Establishment and Molecular Cytogenetic Characterization of a Cell Culture Model of Head and Neck Squamous Cell Carcinoma (HNSCC)
Abstract
:1. Introduction
2. Results and Discussion
2.1. Cytogenetic Characterization of CAL 33 Cells and Derived Cell Clones After Gene Transfection
2.1.1. Structural Rearrangements Detected by Spectral Karyotyping (SKY)
Cell Line | Chromosomal Changes Detected by SKY | Copy Number Changes Detected by array CGH | Validation by FISH | |
---|---|---|---|---|
DNA Gains | DNA Losses | |||
CAL 33 | Px + 2: 49,Y,Y,der(X)t(X;16)(p22;?), der(3)t(3;20)(p25;?), i(7)(p10),i(8)(q10), der(18)t(18;9)(p13;?)t(18;9)(q21;?),+7,+20 | Px + 2: 3q, 7p, 8q, 9p24.3-22.2, 9p13.2-p11.1, 9q, 16p13.3-11.2, 20 | Px + 2: 3p, 4q34.3-q35.1, 8p, 12q24.3, 16q22.1, 18q12.2-23 | der(3)t(3p;20q); i(3q); i(7p); i(8q); der(X)t(Xp;16p); -4q35.1; -12q24.31; |
Px + 7: 49,YY,der(X)t(X;16)(p22;?), der(3)t(3;20)(p25;?),i(7)(p10),i(8)(q10), der(18)t(18;9)(p13;?)t(18;9)(q21;?),+7,+20 | Px + 13: 1p36.3-p34.1, 1p13.3-q23.3, 1q25.2-q25.3, 1q32.1, 3q, 7p, 7q11.2, 7q22.1, 8q, 9p24.3-p22.2, 9p13.3-p11.1, 9q, 11p15.5-p15.2, 11p11.2, 11q12.1-q13.5, 11q23.3-q24.2, 12q13.1-q14.1, 16p13.3-p11.2, 17p, 17q11.2-21.3, 17q22-25.3, 19p13.3-13.1, 19q13.1-q13.4, 20, 22q | Px + 13: 2p16.3-p16.2, 3p, 4q34.3-q35.1, 8p, 12q24.3, 16q21, 16q22.1,17q21.3-q22, 18q12.2-q22, 18q22-q23 | - | |
Px + 18: 49,YY,der(X)t(X;16)(p22;?), der(3)t(3;20)(p25;?),i(7)(p10),i(8)(q10), der(18)t(18;9)(p13;?)t(18;9)(q21;?),+7,+20 | Px + 24: 1p36.3-p34.1, 1p13.3-q23.1, 1q32.1-32.2, 3q, 7p, 8q12.1-q13.1, 9p23-p22.2, 9p12-p11.1, 9q, 12q13.1-q14.1, 16p13.3-p11.2, 19p13.3-13.1, 19q13.1-q13.4, 20, 22q | Px + 24: 3p, 8p, 12q24.3, 17q21.3-q22, 18q11.2-q21.3, 18q22.1-q23 | - | |
CAL 33/ FL2-14 | 85,YYY,2x der(X)t(X;16)(p22;?), +2x der(3)t(3;20)(p25;?),+2x i(7)(p10),+i(8)(q10), +2x der(18)t(18;9)(p13;?)t(18;9)(q21;?),+1,+1,+2,+4, +5,+5,+6,+7,+7,+9,+9,+10,+11,+11,+12,+13,+14,+14,+15,+16,+17,+19,+19,+20,+20,+20,+21,+21,+22,+22 | 3q, 7p, 8q, 9p24.3-22.2, 9p13-p11.1, 9q13-q22, 9q31, 9q34, 14q, 16p13.3-p11.2, 17p13-p12, 20 | 2, 3p, 4q34.3-q35.1, 8p, 12q24.3, 15q, 16q22, 17q21-q25, 18q12-q22, 18q22-q23 | - |
2.1.2. Copy Number Alterations Detected by Array CGH
2.1.3. Fine-Mapping by FISH and Complementary Data Analysis
BAC clone | Chromosome | Localization (bp)* | FISH Labeling |
---|---|---|---|
RP11-238A9 | 3p26.1 | 4516514-4676956 | Digoxigenin |
RP11-424L2 | 3p22.1 | 41012714-41179560 | Biotin |
