American Journal of Medical Genetics 42:217-218 (1992)
Letter to the Editor Chromosome Abnormalities in Cell Cultures Derived From the Leukoplakia of a Female Patient With Dyskeratosis Congenita To the Editor: In their letter on chromosome abnormalities in dyskeratosis congenita (DKC), Scappaticci et al. [19891reported on the occurrence of chromosome rearrangements and of an increased rate of chromosome breakage in cultures of fibroblast-like cells derived from unspecified lesions of a DKC patient. Meanwhile, about 20 publications have reported on spontaneous andor induced chromosome aberrations in cell cultures derived from male and female patients with DKC [Inoue et al., 1973; Morrison, 1974; Schroeder and Hofbauer, 1974; Sirinavin and Trowbridge, 1975; Burgdorf et al., 1977; Gutman et al., 1978; Carter et al., 1979; Connor and Teague, 1981; Kano and Fujiwara, 1982; Womer et al., 1983;Juneja et al., 1987; Aguilar-Martinez et al., 1988; Schneider et al., 1988;Scappaticci et al., 1989;Pai et al., 1989a,b; De Bauche et al., 1990; Kehrer et al., submitted]. The various forms of this disease may therefore belong to the group of precancerous syndromes characterized by chromosome instability. Cytogenetic investigations of definite precancerous lesions occurring in DKC may contribute to our understanding of instability as a possible factor predisposing to malignant degeneration. Leukoplakias are benign hyperkeratotic lesions occurring in the oral and other kinds of mucosa of DKC patients with a high frequency [Davidson and Connor, 19881. Leukoplakias undergo malignant degeneration to squamous cell carcinoma usually after a latency period of more than a decade. We established primary cultures from biopsies of a patch of oral leukoplakia from a 23-year-old woman with severe DKC. The morphology ofthe cultured cells ranged from fibroblastoid to slightly epithelioid; we did not succeed in obtaining pure cultures of keratinocytes. The results of cytogenetic analyses performed on the various cultures are summarized in Table I. Rearrangements were present in the cultures from all 3 fragments of the leukoplakia investigated and the
frequency of breakage events causing unstable anomalies was significantly increased. The presence of apparently stable dicentrics in clonal rearrangements is notable. We have further seen the following peculiarities: decondensation of 9q and l q (twice) and of an unidentified marker; pulverization of 3q and of two unidentified chromosomes (S-phase PCC?); and partial endoreduplication 18q. Interestingly, chromosome l q was involved in 3 of the 7 abnormal cells described by Scappaticci et al. [1989], and dicentrics occurred in 2 of them. The rate of tetraploid cells determined by screening of a separate sample of 866 mitoses, pooled from all cultures, was 19%,more than a third of which represented endoreduplications. Likewise, 5 of the 8 octoploid mitoses observed were endoreduplications. The occurrence in the leukoplakia cultures of various clones with structural and numerical abnormalities is highly reminiscent of the profile of cytogenetic anomalies found in non-invasive and in malignant squamous cell carcinomas of the mucosa of the respiratory and digestive tracts [Jin et al., 1988a,b; 1990a,b; Garewal et al., 19891and of the skin [Heim et al., 1988,1989;Aledo et al., 19881 respectively.
REFERENCES
Aledo R, Aurias A, Chr6tien B, Dutrillaux B (1988):Jumping translocations of chromosome 14 in a skin squamous cell carcinoma from a xeroderma pigmentosum patient. Cancer Genet Cytogenet 33:2933 Aguilar-Martinez A, Lautre-Ecenarro MJ, Urbina-Gonzllez F, Crist6bal-Gil MC, Guerra-Rodriguez P, Garcia-Perez A (1988):Cytogenetic abnormalities in dyskeratosis congenita-Report of five cases. Clin Exp Dermatol 13:lOO-104. BurgdorfW, Kurvink K, Cervenka J (1977):Sister chromatidexchange in dyskeratosis congenita lymphocytes. J Med Genet 14256-257. Carter DM, Pan M, Gaynor A, McGuire JS, Sibrack L (1979):PsoralenDNA cross-linking photoadducts in dyskeratosis congenita: Delay in excision and promotion of sister chromatid exchange. J Invest Dermatol 73:97-101. Connor JM, Teague R H (1981): Dyskeratosis congenita. Report of a large kindred. Br J Dermatol 105:321-325. Davidson HR, Connor JM (1988):Dyskeratosis congenita. J Med Genet 25~843-846. DeBauche DM, Pai GS, Stanley WS (1990): Enhanced G, chromatid radiosensitivity in dyskeratosis congenita fibroblasts. Am J Hum Received for publication March 28, 1991. Genet 46:350-357. Address reprint requests to Dr. Bruno Dallapiccola, Department Garewal HS, Sampliner R, Liu Y, Trent JM (1989): Chromosomal of Public Health and Cell Biology, I1 Universita di Roma, Via 0. rearrangements in Barrett's esophagus. A premalignant lesion of esophageal adenocarcinoma. Cancer Genet Cytogenet 42281-296. Raimondo, 00173 Rome, Italy.
