Hitoshi Haneda

Prognostic and therapeutic significance of the flow cytometric nuclear DNA content in non-small cell lung cancer

To evaluate prognostic and therapeutic significance, tumor DNA content was determined by flow cytometry in 310 paraffin‐embedded tissue samples obtained surgically from 130 patients with non‐small cell lung cancer. Ninety‐six (76.8%) patients had DNA aneuploid patterns that were statistically higher in adenocarcinoma than in squamous cell carcinoma. A better 5‐year survival rate was observed in Group A (DNA diploidy, 69.6%) than in Group B (DNA aneuploidy and DNA peridiploidy, 33.2%; P < 0.001). The survival curves of the patients in Group B continued to decrease during the next 2.5 years. Coxs model analysis showed that both the pathologic stage and the DNA content were the significant prognostic factors for survival. However, the DNA content was an independent prognostic factor in squamous cell carcinoma, but not in adenocarcinoma. These results indicate that DNA content analysis is useful for the evaluation of clinical behavior and prognosis, and that the clinical value of the DNA content must be differentiated between squamous cell carcinoma and adenocarcinoma.

The Relationship of the Proliferating Cell Nuclear Antigen Protein to cis-Diamminedichloroplatinum(II) Resistance of a Murine Leukemia Cell Line P388/CDDP

We investigated whether the proliferating cell nuclear antigen (PCNA) protein takes part in cis-diamminedichloroplatinum (II) (CDDP) resistance, using a murine leukemia cell line P388 and its CDDP resistant cell line. P388/CDDP was 4 times more resistant to CDDP than P388. The cell lines were maintained in the DBA/2 female mouse peritoneal space. In total cells, the amount of the PCNA protein decreased to 90% in P388 after 1 h CDDP treatment, but that of P388/CDDP increased to 127%. The difference was statistically significant (p = 0.012, n = 5). As for G2/M phase cells, the difference was also significant at 1 h (p = 0.016, n = 5) and at 2 h (p = 0.036, n = 5) after treatment. In P388 the amount of the PCNA protein decreased in accordance with the inhibited cell proliferation, whereas in P388/CDDP, the amount of the PCNA protein increased in spite of the inhibited cell proliferation. This increase of the PCNA protein suggests that the PCNA protein is involved in CDDP resistance of P388/CDDP through enhanced DNA repair synthesis.

Accuracy of the bronchoscopic DNA content analysis of non-small-cell lung carcinoma

To assess the accuracy of the bronchoscopic DNA content analysis, samples of non-small-cell lung carcinomas (NSCLC) were investigated by means of flow cytometry. Samples were dissociated using the detergent Triton X-100. In 58 NSCLC cases, 39 (67%) had DNA aneuploid tumors. We compared the DNA indices of bronchoscopic brushing samples with 21 corresponding surgical samples. In 16 (76%) cases, DNA ploidy of both bronchoscopic and surgical samples were in concordance. In 3 (14%) cases, both bronchoscopic and surgical sample showed DNA aneuploidy, but the number of the DNA aneuploid stem cell lines was different. The cause of these differences was ascribed to the intratumor DNA heterogeneity. In 2 (10%) cases, the bronchoscopic sample showed DNA diploidy, but the surgical sample showed DNA aneuploidy. In these cases, tumor cells obtained by bronchoscopic brushing were so few that the small DNA aneuploid peak was undetectable in the DNA histogram. But the tumor DNA ploidy was evaluated correctly in 90% of 21 cases using bronchoscopic samples. Consequently, despite some drawbacks, the DNA ploidy diagnosis using bronchoscopic samples in this relatively small study, was almost as reliable as surgical samples.

Nuclear DNA Content and Expression of ras Oncogene p21 in Non-Small Cell Lung Cancer.

切除肺癌の核DNA量を測定し, また, ras癌遺伝子産物p21蛋白の発現を免疫組織化学的に調べ, それらの肺癌患者の予後における意義を検討した.DNA量を測定した84腫瘍のうち, 19例 (23%) がDNA diploid腫瘍であり, 65例 (77%) がaneuploid腫瘍であった.rasp21の染色性をみた91例のうち, 58例 (64%) が陽性 (weak+strong腫瘍) であった.DNAdiploid腫瘍患者の5年生存率は61%, aneuploid腫瘍患者は35%であり, aneuploid腫瘍患者の予後は有意に不良であった (p<0.01).p21-negative腫瘍患者の5年生存率は64%, p21-weakは38%, p21-strongは12%で発現性が強い症例ほど予後は不良であった.病期, 核DNA ploidy, ras p21発現性の3因子を組み合わせて, Coxの重回帰型生命表を用い, Risk scoreを算出した.Risk score3.0以下を低危険群, 3.1から5.0までを中危険群, 5.1以上を高危険群の三群に分類すると, 低危険群の5年生存率は100%と全員が生存しており, Riskscoreが大きくなるほど予後が不良であった (p<0.0001).以上より, 予後リスク分類は肺非小細胞癌患者の術後予後の推定に有用である.

Differences of nuclear DNA content between central and peripheral squamous cell carcinoma of the lung.

切除された肺扁平上皮癌48例の核DNA量をフローサイトメトリーで測定し, 中心型と末梢型腫瘍に分けて検討した. 中心型腫瘍 (23例) のDNAaneuploidyの頻度は82.6%, DNAIndex (DI) は1.65±0.40 (±SD) であった. 一方, 末梢型腫瘍 (25例) ではそれぞれ40.0%, 1.29±0.37で末梢腫瘍の方がaneuploidyの頻度は少なく, DIは小さい値を示した (P<0.01). 中心型より末梢型腫瘍の方が, aneuploidよりdiploid腫瘍の方が予後は良かった (それぞれ, p<0.01). 女性6例 (非喫煙者5例) 全例が末梢型腫瘍であり, 且つ, diploid腫瘍であった. 核DNA量の測定は中心型と末梢型扁平上皮癌の悪性度や病因の違いを明らかにする手段として有用である.

Flow cytometric analysis of DNA index in lung adenocarcinoma: Comparison with grade of differentiation, histological subtypes and cellular subtypes.

原発性肺腺癌の手術患者go例を分化度別, 組織亜型別および細胞亜型別に分類し, それらと核DNA量, 患者予後との関連性について検討した. DNA aneuploidyは90例中, 72例 (80%) に認められ, DNA index (DI) のmean±SDは1.76±0.56であった. 核DNA量は肺胞上皮型で少ない傾向であったが有意差はなかった. 患者予後は, 分化度別で高分化型が, また組織亜型分類で肺胞上皮型が良好であった. Cuboidal cell type (立方状型) の患者群では, 他の群よりも早期からリンパ節転移をおこしやすい傾向であった. Columnar cell type without mucus (粘液を含まない高円柱状型) では核DNA量が他の群よりも大きく (平均DI=2.25), またsmoking indexが平均1,300と高かった. 細胞亜型別にDIを比較すると, smoking indexの分布と同じ傾向を示し, 喫煙が一部の肺腺癌の発生や核DNA量に影響を及ぼしている可能性がある.