Mutation and overexpression of p53 gene in patients with primary resected non-small cell lung cancer...

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13 046 OVRRRXPRESSION OF P53 PROTRIN CORRELATES WITH PROGNOSIS IN NON SWALL CELL LUNG CANCER (NSCLC). S. Yoneda* ** T. Kusano**, A. Iwasaki**, K. Ando**, T. Sh&akusa**. *First Department of Pathology and **Second Department of Surgery, School of Medicine, Fukuoka University, Fukuoka, Japan. There is a possibility that alteration of the p53 gene is a poor prognostic factor. We analyz- ed immunohistochemically the relationship between p53 expression and clinicopathological features. We analyzed 181 non-small cell lung cancer (NSCLC) specimens during surgical resection. Immunohistochemical stainings with anti-p53 anti- body (DO?), using the ABC method were performed for methacarn-fixed, paraffin embedded tumor tissue. We classified reactive tumors into four groups according to the percentage of immunostai- ned tumor cells: Grade O<l%, Grade 1=1-30%, Grade 2=31-60%, Grade 3=61-100%. Of the 181 NSCLC specimens, 97 (54%) were positive with nuclear accumulation of pi3 protein; adenocarcinoma (Ad) were 44 (45%) of 97, squamous cell carcinoma (Sq) were 35 (65%) of 54, adenosquamous carcinoma were 12 (55%) of 22 and large cell carcinomas were 6 (75%) of 8. The frequency p53 expression was more significant in Sq than in Ad (PiO.03). Eight cases with poorly differentiated Sq were positive for p53 antibody. We analyzed the corr- elation between pi3 expression and prognosis. Patients with p53 positive tumors had significan- tly shorter survivals than those with p53 negati- ve tumors (P=O.O05). in proportion as grades of p53 expression increased, the survival rate sign- ificantly shortened (p=O.O16). 048 MUTATION AND OVEREXPRESSION OF ~53 GENE IN PATIENTS WITH PRIMARY RESECTED NON-SMALL CELL LUNG CANCER (NSCLC). A. Kubot , K. Nakagawal,. A. Yoshikawal, T. Hirashimal, T. Nittal,s, T. Yanal, N. Masuda’ , K. Matsuit, Y. Kusunokit, M. Takadal, M. FukuokalV*. ‘Department of Internal Medicine, Osaka Prefectural Habikino Hospital, Habikino, Osaka, and *Osaka City General Hospital, Miyakojima, Osaka, Japan. To investigate the relationship between mutation or expression of p53 gene, and clinical prognosis in patients with primary resected NSCLC, and to obtain basic data for prospective study to confirm their relationship, we determined p53 gene mutation by PCR-SSCP (Polymerase chain reaction - single strand conformation polymorphism) analysis and overexpression by LSAB (Labelled streptavidin biotin) assay. A total of 67 patients of primary resected NSCLC (male/female:40/27, pathological stage I/II/IIIA: 35/l 3/19) were examined by PCR-SSCP analysis, and 26 of 67 patients by immunohistochemical staining using anti-p53 monoclonal antibody (DAKO-~53, DO-7). p53 mutations were detected in 26 (39%) of 67 patients by PCR-SSCP analysis and overexpression of p53 were detected in 7 (27%) of 26 patients by immunohistochemical staining, indicating that overexpression of p53 protein was not synonymous with the presence of gene mutation, and that some mutation could not be detected by a simple immunohistochemical analysis. In this study, there were not any significant correlations between the presence of ~53 mutations and sex, age, histological subtype, pathological stage, body weight loss, serum level of CEA, and cigarette index. Our preliminary data suggested that ~53 gene mutation might be associated with poor prognosis of patients with stage I-II NSCLC. Ditierences in Allelotype between Squamous Cell Carcinomas and Adenocarcinomas of the Lung sato, s i Kawabuchl, 6.’ lshikavia Y Nziamura \ Nakagaha. KJ Tsuchiya. E ‘Dept Pamcl ‘Dept Blochemis!y, ‘Siige,-!, Cancer lnai ?x Hospital Loss of heterozygosity (LOH) was examined on all autosomal chromosomes I” 175 non-small cell lung cancers (41 squamous cell carcinomas, 119 adenocarcmomas, and 15 others) and the frequenctes compared behveen dlfferent tumor types to cast light on spectfic tumor suppressor gene(s). On the majority of chromosomal arms ( 27 of 35 ), LOH was observed mow irequently in squamous cell carcinomas than in adenocarclnomas, and on 9 chromosomal arns (3p, 3q. 4q, 7~. 9q, 13q, 16q, 17~. and 21q), the dtfference was stattstically signlficant’ (3p, 3q, 4q, 9q. 13q. and 17~. p<O.Ol, 7p, 16q, and Zlq: p<O.O5; y ‘ test) In squamous cell carcinomas. frequent allellc loss on chromosome 9q (67%) was observed, II? addition to prev~ousiy reported chromosomes 3p (82%) and 17~ (88%), followed by chromosomel3q (60%). On the other hand, I” adenocarclnomas, frequent LOH was observed on chromosome Bq (32%), after chromosomes 3p (40%) and 17p (51%). Further, allelic loss on chromosomes 9q and 13q tended to correlate with a history of smoking These results suggest that accumulative genetic change might be more directly associated Wh development of squamous cell carcinomas than adenocatcinomas, wtth possible wolvement of unidentified tumor suppressor gene(s) on chromosome 9q and chromosome 8q, respectively I” the two cases. The observed differences may be related to variation in the underlying mechanisms of tumorlgenesis, such as etiologicai factors, for the two hlstopathological disease entitles 049 p53 gene abnormalities in primary lung cancer and metsstatic site tumor Akihiko Gemna, Yasuvuki Taniguchi, Yuichiro Takeda. Kivoshi Takenaka, Tskashi Shiaada, Shoji Kudoh, Hisenobu Niitani. The Forth Dept. of Internal Medicine, Nippon Medical school, Tokyo, Japan In order to study the cccurrence rate of ~53 gene mutation in primary lung cancers after distant metestasis. 32 pairs of primary and aetastatlc tumors were analysed for abnormalities of ~53 gene by PCR-SSCP ( Polymerase Chain Reaction - Single Strand Conformation Polymorphism ). The tumors used in this study were obteined by autopsy. They consisted of six small cell carcinomas, 13 adenocarcinomas. nine swamcus cell carcinomas, and one large cell carcinoma. PCR-SSCP analysis shored that seven non saall cell carcinomas ( 27 X 1 and three small cell carcinomas ( 50 X ) had ~53 gene mutations. The abnormalities were shown at excn 5-9. All pairs of primary and aetastatic tuacrs were identical for ~53 gene. The survival curves of autation- positive and negative groups wars similar. These results indicate that ~53 gene autasion may cccurs before distant metastasis and be stable in general.

