SECTION DISORDERS OF EPIDERMAL AND 8 DERMAL-EPIDERMAL COHESION AND
Comparison of epidermal growth factor receptor expression in lung cancer by immunohistochemical...
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263 Basic biology Ha-ras-1 alleles in Norwegian lung cancer patients Ryberg D, Terre. T, Ovreto S et al. Depanmenr ofToxicology. Nafiond Instirute of Occvporioaal Health. P.O. Box 8149, N-0033 Oslo I. Hum Genet 1990$6:40-l. We have examined DNA restrictionfragment length polymorphisms (RFLP) of the Ha-ras- I gene in DNA from 118 lung cancer patients and 123 ttnaffectedcontmls. When DNA samples weredigested with MspU HpalI restriction endonucleases, Southern blot analysis demonstrated 4 common, 4 intemtediate and 7 different rare alleles in the combined population after hybridization to the pGDal probes. Six of the rare alleles were unique for the lung cancer group and 1 rare allele for the controlgroup. The frequencyof rare alleles in lung cancer patients(IO/ 236) was significantly different (P < 0.01) from the control group (I/ 246).ThelungcancergroupaJso hadasignificantly lower frequencyof the. common 2.57 kb fragment than the controls(P < 0.02). The results thusindicatethat Ha-ras genotyping may be of value in lung cancer risk assessment. Comparison ofepidermal growth factor receptor expressian in lung cancer by immunobistocbemicaI staining of botb froaa and for- malin fiied specimens Daazi H, Has&on PS, Thatcher N, Wilkes S, Barnes D. G&lick WJ. Department of His~oporhology.Wythenshawe Hospital, Manchester M23 9LT. Anal Cell Path01 1991;3:69-75. Two monoclonal antibodiesused to investigatethe expression of the epidermal growth factor receptor (GF-R) in 45 lung cancers were comparedTheR1 antibndyrecognisestheextracellulardomainportion of the receptor and the F4 is directed against the cytoplasmicportion. The reactivities of the two antibodies have been compared in fresh frozen tumour specimens. In addition dte staining activity of the F4 antibody(the first to EGF-R which can be used in archival material) was compared using frozen and paraffin sections from die same series of turnouts. Comparisonof the numbersof cells stainingwith the RI and F4 antibody showed only slight discrepancy when fresh material was examined. The discrepancy that existed could be explained by the the F4”antib;dy on paraffin emb&led and fresh non-small cell lung cancer material. We conclude that the expression FGF-R can be detected reliably by the F4 antibody in both the fresh frozen and formalin fixed, paraffin embedded material and could be useful in assessingtheclinical impnrtanceofEGF-R in archival tumourmaterial. Cytogenetic abnormalities and overexpression of receptors for growth factors in normal bronchial epithelium and tumor samples of lung cancer patients Sozxi G. Miozzo hi, Tagliabue E, Caldemne C, Lombardi L, Pilotti S et al. Division of Experimenral Oncology, Isrirrrro Nazionale Tumori. Via G. Venetian 1.20133 Milan. Cancer Res 1991;51:4tXl-4. A cytogenetic analysis was performed on direct preparationsand shortterm cell cultures of lung tumor and normal bronchial epidtelium of 19 patientscarrying either a first or a second primary lung cancer. In 9 tumors (6 squamous cell carcinomas, 1 adenocarcinoma. 1 mucoep idermoidcarcinoma, and I small cell lungcarcinoma) successfully ana- lyxed, pseudodiploid and hyperdiploidkaryotypes were observedwith a heterogeneous pattern of chromosome abnormalities but with a consistent involvement (5 cases) of the short or the long arm of chromosome 3. The. normal bronchial epither rl cells had a normal karyotype in 1 I patients, whereas in 6 patients clonal and nonclonal chmmosomal abnormalities were observed. Involvement of chromo- some7 was present in 4 cases.In addition, overexpression of the growth factor receptors, epidermalgrowth factor receptorand HER-Uneu. was found in 9 of 18 tumors and in 6 of I3 bronchial epithelium samples. These findingssuggest that early genetic lesions could be presentin the normal bronchial epithclial cells that are the target of further complex and multiple geneticchangcsoccurringduringthepathogenesisoflung cancer. Inactivation of the retinob&stoma gene in a human lung carcinoma cell line detected by single-strand conformation polymorphism analysis of the polymcrase chain reaction product of cDNA Murakami Y. Katahira M, Makino R, Hayashi K. Himhashi S, Sekiya T. OncogcneDivision, NafionaI Cancer CenlerResearchlnstitnuu. 