Yukashi Ito

DNA methylation analysis at distal and proximal promoter regions of the oestrogen receptor gene in breast cancers

SummaryOestrogen receptor α (ER-α) gene has two specific promoters, distal (P0) and proximal (P1), which induce almost identical transcripts in size due to different splicing. We examined the methylation at both promoter regions of the ER-α gene using HpaII, a methylation-sensitive restriction enzyme, prior to polymerase chain reaction (PCR) amplification. To confirm the results of PCR-based methylation analysis, Southern hybridization was also performed. Twenty of 29 patients with ER-α-positive tumours and five of 27 with ER-α-negative tumours were unmethylated at the P1 promoter region of the ER-α gene. The incidence of methylation was highly negatively correlated with ER-α expression (P = 0.0002). A similarly negative correlation was observed at the P0 promoter region of the ER-α gene (P = 0.0154). Additionally, the tumours with the ER-α gene hypermethylated at both promoter regions had definitely negative ER-α values. It was suggested that this epigenetic change might control ER-α expression, and might play an important role in the loss of hormone-dependence in breast cancer.

Predictors of response to exemestane as primary endocrine therapy in estrogen receptor–positive breast cancer

Endocrine therapy is the most important treatment of choice for estrogen receptor (ER)‐positive breast cancer. Potential mechanisms for resistance to endocrine therapy involve ER‐coregulatory proteins and cross‐talk between ER and other growth factor–signaling networks. However, the factors and pathways responsible for endocrine therapy resistance, particularly resistance to aromatase inhibitors, have not been clearly established. Sixteen postmenopausal patients with ERα‐positive primary breast cancer were treated daily with 25 mg of exemestane (an aromatase inhibitor) for 6 months. Expressions of ERα, ERβ, progesterone receptor (PgR), androgen receptor (AR), amplified in breast cancer 1 (AIB1), aromatase, epidermal growth factor receptor, human epidermal growth factor receptor type 2, Ki67, cyclin D1, p53, Bcl2, signal transducer and activator of transcription 5 (Stat5), and insulin‐like growth factor binding protein 5 (IGFBP5), and phosphorylations of ERα serine (Ser) 118, ERα Ser167, Akt Ser473, and p44/42 MAPK threonine (Thr) 202/tyrosine (Tyr) 204, were examined by immunohistochemistry on pretreatment tumor biopsies and post‐treatment surgical specimens. Analyses were made to test for correlations with response to exemestane. Of the 16 patients, seven responded and nine retained stable disease. High‐level expression of AIB1 and phosphorylation of Akt Ser473 were significantly associated with a better response to exemestane, suggesting that these factors could be considered as predictors of exemestane response. Expressions of ERα, ERβ, PgR, aromatase, Ki67, cyclin D1, and p53, and phosphorylations of ERα Ser118, ERα Ser167, and p44/42 MAPK Thr202/Tyr204, were decreased, whereas expressions of Stat5 and IGFBP5 were increased in post‐treatment specimens compared to the values in pretreatment biopsies. Thus, the analysis of factors involved in the estrogen‐dependent growth‐signaling pathways may be useful in identifying patients responsive to exemestane. (Cancer Sci 2009)

Clinical significance of bcl-2 gene expression in human breast cancer tissues

The expression of estrogen receptor (ER) and bcl-2(Bcl-2), an apoptosis protective oncogene, in normal andcancerous breast duct epithelia was immunohistochemically examined infresh frozen tumor tissues from 142 Japanese breastcancer patients. The clinico-pathological characteristics and the diseasefree survival of the patients were analyzed. Theexpression of both the proteins was also observedin intraductal components of breast cancer. Although lessthan 1% of normal duct epithelia expressed ER,Bcl-2 was diffusely expressed. The expression of boththese proteins in breast cancer significantly correlated witheach other. Their expression significantly correlated negatively withtumor size but not with lymph node status.The papillo-tubular sub-type of invasive ductal carcinoma expressedBcl-2 significantly more frequently than the solid-tubular sub-type.Patients with Bcl-2 expressing tumors survived without recurrencesignificantly more than those with tumors exhibiting reducedexpression. Papillary-cribriform type intraductal componentsexpressed both those proteins more often than the solid-comedo type.

Immunohistochemical analysis on biological markers in ductal carcinoma in situ of the breast

BackgroundThe increasing use of mammographic screening has led to an increased detection of ductal carcinomain situ (DCIS) of the breast. The detailed biological characteristics of DCIS and a new classification of DCIS based on these characteristics are needed.MethodsImmunohistochemical studies were performed to assess the expression of c-erbB-2 (ErbB-2), estrogen receptor (ER), p53 and proliferative activity (Ki-67) in 65 patients with pure DCIS and 60 with invasive ductal carcinoma (IDC). We classified pure DCIS tumors using three classifications, the architectural, Nottingham, and Van Nuys classifications.ResultsErbB-2, ER and p53 staining was positive in 34%, 66% and 21% of patients with DCIS, respectively, and 58%, 42% and 33% in patients with IDC, respectively. Ki-67 stained positively in 1.5 % of patients with DCIS and 11.2 % of patients with IDC. The comedo type showed a high rate of positive ErbB-2 and p53 staining. The cribriform and papillary types showed a high rate of positive ER staining. Under the Van Nuys classification, ErbB-2, p53 and Ki-67 expression were highest in the group with high nuclear grade and lowest in the group with non-high nuclear grade without necrosis.ConclusionAlthough the biological markers of IDC tended to suggest aggressive behavior more so than those of DCIS, these differences were based on the histological sub-type, comedo or non-comedo. The Van Nuys classification best defined the subgroups of DCIS with a distinct expression pattern of biological markers, and the best candidates for breast-conserving surgery.

