Published in: J. Natl. Cancer Inst. vol. 59, no. 3, pp. 839-844 (September, 1977):

"Acridine Orange-DNA Complex in Actinic Keratosis"1, 2

Masanobu Kumakiri , M.D. 3, and Ken Hashimoto, M.D. 3

1 Received January 28, 1977; accepted April 6, 1977.
2 Supported by a medical investigatorship and designated component funds (projects 3499-01 and 3499-02) of the Veterans Administration.
3 Veterans Administration Hospital, Memphis, Tenn. 38104; and Division of Dermatology, Department of Medicine, The University of Tennessee Center for the Health Sciences, Memphis, Tenn 38163.

Address reprint requests to: Dr. Hashimoto, Research Service (151), Veterans Administration Hospital, 1030 Jefferson Avenue, Memphis, Tenn. 38104.


Abstract:

Electron microscopy processing and staining of nuclei were used to localize reaction products of acridine orange staining in actinic keratosis of human skin. Electron-dense granules about 10-100 nm in diameter were seen exclusively in the euchromatin portion of the nucleus. Almost all tumor cells had granules (mean=65; SD=26). These granules were also occasionally observed in the dermal connective tissue cells in the lesion.     However, the mean number of 10 granules seen in these cells was definitely less than that of tumor cells. Normal skin controls did not have granules except occasionally in the basal cells of the epidermis.

Introduction:

Acridine orange has been used to detect the location and number of active DNA templates in fixed cells [1] as well as in the living state [2]. This technique requires the combination of acridine orange staining and DNase digestion. If positive, it visualizes the electron-dense reaction products in the euchromatin portion of the nucleus. By this ultrastructural probe method, developed and refined by Frenster et al. [1-3], human leukemia bone marrow cells were demonstrated were demonstrated to have a high DNA template activity [1]. During cell differentiation of normal bone marrow, these positive sites decrease in number [3]. Recher et al. [4] slightly modified the Frenster technique and confirmed the presence of identical acridine orange reaction products in the cultured malignant human epithelial cell line ME-180. However, its application to well-organized tissues such as skin tumors has not been reported. In this study, we found the reaction products in the lesion of actinic keratosis in significantly greater number than in the surrounding normal skin.

...

References:

1. Frenster JH, "Electron microscopic localization of acridine orange binding to DNA within human leukemic bone marrow cells", Cancer Res. 31: 1128-1131 (1971).

2. Frenster JH, "Ultrastructural Probes of chromatin within living human lymphocytes", Nature [New Biology] 236: 175-176 (1972).

3. Nakatsu SL, Masek MA, Landrum S, and Frenster JH, "Activity of DNA templates during cell division and cell differentiation", Nature 248: 334-335 (1974).

4. Recher L, Chan H, and Sykes JA, "Comparative study of microspherules and acridine orange reaction products", J. Ultrastruct. Res. 44: 347-354 (1973).

...


Additional References:

1. Frenster JH, and Herstein PR, "Gene de-repression", New Eng. J. Med. 288: 1224-1229 (1973).

2. Frenster JH, Nakatsu SL, and Masek MA, "Ultrastructural probes of DNA templates within human bone marrow and lymph node cells", Adv. Cell Molec. Biol. 3: 1-19 (1974).

3. Frenster JH, "Ultrastructure and function of heterochromatin and euchromatin", in: Busch H, ed., "The Cell Nucleus", vol. 1, 565-580 (1974), New York, Academic Press, Inc..

4. Frenster JH, "Selective control of DNA helix openings during gene regulation", Cancer Res. 36: 3394-3398 (1976).

5. Frenster JH, Papalian MM, Masek MA, and Frenster JA, "Electron microscopic analysis of  lymph node cellular activity in Hodgkin's Disease", J. Natl. Cancer Inst. 63: 331-335 (1979).