RP11-220O14 | 3p12.3 | 77503140-77655193 | Digoxigenin |
RP11-114I8 | 3q12.2 | 99997014-100148683 | Digoxigenin |
RP11-30J14 | 3q25.1 | 151663244-151830918 | Biotin |
RP11-392H18 | 3q28 | 190013809-190173313 | Digoxigenin |
RP11-228f3 | 4q35.1 | 185636344-185802231 | Digoxigenin |
RP11-20N02 | 7p22.3 | 758897-908887 | Biotin |
RP11-664B05 | 7q36.3 | 158712708-158902209 | Digoxigenin |
RP11-627A06 | 8q21.3 | 90721901-90763832 | Biotin |
RP11-509J21 | 9p24.2 | 3544199-3705631 | Digoxigenin |
RP11-33K8 | 9p21.3 | 24101721-24252438 | Biotin |
RP11-614P24 | 9p13.2 | 37044198-37212970 | Digoxigenin |
RP11-265B8 | 9q21.11 | 71568181-71737561 | Digoxigenin |
RP11-23B15 | 9q22.33 | 100545008-100703779 | Biotin |
RP11-269P11 | 9q33.3 | 128193295-128367188 | Digoxigenin |
RP11-158L12 | 12q24.31 | 125491151-125667388 | Digoxigenin |
RP11-22E02 | 16p13.13 | 12342021-12505232 | Biotin |
RP11-466E19 | 16q23.1 | 77319681-77480278 | Digoxigenin |
RP11-1076F02 | 18q11.2 | 19542825-19733692 | Digoxigenin |
RP11-640A09 | 20p13 | 113369-267637 | Biotin |
RP11-631L08 | 20q13.33 | 62410553-62589484 | Digoxigenin |
2.2. Establishment of a CAL 33 Cell Culture Model for Functional Studies of Radiosensitivity
3. Experimental Section
3.1. Cell Culture
3.2. Array CGH
3.3. Metaphase Preparation
3.4. Spektral Karyotyping (SKY)
3.5. FISH
3.6. Dicentric Chromosomes
3.7. Cell Survival Curves
3.8. Transfection of CAL 33
4. Conclusions
Acknowledgements
References and Notes
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Bauer, V.L.; Hieber, L.; Schaeffner, Q.; Weber, J.; Braselmann, H.; Huber, R.; Walch, A.; Zitzelsberger, H. Establishment and Molecular Cytogenetic Characterization of a Cell Culture Model of Head and Neck Squamous Cell Carcinoma (HNSCC). Genes 2010, 1, 388-412. https://doi.org/10.3390/genes1030388
Bauer VL, Hieber L, Schaeffner Q, Weber J, Braselmann H, Huber R, Walch A, Zitzelsberger H. Establishment and Molecular Cytogenetic Characterization of a Cell Culture Model of Head and Neck Squamous Cell Carcinoma (HNSCC). Genes. 2010; 1(3):388-412. https://doi.org/10.3390/genes1030388
Chicago/Turabian StyleBauer, Verena L., Ludwig Hieber, Quirin Schaeffner, Johannes Weber, Herbert Braselmann, Reinhard Huber, Axel Walch, and Horst Zitzelsberger. 2010. "Establishment and Molecular Cytogenetic Characterization of a Cell Culture Model of Head and Neck Squamous Cell Carcinoma (HNSCC)" Genes 1, no. 3: 388-412. https://doi.org/10.3390/genes1030388