0 1992 Wiley-Liss, Inc.
Kehrer and Krone
218
TABLE I. S u m m a r y of the Cytogenetic Results Obtained With Cell Cultures Derived F'rom Three Fragments of an Oral Leukoplakia of a Woman With Dyskeratosis Congenita Cells with non-
Cultures Fragment
number
clonal
were examined
No. of
No. of
at in uitro passage No.
mitoses analysed
euploid mitoses (%)
Breakage events in unstable anomalies (csb, ctb, f, dic, r)l
90 (85.7) 87 (57.2)
1 4 (13.3%) 52 (34.2%)
1
1, 3
105
2
2, 3, 5
152
3
2, 3
rearrangements and/or numerical
anomalies 2 (1.9%) 11 (7.2%)
Clonal rearrangements a n d aneuploidies (no. of cells per clone)
1:45,XX,dic(3;12)(qter;pter) (3) 91,XXXX,dict3;3)(qter;qter) (2) 90,XXXX,dic(3;12)(qter;pter) (2)
81
52 164.2)
22 (27.2%)
6 (7.4%)
2:46,X,t(1;7)(~33;933), t(5;X)(q34;q21) 46,XX,t(2;16)(pl5;q24) 4 7 , x x , 20 9 4 , x x x x , + 20, 20
+
+
(37) (16) (14) (2)
"csb, chromosome break; ctb, chromatid break; f, fragment; dic, dicentric; r, ring chromosome
Gutman A, Fmmkin A, Adam A, Bloch-Shtacher N, Rozenszajn LA (1978): X-linked dyskeratosis congenita with pancytopenia. Arch Dermatol 114:1667-1671. Heim S, Jin Y, Mandahl N, Biorklund A, Wennerberg J , Jonsson N, Mitelman F (1988):Multiple unrelated clonal chromosome abnormalities in an in situ squamous cell carcinoma of the skin. Cancer Genet Cytogenet 36:149-153. Heim S, Mertens F, Jin Y, Mandahl N, Johansson B, Biorklun? A, Wennerberg J , Jonsson N, Mitelman F (1989):Diverse chromosome abnormalities in squamous cell carcinomas of the skin. Cancer Genet Cytogenet 39:69-76. Inoue S, Mekanik G, Mahallati M, Zuelzer WW (1973): Dyskeratosis congenita with pancytopenia. Am J Dis Child 126:389-396. Jin Y, Mandahl N, Heim S, Biorklund A, Wennerberg J , Mitelman F (1988a): Unique karyotypic abnormalities in a squamous cell carcinoma of the larynx. Cancer Genet Cytogenet 30177-179. Jin Y, Heim S, Mandahl N, Biorklund A, Wennerberg J , Mitelman F (1988b):Multiple apparently unrelated clonal chromosome abnormalities in a squamous cell carcinoma of the tongue. Cancer Genet Cytogenet 3293-100. Jin Y, Heim S, Mandahl N, Biorklund A, Wennerberg J , Mitelman F (1990a):Multiple clonal chromosome aberrations in squamous cell carcinomas of the larynx. Cancer Genet Cytogenet 44:209-216. Jin Y, Heim S, Mandahl N, Biorklund A, Wennerberg J , Mitelman F (1990b): Unrelated clonal chromosomal aberrations in carcinomas of the oral cavity. Genes Chromosomes Cancer 1:209-215. Juneja HS, Elder FFB, Gardner FH (1987):Abnormality ofplatelet size and T-lymphocyte proliferation in a n autosomal recessive form of dyskeratosis congenita. Eur J Haematol 39:306-310. Kano Y, Fujiwara Y (1982): Dyskeratosis congenita: Survival, sisterchromatid exchange, and repair following treatments with crosslinking agents. Mutat Res 103:327-332.