1 -I, Tsukiji, S-chome. Chuo-ku. Tokyo 104. Gncogene 1991:6:3742. Combined use of a simple, sensitive method of DNA analysis of nucleotide substitutions, namely, single-strand conformation polymor- phismanalysisofpolymerasechainreactionproducts(PCR-SSCP),attd thereversetranscriptasereaction(RT)isaneffectivemedtodformRNA analysis.We used this RT-PCR-SSCP method 10 detect abnormal reti- noblastoma(RB) gene transcriptsin human tumor cell lines. Results showedthe presence of two typesof RB gene transcripts in a giant cell lung carcinoma cell line Lu65: a minor mRNA species with a base substitution that created a stop codon in the nucleotide sequence corresponding to exon 2 of the gene, and a major species of mRNA without the nucleotidesequence corresponding to that of exon 2. PCR- SSCP analysis of the genomic DNA also revealed dtat Lu65 cells contained the mutated RB allele, but not the normalallele. These results suggestedthat in Lu65 cells, both RB alleles were inactivated. The transcript without the exon 2 sequence, probably due to ahemative splicing,was also found in all the other humancells examined, as a very minor species. Differential expressioa of nuclear envelope lamins A and C in human lung cancer cell lines Kaufmann SH. Mabry M, Jasti R, Shaper JH. Oncology Cenrer, Johns Hopkins HospiraL 600 N. Wo/fe Slreer, Ballimore, MD 2120s. Cancer Res 1991;51:581-6. The lamins, an intranuclearclass of intermediate lilamerit proteins, are major structural pmteins of the nuclear envelope. In the present study,the threeabundant mammalian lamins (lamins A, B, and C) were observed tn he present in roughly equivalent amounts in the Cam-l, Cal”-3. Hl57. and SK-MES-I non-small cell lung caucer lines. In dte small cell lung cancer lines OH-l, OH-3, NCI-H82, NCI-H209, and NCI-H249. levels of lamin B were similar D those observedin the non- small cell lines, but the levels of lamins A and C were diminished by -80%. The relalionship belween lung cancer phenotype and lamin expression was explored further in the NCI-H249 small cell line. Intm- duction of the v-ras(H) oncogene into this line gives rise to a cell line (NCI-H249ras(H)) with many features of large cell carcinoma of the lung (Falco, J. P., Baylin. S. B., Lupu. R., etal. 1. Clin. Invest., 85: 1740- 1745.1990). Concomitantwith the v-ras(H)-inducedchange in pheno- type, a >lO-fold increase in the amounts of lamins A and C was observed. Levels of the cytoplasmic intermediate filament protein vimentin also increased. In contrast, levels of a variety of nonlamin nuclear polypeptides including topoisomerase I, topoisomerase II. poly(ADP-ribose) polymerase, and the nucleolar protein B23/nucleo phosmin did not change. Comparison of polyadenylated RNA fmm NCI-H249 and NCI-H249ras(H)cells on Northern blots revealed simi- lar levels of the mRNA for lamin B but higher levels of the mRNAs for lamins A and C in dte v-rasHexpressing cell line. These observations provideevidence for differences in nuclearenvelopestructure in histol- ogically different neoplastic cells derived from the same epidrelial cell system and suggest that differences in lamina structure result from phenotype-specilic differences in lamm gene expression. “‘Rc radioimmunotherapy of small cell lung carcinoma xenografts in nude mice Beaumier PL. Venkatesan P, Vanderheyden J-L. Burgua WD, Kunz LL, Fritxberg AR et al. NeoRx Corporarion. 410 W. Harrison, Seattle, WA 98119. Cancer Res 1991;51:676-81. A i=Re-labeled monoclonal antibody (MAb), NR-LU-IO. was used for the radioimmunotherapy of a subcutaneous human small cell lung carcinoma xenograft, SHT-I, in nude mice. Biodistributionwith spe- cific and iaelevanl labeled MAb demonstratedpeak tumor uptake of 8% and 3% of the injcctcd dose/g at 2 days, respectively. Dosimetry analysis predictedtumor:whole-body radiation-absorhul dose ratiosof 2.43:l for NR-LU-IO and 0.621 for irrelevant MAb. Single-dose toxicity screeningestimated a 50% lethal dose within 30 days of 600
Transcript of Comparison of epidermal growth factor receptor expression in lung cancer by immunohistochemical...