6. Frenster JH, "Oncogenes as molecular targets within active chromatin", Clin. Cancer Res. 5: supp. 1: 3855a (1999).

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 Large cell acanthoma.
Int J Dermatol. 2003 Jan;42(1):36-9.
PMID: 12581141 [PubMed - indexed for MEDLINE]

82:  Cowden RR, Curtis SK.
 Some quantitative aspects of Acridine Orange fluorescence in unfixed,
sucrose-isolated mammalian nuclei.
Histochem J. 1976 Jan;8(1):45-9.
PMID: 1270330 [PubMed - indexed for MEDLINE]

83:  McNutt NS.
 Ultrastructural comparison of the interface between epithelium and stroma in
basal cell carcinoma and control human skin.
Lab Invest. 1976 Aug;35(2):132-42.
PMID: 134174 [PubMed - indexed for MEDLINE]

84:  Okamoto H, Endo A, Imamura S.
 Langerhans cells in the dermal infiltrate of hyperkeratosis lenticularis
perstans.
Int J Dermatol. 1986 Dec;25(10):642-5.
PMID: 3804548 [PubMed - indexed for MEDLINE]

85:  Terada T, Yamagami J, Fugimoto A, Tanaka K, Sugiura M.
 Pigmented squamous cell carcinoma of the cheek skin probably arising from solar
keratosis.
Pathol Int. 2003 Jul;53(7):468-72.
PMID: 12828613 [PubMed - indexed for MEDLINE]

86:  Morimoto Y, Saga K, Bando M, Takahashi M.
 In-vitro DNA synthesis of keratinocytes in normal human skin, psoriasis,
seborrhoeic keratosis, Bowen's disease and basal cell carcinoma.
Br J Dermatol. 1991 Jul;125(1):9-13.
PMID: 1831385 [PubMed - indexed for MEDLINE]

87:  Ranger-Moore J, Bartels PH, Bozzo P, Einspahr J, Liu Y, Saboda K, Alberts
DS.
 Karyometric analysis of actinic damage in unexposed and sun-exposed skin and in
actinic keratoses in untreated individuals.
Anal Quant Cytol Histol. 2004 Jun;26(3):155-65.
PMID: 15218692 [PubMed - indexed for MEDLINE]

88:  Briggs C, Jones M.
 SYBR Green I-induced fluorescence in cultured immune cells: a comparison with
Acridine Orange.
Acta Histochem. 2005;107(4):301-12. Epub 2005 Sep 1.
PMID: 16139877 [PubMed - indexed for MEDLINE]

89:  Lyakhovitsky A, Barzilai A, Fogel M, Trau H, Huszar M.
 Expression of e-cadherin and beta-catenin in cutaneous squamous cell carcinoma
and its precursors.
Am J Dermatopathol. 2004 Oct;26(5):372-8.
PMID: 15365368 [PubMed - indexed for MEDLINE]

90:  Ranger-Moore J, Bozzo P, Alberts D, Einspahr J, Liu Y, Thompson D, Stratton
S, Stratton MS, Bartels P.
 Karyometry of nuclei from actinic keratosis and squamous cell cancer of the
skin.
Anal Quant Cytol Histol. 2003 Dec;25(6):353-61.
PMID: 14714302 [PubMed - indexed for MEDLINE]

91:  Kiyokane K, Sakatani S, Kusakabe H, Iki M, Hirayama K, Yasuhara M, Yoshida
Y, Kono K.
 A statistical study on histopathologic findings of solar keratosis.
J Med. 1992;23(6):399-408.
PMID: 1293250 [PubMed - indexed for MEDLINE]

92:  Darzynkiewicz Z.
 Differential staining of DNA and RNA in intact cells and isolated cell nuclei
with acridine orange.
Methods Cell Biol. 1990;33:285-98. No abstract available.
PMID: 1707487 [PubMed - indexed for MEDLINE]

93:  Frank GM, Karnaukhov VN, Kolaev VA, Iashin VA.
 [Spectral characteristics of human white blood cells fluorochromed with
acridine orange]
Biofizika. 1977 Nov-Dec;22(6):1015-23. Russian.
PMID: 73388 [PubMed - indexed for MEDLINE]

