Analysis of K-ras, p53, bcl-2 and Rb expression in non-small cell lung cancer cell lines

M. Loprevite, L. Varesco, R. Favoni, G. B. Ferrara, F. Moro, L. Ottaggio, G. Fronza, P. Campomenosi, A. Abbondandolo, G. Cutrona, S. Roncella, A. Albini, M. G. Aluigi, S. Pozzi, C. Pera, R. Biticchi, V. Gismondi, F. Grossi, M. C. Pennucci, A. Ardizzoni

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Abstract

Six non-small cell lung cancer (NSCLC) cell lines (A-549, Ca-Lu-6, SK- Lu-1, Ca-Lu-1, SK-Mes-1 and LX-1) were studied to assess the presence of multiple concomitant alterations of different oncogenes (K-ras, bcl-2) and tumor suppressor genes (p53, Rb) in NSCLC. K-ras (exon 1) and p53 (exons 5- 8) gene mutations were determined via a PCR-based-DGGE (Denaturing Gradient Gel Electro-phoresis) and by sequencing approach. Different mutations were found in the 1st exon of K-ras gene in 5 of 6 cell lines examined. Five of six cell lines contained K-ras mutations at codon 12 (A-549, SK-Lu-1, LX-1) or codon 13 (SK-Mes-1, Ca-Lu-1). In addition, 5 of 6 cell lines showed p53 mutations of exon 8 (SK-Mes-1, Ca-Lu-1 cod. 280; LX-1 cod. 273) or exon 6 (Ca-Lu-6 cod. 196; SK-Lu-1 cod. 193). In 4 of these cell lines, p53 protein nuclear expression was also confirmed with DO-7 mAb immunocytochemistry. Expression of cytoplasmic bcl-2 protein, by anti-bcl-2 mAb flow cytometric analysis, was found in A-549, Ca-Lu-1, SK-Lu-1, SK-Mes-1 cell lines. In contrast, RT-PCR analysis of Rb gene could not identify any change in the cell lines examined. In conclusion, most NSCLC cell lines tested displayed concomitant multiple oncogene/tumor suppressor gene alterations.

Original languageEnglish
Pages (from-to)1203-1208
Number of pages6
JournalInternational Journal of Oncology
Volume11
Issue number6
Publication statusPublished - 1997

Fingerprint

Non-Small Cell Lung Carcinoma
Gadiformes
Cell Line
Exons
Mutation
ras Genes
Tumor Suppressor Genes
Codon
Retinoblastoma Genes
Polymerase Chain Reaction
Nuclear Proteins
Oncogenes
Gels
Immunohistochemistry
Genes
Proteins

Keywords

  • Bcl-2
  • K-ras
  • Non-small cell lung cancer
  • P53
  • Rb

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Loprevite, M., Varesco, L., Favoni, R., Ferrara, G. B., Moro, F., Ottaggio, L., ... Ardizzoni, A. (1997). Analysis of K-ras, p53, bcl-2 and Rb expression in non-small cell lung cancer cell lines. International Journal of Oncology, 11(6), 1203-1208.

Analysis of K-ras, p53, bcl-2 and Rb expression in non-small cell lung cancer cell lines. / Loprevite, M.; Varesco, L.; Favoni, R.; Ferrara, G. B.; Moro, F.; Ottaggio, L.; Fronza, G.; Campomenosi, P.; Abbondandolo, A.; Cutrona, G.; Roncella, S.; Albini, A.; Aluigi, M. G.; Pozzi, S.; Pera, C.; Biticchi, R.; Gismondi, V.; Grossi, F.; Pennucci, M. C.; Ardizzoni, A.

In: International Journal of Oncology, Vol. 11, No. 6, 1997, p. 1203-1208.