263
Basic biology
Ha-ras-1 alleles in Norwegian lung cancer patients Ryberg D, Terre. T, Ovreto S et al. Depanmenr ofToxicology. Nafiond Instirute of Occvporioaal Health. P.O. Box 8149, N-0033 Oslo I. Hum Genet 1990$6:40-l.
We have examined DNA restriction fragment length polymorphisms (RFLP) of the Ha-ras- I gene in DNA from 118 lung cancer patients and 123 ttnaffectedcontmls. When DNA samples weredigested with MspU HpalI restriction endonucleases, Southern blot analysis demonstrated 4 common, 4 intemtediate and 7 different rare alleles in the combined population after hybridization to the pGDal probes. Six of the rare alleles were unique for the lung cancer group and 1 rare allele for the control group. The frequency of rare alleles in lung cancer patients (IO/ 236) was significantly different (P < 0.01) from the control group (I/ 246).ThelungcancergroupaJso hadasignificantly lower frequency of the. common 2.57 kb fragment than the controls (P < 0.02). The results thus indicate that Ha-ras genotyping may be of value in lung cancer risk assessment.
Comparison ofepidermal growth factor receptor expressian in lung cancer by immunobistocbemicaI staining of botb froaa and for- malin fiied specimens Daazi H, Has&on PS, Thatcher N, Wilkes S, Barnes D. G&lick WJ. Department of His~oporhology. Wythenshawe Hospital, Manchester M23 9LT. Anal Cell Path01 1991;3:69-75.
Two monoclonal antibodies used to investigate the expression of the epidermal growth factor receptor (GF-R) in 45 lung cancers were comparedTheR1 antibndyrecognisestheextracellulardomainportion of the receptor and the F4 is directed against the cytoplasmic portion. The reactivities of the two antibodies have been compared in fresh frozen tumour specimens. In addition dte staining activity of the F4 antibody (the first to EGF-R which can be used in archival material) was compared using frozen and paraffin sections from die same series of turnouts. Comparison of the numbers of cells staining with the RI and F4 antibody showed only slight discrepancy when fresh material was examined. The discrepancy that existed could be explained by the
the F4”antib;dy on paraffin emb&led and fresh non-small cell lung cancer material. We conclude that the expression FGF-R can be detected reliably by the F4 antibody in both the fresh frozen and formalin fixed, paraffin embedded material and could be useful in assessingtheclinical impnrtanceofEGF-R in archival tumourmaterial.
Cytogenetic abnormalities and overexpression of receptors for growth factors in normal bronchial epithelium and tumor samples of lung cancer patients Sozxi G. Miozzo hi, Tagliabue E, Caldemne C, Lombardi L, Pilotti S et al. Division of Experimenral Oncology, Isrirrrro Nazionale Tumori. Via G. Venetian 1.20133 Milan. Cancer Res 1991;51:4tXl-4.
A cytogenetic analysis was performed on direct preparations and shortterm cell cultures of lung tumor and normal bronchial epidtelium of 19 patients carrying either a first or a second primary lung cancer. In 9 tumors (6 squamous cell carcinomas, 1 adenocarcinoma. 1 mucoep idermoid carcinoma, and I small cell lung carcinoma) successfully ana- lyxed, pseudodiploid and hyperdiploid karyotypes were observed with a heterogeneous pattern of chromosome abnormalities but with a consistent involvement (5 cases) of the short or the long arm of chromosome 3. The. normal bronchial epither rl cells had a normal karyotype in 1 I patients, whereas in 6 patients clonal and nonclonal chmmosomal abnormalities were observed. Involvement of chromo- some 7 was present in 4 cases. In addition, overexpression of the growth factor receptors, epidermal growth factor receptor and HER-Uneu. was found in 9 of 18 tumors and in 6 of I3 bronchial epithelium samples. These findings suggest that early genetic lesions could be present in the normal bronchial epithclial cells that are the target of further complex and multiple genetic changcsoccurringduring thepathogenesisof lung cancer.