94:  Kastl I, Anton-Lamprecht I.
 Bicomponent keratohyalin in normal human ridged skin.
Arch Dermatol Res. 1990;282(2):71-5.
PMID: 1693840 [PubMed - indexed for MEDLINE]

95:  Takahashi M, Horiuchi Y, Tezuka T.
 Large haematoxylin-stainable keratohyaline granules in solar keratoses:
immunohistochemical comparison using anti-Ted-H-1 antibody and antiloricrin
antibody.
Br J Dermatol. 2005 Nov;153(5):976-80.
PMID: 16225609 [PubMed - indexed for MEDLINE]

96:  Goldberg LH, Joseph AK, Tschen JA.
 Proliferative actinic keratosis.
Int J Dermatol. 1994 May;33(5):341-5.
PMID: 8039973 [PubMed - indexed for MEDLINE]

97:  Tatton NA, Kish SJ.
 In situ detection of apoptotic nuclei in the substantia nigra compacta of
1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine-treated mice using terminal
deoxynucleotidyl transferase labelling and acridine orange staining.
Neuroscience. 1997 Apr;77(4):1037-48.
PMID: 9130785 [PubMed - indexed for MEDLINE]

98:  Kosower NS, Katayose H, Yanagimachi R.
 Thiol-disulfide status and acridine orange fluorescence of mammalian sperm
nuclei.
J Androl. 1992 Jul-Aug;13(4):342-8.
PMID: 1399837 [PubMed - indexed for MEDLINE]

99:  Borisov AB, Carlson BM.
 Cell death in denervated skeletal muscle is distinct from classical apoptosis.
Anat Rec. 2000 Mar 1;258(3):305-18.
PMID: 10705351 [PubMed - indexed for MEDLINE]

100:  Bolund L, De Bault LE, Foley GE, Gahrton G, Killander D, Moloney WC,
Rigler R, Skarin A, Wahren B.
 Influence of cell density on the acridine orange binding to
deoxyribonucleoprotein complex in leucocytes from patients with infectious
mononucleosis and acute leukaemia.
Acta Haematol. 1972;48(4):227-33. No abstract available.
PMID: 4117903 [PubMed - indexed for MEDLINE]

101:  Hara H, Moriki T, Hiroi M, Yamane T.
 Effects of aphidicolin and alpha-amanitin on visualization of acridine orange
binding to DNA in rat astrocytoma C6 cells.
Pathol Res Pract. 1985 Mar;179(4-5):499-503.
PMID: 2582404 [PubMed - indexed for MEDLINE]

102:  Moriki T, Hiroi M, Yamane T, Hara H.
 Effects of actinomycin D on localization of acridine orange chromatin
interaction complex in rat astrocytoma C6 cells.
Acta Pathol Jpn. 1986 Mar;36(3):389-98.
PMID: 3716793 [PubMed - indexed for MEDLINE]

103:  Zeitz M, Lange K, Noske W, Keller K, Herken H.
 Further studies on the nature of red fluorescent structures in neuroblastoma
monolayer cells vitally stained with acridine orange.
Naunyn Schmiedebergs Arch Pharmacol. 1980 Feb;311(1):53-9.
PMID: 6154246 [PubMed - indexed for MEDLINE]

104:  Rollman O, Vahlquist A.
 Cutaneous vitamin A levels in seborrheic keratosis, actinic keratosis, and
basal cell carcinoma.
Arch Dermatol Res. 1981;270(2):193-6. No abstract available.
PMID: 6454396 [PubMed - indexed for MEDLINE]

105:  Chesnokova MP, Bondarev GN.
 [Fluorochroming of yeast cells with acridine yperite]
Mikrobiologiia. 1975 Mar-Apr;44(2):367-9. Russian.
PMID: 58366 [PubMed - indexed for MEDLINE]

106:  Zs-Nagy I.
 Experimental gerontology and quantitative cytology: a new approach to cellular
ageing.
Aktuelle Gerontol. 1977 Jan;7(1):11-4.
PMID: 65923 [PubMed - indexed for MEDLINE]

Papers of M. Kumakiri listed in PubMed: January 26, 2007:
 
 
 
 
 
 
 


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