisely the cell cycle parameters, i.e., the GF and the DNA distributions of both cycling and non-cycling cells. Classical methods to determine GF by using radioactive DNA precursors (11,311, the bromodeoxyuridine (BrdU) antibody technique (3,27), or dye staining of both DNA and RNA (2,4,13) are difficult and time consuming. Recently, methods using monoclonal antibodies (MoAb) have been described. They are directed against human alpha DNA polymerase (11, a heterochromatin-associated antigen (MoAb 244-7) ( 5 ) , a proliferating cell nuclear matrix antigen (32), a nuclear antigen (M 150,000) (9), nuclear antigen p105 (161, the transferrin receptor (25), the ribonucleotide reductase M1 subunit (20), and especially a nuclear-matrix-associated proliferation-related antigen (MoAb Ki-67) (18,30). Different attempts were carried out to stain simultaneously cells with Ki-67 MoAb and propidium iodide (3,12,24). We used simultaneously Ki-67 and ethidium bromide (EB) and then separately measured the DNA content of cells according to their reactivity with Ki-67 MoAb.

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Effect of n-butyrate on cell cycle progression and in situ chromatin structure of L1210 cells

Cited by 75 publications

References 25 publications

The effects of short-chain fatty acids on colon epithelial proliferation and survival depend on the cellular phenotype

The effects of short-chain fatty acids on colon epithelial proliferation and survival depend on the cellular phenotype

Nuclease‐induced DNA structural changes assessed by flow cytometry with the intercalating dye propidium iodide

Alterations of growth fraction and DNA content in K562 cells by differentiating agents

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