Kehrer H, Krone W (1991): Chromosome abnormalities in cell cultures derived from a leukoplakia of a female patient with dyskeratosis congenita. (Submitted for publication). MorrisonJGL (1974):Dyskeratosis congenita. Two extremes. SAfr Med J 48:223-225. Pai GS, Morgan S, Whetsell C (1989a): Etiologic heterogeneity in dyskeratosis congenita. Am J Med Genet 32:63-66. Pai GS, Yan Y, DeBauche DM, Stanley WS, Paul SR(1989b):Bleomycin hypersensitivity in dyskeratosis congenita fibroblasts, lymphocytes, and transformed lymphoblasts. Cytogenet Cell Genet 52:186-189. Scappaticci S , Fraccaro M, Cerimele D (1989):Chromosome abnormalities in dyskeratosis congenita. Am J Med Genet 34:609-610. Schneider A, Mayer U, Gebhart E, Harms D, Gromball J , Blockel U, Beck J D (1988):Clastogen-induced fragility may differentiate pancytopenia of congenital dyskeratosis from Fanconi anaemia. Eur J Pediatr 148:37-39. Schroeder TM, Hofbauer M (1974):Letter to the editor. Dermatologica 148:316-317. Sirinavin C, nowbridge AA (1975): Dyskeratosis congenita: Clinical features and genetic aspects. Report of a family and review of the literature. J Med Genet 12:339-354. Womer R, Clark JE, Wood P, Sabio H, Kelly TE (1983): Dyskeratosis congenita: Two examples of this multisystem disorder. Pediatrics 71:603-609.
Hildegard Kehrer Winfrid Krone Abteilung Humangenetik, Universitat Ulm, 79 Ulm, Germany
Letter to the Editor Chromosome Abnormalities in Cell Cultures Derived From the Leukoplakia of a Female Patient With Dyskeratosis Congenita To the Editor: In their letter on chromosome abnormalities in dyskeratosis congenita (DKC), Scappaticci et al. [19891reported on the occurrence of chromosome rearrangements and of an increased rate of chromosome breakage in cultures of fibroblast-like cells derived from unspecified lesions of a DKC patient. Meanwhile, about 20 publications have reported on spontaneous andor induced chromosome aberrations in cell cultures derived from male and female patients with DKC [Inoue et al., 1973; Morrison, 1974; Schroeder and Hofbauer, 1974; Sirinavin and Trowbridge, 1975; Burgdorf et al., 1977; Gutman et al., 1978; Carter et al., 1979; Connor and Teague, 1981; Kano and Fujiwara, 1982; Womer et al., 1983;Juneja et al., 1987; Aguilar-Martinez et al., 1988; Schneider et al., 1988;Scappaticci et al., 1989;Pai et al., 1989a,b; De Bauche et al., 1990; Kehrer et al., submitted]. The various forms of this disease may therefore belong to the group of precancerous syndromes characterized by chromosome instability. Cytogenetic investigations of definite precancerous lesions occurring in DKC may contribute to our understanding of instability as a possible factor predisposing to malignant degeneration. Leukoplakias are benign hyperkeratotic lesions occurring in the oral and other kinds of mucosa of DKC patients with a high frequency [Davidson and Connor, 19881. Leukoplakias undergo malignant degeneration to squamous cell carcinoma usually after a latency period of more than a decade. We established primary cultures from biopsies of a patch of oral leukoplakia from a 23-year-old woman with severe DKC. The morphology ofthe cultured cells ranged from fibroblastoid to slightly epithelioid; we did not succeed in obtaining pure cultures of keratinocytes. The results of cytogenetic analyses performed on the various cultures are summarized in Table I. Rearrangements were present in the cultures from all 3 fragments of the leukoplakia investigated and the
frequency of breakage events causing unstable anomalies was significantly increased. The presence of apparently stable dicentrics in clonal rearrangements is notable. We have further seen the following peculiarities: decondensation of 9q and l q (twice) and of an unidentified marker; pulverization of 3q and of two unidentified chromosomes (S-phase PCC?); and partial endoreduplication 18q. Interestingly, chromosome l q was involved in 3 of the 7 abnormal cells described by Scappaticci et al. [1989], and dicentrics occurred in 2 of them. The rate of tetraploid cells determined by screening of a separate sample of 866 mitoses, pooled from all cultures, was 19%,more than a third of which represented endoreduplications. Likewise, 5 of the 8 octoploid mitoses observed were endoreduplications. The occurrence in the leukoplakia cultures of various clones with structural and numerical abnormalities is highly reminiscent of the profile of cytogenetic anomalies found in non-invasive and in malignant squamous cell carcinomas of the mucosa of the respiratory and digestive tracts [Jin et al., 1988a,b; 1990a,b; Garewal et al., 19891and of the skin [Heim et al., 1988,1989;Aledo et al., 19881 respectively.