Research output: Contribution to journalArticle

Loprevite, M, Varesco, L, Favoni, R, Ferrara, GB, Moro, F, Ottaggio, L, Fronza, G, Campomenosi, P, Abbondandolo, A, Cutrona, G, Roncella, S, Albini, A, Aluigi, MG, Pozzi, S, Pera, C, Biticchi, R, Gismondi, V, Grossi, F, Pennucci, MC & Ardizzoni, A 1997, 'Analysis of K-ras, p53, bcl-2 and Rb expression in non-small cell lung cancer cell lines', International Journal of Oncology, vol. 11, no. 6, pp. 1203-1208.
Loprevite, M. ; Varesco, L. ; Favoni, R. ; Ferrara, G. B. ; Moro, F. ; Ottaggio, L. ; Fronza, G. ; Campomenosi, P. ; Abbondandolo, A. ; Cutrona, G. ; Roncella, S. ; Albini, A. ; Aluigi, M. G. ; Pozzi, S. ; Pera, C. ; Biticchi, R. ; Gismondi, V. ; Grossi, F. ; Pennucci, M. C. ; Ardizzoni, A. / Analysis of K-ras, p53, bcl-2 and Rb expression in non-small cell lung cancer cell lines. In: International Journal of Oncology. 1997 ; Vol. 11, No. 6. pp. 1203-1208.
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abstract = "Six non-small cell lung cancer (NSCLC) cell lines (A-549, Ca-Lu-6, SK- Lu-1, Ca-Lu-1, SK-Mes-1 and LX-1) were studied to assess the presence of multiple concomitant alterations of different oncogenes (K-ras, bcl-2) and tumor suppressor genes (p53, Rb) in NSCLC. K-ras (exon 1) and p53 (exons 5- 8) gene mutations were determined via a PCR-based-DGGE (Denaturing Gradient Gel Electro-phoresis) and by sequencing approach. Different mutations were found in the 1st exon of K-ras gene in 5 of 6 cell lines examined. Five of six cell lines contained K-ras mutations at codon 12 (A-549, SK-Lu-1, LX-1) or codon 13 (SK-Mes-1, Ca-Lu-1). In addition, 5 of 6 cell lines showed p53 mutations of exon 8 (SK-Mes-1, Ca-Lu-1 cod. 280; LX-1 cod. 273) or exon 6 (Ca-Lu-6 cod. 196; SK-Lu-1 cod. 193). In 4 of these cell lines, p53 protein nuclear expression was also confirmed with DO-7 mAb immunocytochemistry. Expression of cytoplasmic bcl-2 protein, by anti-bcl-2 mAb flow cytometric analysis, was found in A-549, Ca-Lu-1, SK-Lu-1, SK-Mes-1 cell lines. In contrast, RT-PCR analysis of Rb gene could not identify any change in the cell lines examined. In conclusion, most NSCLC cell lines tested displayed concomitant multiple oncogene/tumor suppressor gene alterations.",
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AU - Loprevite, M.

AU - Varesco, L.

AU - Favoni, R.

AU - Ferrara, G. B.

AU - Moro, F.

AU - Ottaggio, L.

AU - Fronza, G.

AU - Campomenosi, P.

AU - Abbondandolo, A.

AU - Cutrona, G.

AU - Roncella, S.

AU - Albini, A.

AU - Aluigi, M. G.

AU - Pozzi, S.

AU - Pera, C.

AU - Biticchi, R.

AU - Gismondi, V.

AU - Grossi, F.

AU - Pennucci, M. C.

AU - Ardizzoni, A.

PY - 1997

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N2 - Six non-small cell lung cancer (NSCLC) cell lines (A-549, Ca-Lu-6, SK- Lu-1, Ca-Lu-1, SK-Mes-1 and LX-1) were studied to assess the presence of multiple concomitant alterations of different oncogenes (K-ras, bcl-2) and tumor suppressor genes (p53, Rb) in NSCLC. K-ras (exon 1) and p53 (exons 5- 8) gene mutations were determined via a PCR-based-DGGE (Denaturing Gradient Gel Electro-phoresis) and by sequencing approach. Different mutations were found in the 1st exon of K-ras gene in 5 of 6 cell lines examined. Five of six cell lines contained K-ras mutations at codon 12 (A-549, SK-Lu-1, LX-1) or codon 13 (SK-Mes-1, Ca-Lu-1). In addition, 5 of 6 cell lines showed p53 mutations of exon 8 (SK-Mes-1, Ca-Lu-1 cod. 280; LX-1 cod. 273) or exon 6 (Ca-Lu-6 cod. 196; SK-Lu-1 cod. 193). In 4 of these cell lines, p53 protein nuclear expression was also confirmed with DO-7 mAb immunocytochemistry. Expression of cytoplasmic bcl-2 protein, by anti-bcl-2 mAb flow cytometric analysis, was found in A-549, Ca-Lu-1, SK-Lu-1, SK-Mes-1 cell lines. In contrast, RT-PCR analysis of Rb gene could not identify any change in the cell lines examined. In conclusion, most NSCLC cell lines tested displayed concomitant multiple oncogene/tumor suppressor gene alterations.

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