Inactivation of the retinob&stoma gene in a human lung carcinoma cell line detected by single-strand conformation polymorphism analysis of the polymcrase chain reaction product of cDNA Murakami Y. Katahira M, Makino R, Hayashi K. Himhashi S, Sekiya T. OncogcneDivision, NafionaI Cancer CenlerResearchlnstitnuu. 1 -I, Tsukiji, S-chome. Chuo-ku. Tokyo 104. Gncogene 1991:6:3742.
Combined use of a simple, sensitive method of DNA analysis of nucleotide substitutions, namely, single-strand conformation polymor- phismanalysisofpolymerasechainreactionproducts(PCR-SSCP),attd thereversetranscriptasereaction(RT)isaneffectivemedtodformRNA analysis. We used this RT-PCR-SSCP method 10 detect abnormal reti- noblastoma (RB) gene transcripts in human tumor cell lines. Results showed the presence of two types of RB gene transcripts in a giant cell lung carcinoma cell line Lu65: a minor mRNA species with a base substitution that created a stop codon in the nucleotide sequence corresponding to exon 2 of the gene, and a major species of mRNA without the nucleotide sequence corresponding to that of exon 2. PCR- SSCP analysis of the genomic DNA also revealed dtat Lu65 cells contained the mutated RB allele, but not the normal allele. These results suggested that in Lu65 cells, both RB alleles were inactivated. The transcript without the exon 2 sequence, probably due to ahemative splicing, was also found in all the other human cells examined, as a very minor species.
Differential expressioa of nuclear envelope lamins A and C in human lung cancer cell lines Kaufmann SH. Mabry M, Jasti R, Shaper JH. Oncology Cenrer, Johns Hopkins HospiraL 600 N. Wo/fe Slreer, Ballimore, MD 2120s. Cancer Res 1991;51:581-6.
The lamins, an intranuclear class of intermediate lilamerit proteins, are major structural pmteins of the nuclear envelope. In the present study, the three abundant mammalian lamins (lamins A, B, and C) were observed tn he present in roughly equivalent amounts in the Cam-l, Cal”-3. Hl57. and SK-MES-I non-small cell lung caucer lines. In dte small cell lung cancer lines OH-l, OH-3, NCI-H82, NCI-H209, and NCI-H249. levels of lamin B were similar D those observed in the non- small cell lines, but the levels of lamins A and C were diminished by -80%. The relalionship belween lung cancer phenotype and lamin expression was explored further in the NCI-H249 small cell line. Intm- duction of the v-ras(H) oncogene into this line gives rise to a cell line (NCI-H249ras(H)) with many features of large cell carcinoma of the lung (Falco, J. P., Baylin. S. B., Lupu. R., etal. 1. Clin. Invest., 85: 1740- 1745.1990). Concomitant with the v-ras(H)-inducedchange in pheno- type, a >lO-fold increase in the amounts of lamins A and C was observed. Levels of the cytoplasmic intermediate filament protein vimentin also increased. In contrast, levels of a variety of nonlamin nuclear polypeptides including topoisomerase I, topoisomerase II. poly(ADP-ribose) polymerase, and the nucleolar protein B23/nucleo phosmin did not change. Comparison of polyadenylated RNA fmm NCI-H249 and NCI-H249ras(H)cells on Northern blots revealed simi- lar levels of the mRNA for lamin B but higher levels of the mRNAs for lamins A and C in dte v-rasHexpressing cell line. These observations provideevidence for differences in nuclearenvelope structure in histol- ogically different neoplastic cells derived from the same epidrelial cell system and suggest that differences in lamina structure result from phenotype-specilic differences in lamm gene expression.
“‘Rc radioimmunotherapy of small cell lung carcinoma xenografts in nude mice Beaumier PL. Venkatesan P, Vanderheyden J-L. Burgua WD, Kunz LL, Fritxberg AR et al. NeoRx Corporarion. 410 W. Harrison, Seattle, WA 98119. Cancer Res 1991;51:676-81.
A i=Re-labeled monoclonal antibody (MAb), NR-LU-IO. was used for the radioimmunotherapy of a subcutaneous human small cell lung carcinoma xenograft, SHT-I, in nude mice. Biodistribution with spe- cific and iaelevanl labeled MAb demonstrated peak tumor uptake of 8% and 3% of the injcctcd dose/g at 2 days, respectively. Dosimetry analysis predicted tumor:whole-body radiation-absorhul dose ratios of 2.43:l for NR-LU-IO and 0.621 for irrelevant MAb. Single-dose toxicity screening estimated a 50% lethal dose within 30 days of 600