REFERENCES
Aledo R, Aurias A, Chr6tien B, Dutrillaux B (1988):Jumping translocations of chromosome 14 in a skin squamous cell carcinoma from a xeroderma pigmentosum patient. Cancer Genet Cytogenet 33:2933 Aguilar-Martinez A, Lautre-Ecenarro MJ, Urbina-Gonzllez F, Crist6bal-Gil MC, Guerra-Rodriguez P, Garcia-Perez A (1988):Cytogenetic abnormalities in dyskeratosis congenita-Report of five cases. Clin Exp Dermatol 13:lOO-104. BurgdorfW, Kurvink K, Cervenka J (1977):Sister chromatidexchange in dyskeratosis congenita lymphocytes. J Med Genet 14256-257. Carter DM, Pan M, Gaynor A, McGuire JS, Sibrack L (1979):PsoralenDNA cross-linking photoadducts in dyskeratosis congenita: Delay in excision and promotion of sister chromatid exchange. J Invest Dermatol 73:97-101. Connor JM, Teague R H (1981): Dyskeratosis congenita. Report of a large kindred. Br J Dermatol 105:321-325. Davidson HR, Connor JM (1988):Dyskeratosis congenita. J Med Genet 25~843-846. DeBauche DM, Pai GS, Stanley WS (1990): Enhanced G, chromatid radiosensitivity in dyskeratosis congenita fibroblasts. Am J Hum Received for publication March 28, 1991. Genet 46:350-357. Address reprint requests to Dr. Bruno Dallapiccola, Department Garewal HS, Sampliner R, Liu Y, Trent JM (1989): Chromosomal of Public Health and Cell Biology, I1 Universita di Roma, Via 0. rearrangements in Barrett's esophagus. A premalignant lesion of esophageal adenocarcinoma. Cancer Genet Cytogenet 42281-296. Raimondo, 00173 Rome, Italy.
0 1992 Wiley-Liss, Inc.
Kehrer and Krone
218
TABLE I. S u m m a r y of the Cytogenetic Results Obtained With Cell Cultures Derived F'rom Three Fragments of an Oral Leukoplakia of a Woman With Dyskeratosis Congenita Cells with non-
Cultures Fragment
number
clonal
were examined
No. of
No. of
at in uitro passage No.