Transcript of Mutation and overexpression of p53 gene in patients with primary resected non-small cell lung cancer...

Page 1: Mutation and overexpression of p53 gene in patients with primary resected non-small cell lung cancer (NSCLC)

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046

OVRRRXPRESSION OF P53 PROTRIN CORRELATES WITH PROGNOSIS IN NON SWALL CELL LUNG CANCER (NSCLC). S. Yoneda* ** T. Kusano**, A. Iwasaki**, K. Ando**, T. Sh&akusa**. *First Department of Pathology and **Second Department of Surgery, School of Medicine, Fukuoka University, Fukuoka, Japan.

There is a possibility that alteration of the p53 gene is a poor prognostic factor. We analyz- ed immunohistochemically the relationship between p53 expression and clinicopathological features. We analyzed 181 non-small cell lung cancer (NSCLC) specimens during surgical resection. Immunohistochemical stainings with anti-p53 anti- body (DO?), using the ABC method were performed for methacarn-fixed, paraffin embedded tumor tissue. We classified reactive tumors into four groups according to the percentage of immunostai- ned tumor cells: Grade O<l%, Grade 1=1-30%, Grade 2=31-60%, Grade 3=61-100%. Of the 181 NSCLC specimens, 97 (54%) were positive with nuclear accumulation of pi3 protein; adenocarcinoma (Ad) were 44 (45%) of 97, squamous cell carcinoma (Sq) were 35 (65%) of 54, adenosquamous carcinoma were 12 (55%) of 22 and large cell carcinomas were 6 (75%) of 8. The frequency p53 expression was more significant in Sq than in Ad (PiO.03). Eight cases with poorly differentiated Sq were positive for p53 antibody. We analyzed the corr- elation between pi3 expression and prognosis. Patients with p53 positive tumors had significan- tly shorter survivals than those with p53 negati- ve tumors (P=O.O05). in proportion as grades of p53 expression increased, the survival rate sign- ificantly shortened (p=O.O16).

048

MUTATION AND OVEREXPRESSION OF ~53 GENE IN PATIENTS WITH PRIMARY RESECTED NON-SMALL CELL LUNG CANCER (NSCLC). A. Kubot , K. Nakagawal,. A. Yoshikawal, T. Hirashimal, T. Nittal,s, T. Yanal, N. Masuda’, K. Matsuit, Y. Kusunokit, M. Takadal, M. FukuokalV*. ‘Department of Internal Medicine, Osaka Prefectural Habikino Hospital, Habikino, Osaka, and *Osaka City General Hospital, Miyakojima, Osaka, Japan.