mitoses analysed
euploid mitoses (%)
Breakage events in unstable anomalies (csb, ctb, f, dic, r)l
90 (85.7) 87 (57.2)
1 4 (13.3%) 52 (34.2%)
1
1, 3
105
2
2, 3, 5
152
3
2, 3
rearrangements and/or numerical
anomalies 2 (1.9%) 11 (7.2%)
Clonal rearrangements a n d aneuploidies (no. of cells per clone)
1:45,XX,dic(3;12)(qter;pter) (3) 91,XXXX,dict3;3)(qter;qter) (2) 90,XXXX,dic(3;12)(qter;pter) (2)
81
52 164.2)
22 (27.2%)
6 (7.4%)
2:46,X,t(1;7)(~33;933), t(5;X)(q34;q21) 46,XX,t(2;16)(pl5;q24) 4 7 , x x , 20 9 4 , x x x x , + 20, 20
+
+
(37) (16) (14) (2)
"csb, chromosome break; ctb, chromatid break; f, fragment; dic, dicentric; r, ring chromosome
Gutman A, Fmmkin A, Adam A, Bloch-Shtacher N, Rozenszajn LA (1978): X-linked dyskeratosis congenita with pancytopenia. Arch Dermatol 114:1667-1671. Heim S, Jin Y, Mandahl N, Biorklund A, Wennerberg J , Jonsson N, Mitelman F (1988):Multiple unrelated clonal chromosome abnormalities in an in situ squamous cell carcinoma of the skin. Cancer Genet Cytogenet 36:149-153. Heim S, Mertens F, Jin Y, Mandahl N, Johansson B, Biorklun? A, Wennerberg J , Jonsson N, Mitelman F (1989):Diverse chromosome abnormalities in squamous cell carcinomas of the skin. Cancer Genet Cytogenet 39:69-76. Inoue S, Mekanik G, Mahallati M, Zuelzer WW (1973): Dyskeratosis congenita with pancytopenia. Am J Dis Child 126:389-396. Jin Y, Mandahl N, Heim S, Biorklund A, Wennerberg J , Mitelman F (1988a): Unique karyotypic abnormalities in a squamous cell carcinoma of the larynx. Cancer Genet Cytogenet 30177-179. Jin Y, Heim S, Mandahl N, Biorklund A, Wennerberg J , Mitelman F (1988b):Multiple apparently unrelated clonal chromosome abnormalities in a squamous cell carcinoma of the tongue. Cancer Genet Cytogenet 3293-100. Jin Y, Heim S, Mandahl N, Biorklund A, Wennerberg J , Mitelman F (1990a):Multiple clonal chromosome aberrations in squamous cell carcinomas of the larynx. Cancer Genet Cytogenet 44:209-216. Jin Y, Heim S, Mandahl N, Biorklund A, Wennerberg J , Mitelman F (1990b): Unrelated clonal chromosomal aberrations in carcinomas of the oral cavity. Genes Chromosomes Cancer 1:209-215. Juneja HS, Elder FFB, Gardner FH (1987):Abnormality ofplatelet size and T-lymphocyte proliferation in a n autosomal recessive form of dyskeratosis congenita. Eur J Haematol 39:306-310. Kano Y, Fujiwara Y (1982): Dyskeratosis congenita: Survival, sisterchromatid exchange, and repair following treatments with crosslinking agents. Mutat Res 103:327-332.
Kehrer H, Krone W (1991): Chromosome abnormalities in cell cultures derived from a leukoplakia of a female patient with dyskeratosis congenita. (Submitted for publication). MorrisonJGL (1974):Dyskeratosis congenita. Two extremes. SAfr Med J 48:223-225. Pai GS, Morgan S, Whetsell C (1989a): Etiologic heterogeneity in dyskeratosis congenita. Am J Med Genet 32:63-66. Pai GS, Yan Y, DeBauche DM, Stanley WS, Paul SR(1989b):Bleomycin hypersensitivity in dyskeratosis congenita fibroblasts, lymphocytes, and transformed lymphoblasts. Cytogenet Cell Genet 52:186-189. Scappaticci S , Fraccaro M, Cerimele D (1989):Chromosome abnormalities in dyskeratosis congenita. Am J Med Genet 34:609-610. Schneider A, Mayer U, Gebhart E, Harms D, Gromball J , Blockel U, Beck J D (1988):Clastogen-induced fragility may differentiate pancytopenia of congenital dyskeratosis from Fanconi anaemia. Eur J Pediatr 148:37-39. Schroeder TM, Hofbauer M (1974):Letter to the editor. Dermatologica 148:316-317. Sirinavin C, nowbridge AA (1975): Dyskeratosis congenita: Clinical features and genetic aspects. Report of a family and review of the literature. J Med Genet 12:339-354. Womer R, Clark JE, Wood P, Sabio H, Kelly TE (1983): Dyskeratosis congenita: Two examples of this multisystem disorder. Pediatrics 71:603-609.
Hildegard Kehrer Winfrid Krone Abteilung Humangenetik, Universitat Ulm, 79 Ulm, Germany