To investigate the relationship between mutation or expression of p53 gene, and clinical prognosis in patients with primary resected NSCLC, and to obtain basic data for prospective study to confirm their relationship, we determined p53 gene mutation by PCR-SSCP (Polymerase chain reaction - single strand conformation polymorphism) analysis and overexpression by LSAB (Labelled streptavidin biotin) assay. A total of 67 patients of primary resected NSCLC (male/female:40/27, pathological stage I/II/IIIA: 35/l 3/19) were examined by PCR-SSCP analysis, and 26 of 67 patients by immunohistochemical staining using anti-p53 monoclonal antibody (DAKO-~53, DO-7).

p53 mutations were detected in 26 (39%) of 67 patients by PCR-SSCP analysis and overexpression of p53 were detected in 7 (27%) of 26 patients by immunohistochemical staining, indicating that overexpression of p53 protein was not synonymous with the presence of gene mutation, and that some mutation could not be detected by a simple immunohistochemical analysis. In this study, there were not any significant correlations between the presence of ~53 mutations and sex, age, histological subtype, pathological stage, body weight loss, serum level of CEA, and cigarette index. Our preliminary data suggested that ~53 gene mutation might be associated with poor prognosis of patients with stage I-II NSCLC.

Ditierences in Allelotype between Squamous Cell Carcinomas

and Adenocarcinomas of the Lung

sato, s i Kawabuchl, 6.’ lshikavia Y Nziamura \ Nakagaha. KJ

Tsuchiya. E ’ ‘Dept Pamcl ‘Dept Blochemis!y, ‘Siige,-!, Cancer lnai

?x Hospital

Loss of heterozygosity (LOH) was examined on all autosomal chromosomes

I” 175 non-small cell lung cancers (41 squamous cell carcinomas, 119

adenocarcmomas, and 15 others) and the frequenctes compared behveen

dlfferent tumor types to cast light on spectfic tumor suppressor gene(s). On

the majority of chromosomal arms ( 27 of 35 ), LOH was observed mow

irequently in squamous cell carcinomas than in adenocarclnomas, and on 9

chromosomal arns (3p, 3q. 4q, 7~. 9q, 13q, 16q, 17~. and 21q), the

dtfference was stattstically signlficant’(3p, 3q, 4q, 9q. 13q. and 17~. p<O.Ol,

7p, 16q, and Zlq: p<O.O5; y ‘test) In squamous cell carcinomas. frequent

allellc loss on chromosome 9q (67%) was observed, II? addition to prev~ousiy

reported chromosomes 3p (82%) and 17~ (88%), followed by

chromosomel3q (60%). On the other hand, I” adenocarclnomas, frequent

LOH was observed on chromosome Bq (32%), after chromosomes 3p (40%)

and 17p (51%). Further, allelic loss on chromosomes 9q and 13q tended to

correlate with a history of smoking These results suggest that accumulative

genetic change might be more directly associated Wh development of

squamous cell carcinomas than adenocatcinomas, wtth possible wolvement

of unidentified tumor suppressor gene(s) on chromosome 9q and

chromosome 8q, respectively I” the two cases. The observed differences

may be related to variation in the underlying mechanisms of tumorlgenesis,

such as etiologicai factors, for the two hlstopathological disease entitles

049

p53 gene abnormalities in primary lung cancer and metsstatic

site tumor

Akihiko Gemna, Yasuvuki Taniguchi, Yuichiro Takeda. Kivoshi

Takenaka, Tskashi Shiaada, Shoji Kudoh, Hisenobu Niitani.

The Forth Dept. of Internal Medicine, Nippon Medical school,

Tokyo, Japan

In order to study the cccurrence rate of ~53 gene

mutation in primary lung cancers after distant metestasis.

32 pairs of primary and aetastatlc tumors were analysed for

abnormalities of ~53 gene by PCR-SSCP ( Polymerase Chain

Reaction - Single Strand Conformation Polymorphism ). The

tumors used in this study were obteined by autopsy. They consisted of six small cell carcinomas, 13 adenocarcinomas.

nine swamcus cell carcinomas, and one large cell carcinoma.

PCR-SSCP analysis shored that seven non saall cell

carcinomas ( 27 X 1 and three small cell carcinomas ( 50 X )

had ~53 gene mutations. The abnormalities were shown at

excn 5-9. All pairs of primary and aetastatic tuacrs were

identical for ~53 gene. The survival curves of autation-

positive and negative groups wars similar. These results

indicate that ~53 gene autasion may cccurs before distant

metastasis and be stable in general.