Comparison of five different antibodies in the immunohistochemical assay of estrogen receptor α in human breast cancer

BackgroundEstrogen receptor α(ER) expression is the best prognostic and predictive factor of hormone dependency of human breast cancers. Unlike enzyme immunoassay (EIA), which has been widely used to evaluate ER status in breast cancer, immunohistochemical assay (IHC) can detect ER in a small amounts of tissue with detailed localization. Although there is a sufficient number of ER antibodies against various regions of the protein, the reliability of IHC staining is only well understood for a few. IHC and EIA for the evaluation of the ER status of human breast cancer, therefore, should be compared using the same breast cancer tissues.MethodsFive different ER antibodies (1D-5, C-314, G-20, C-311 and HC-20) that identify different amino acid sequences were used. The evaluation of ER status by IHC using these antibodies was compared with EIA concomitantly in 97 primary human breast cancer tissues.ResultsThe positiviry rate for EIA was 68%. That of IHC for antibodies 1D-5, C-314, G-20, C-311 and HC-20 was 50.5%, 47.4%, 46.4%, 44.3% and 57.7%, respectively. The concordance between EIA was 76.3% for 1 D-5 and 77.3% for HC-20, which is statistically highly significant (p<0.0001); Other antibodies were not.ConclusionsHC-20 is most suitable in the evaluation of the ER status of human breast cancers using the IHC method. Although antibody 1 D-5 is also available, C-314, G20 and C-311 are unreliable in such an evaluation.

Metachronous Primary Hyperparathyroidism Due to a Parathyroid Adenoma and a Subsequent Carcinoma: Report of a Case

Abstract An extremely rare case of metachronous primary hyperparathyroidism (PHP) due to a parathyroid adenoma and subsequent carcinoma with local lymphatic spread is presented herein. A 55-year-old woman was operated on for a parathyroid adenoma in the right inferior gland. Thirteen years after the first operation, she was again hospitalized for hypercalcemia and the presence of a hard mass in the right anterior neck region. Exploratory surgery and a histological examination of the resected tumor provided evidence of a parathyroid neoplasm in the right superior gland but the malignancy was equivocal. Postoperatively, her serum parathyroid hormone (PTH) level remained at 1.5-fold the upper limit of the normal range, and hypercalcemia again gradually developed. The results of higher positive rates by Ki-67 immunohistochemical staining and an aneuploid pattern by a flow cytometric analysis of the second neoplasm were consistent with a histological diagnosis of carcinoma, and she therefore underwent further surgery. A radical neck dissection revealed two lymph node metastases which were both successfully removed. The postoperative serum PTH and calcium levels then returned to within the normal ranges. These findings indicate the usefulness of Ki-67 immunohistochemical staining and a flow cytometric analysis for differentiating malignant lesions from benign parathyroid lesions, and the importance of surgically treating cases limited to local regions without distant metastasis.

Predictors of Response to Exemestane as Primary Endocrine Therapy in Estrogen Receptor-Positive Breast Cancer.

Endocrine therapy is the most important treatment of choice for estrogen receptor-positive breast cancer. Potential mechanisms for resistance to endocrine therapy involve estrogen receptor (ER)-coregulatory proteins and cross-talk between ER and other growth factor-signaling networks. However, the factors and pathways responsible for endocrine therapy resistance, particularly resistance to aromatase inhibitors, have not been clearly established. Sixteen postmenopausal patients with ERΑ-positive primary breast cancer were treated daily with 25 mg of exemestane (an aromatase inhibitor) for 6 months. Expressions of ERΑ, ERΒ, progesterone receptor (PgR), androgen receptor (AR), AIB1, aromatase, EGFR, HER2, Ki67, cyclin D1, p53, Bcl2, Stat5 and IGFBP5, and phosphorylations of ERΑ serine (Ser) 118, ERΑ Ser167, Akt Ser473 and p44/42 MAPK threonine (Thr) 202/ tyrosine (Tyr) 204 were examined by immunohistochemistry on pretreatment tumor biopsies and posttreatment surgical specimens. Analyses were made to test for correlations with response to exemestane. Of the 16 patients, 7 responded and 9 retained stable disease. High-level expression of AIB1 and phosphorylation of Akt Ser473 were significantly associated with a better response to exemestane, suggesting that these factors could be considered as predictors of exemestane response. Expressions of ERΑ, ERΒ, PgR, aromatase, Ki67, cyclin D1 and p53, and phosphorylations of ERΑ Ser118, ERΑ Ser167 and p44/42 MAPK Thr202/Tyr204 were decreased, whereas expressions of Stat5 and IGFBP5 were increased in posttreatment specimens compared to the values in pretreatment biopsies. Thus, the analysis of factors involved in the estrogen-dependent growth-signaling pathways may be useful in identifying patients responsive to exemestane. Citation Information: Cancer Res 2009;69(24 Suppl):Abstract nr 2041.

Primary adrenocortical tumors in autopsy records —A survey of “cumulative reports in Japan” from 1973 to 1984—

All cases of primary adrenocortical tumors, including those found incidentally, recorded in the Pathological Autopsy Case Annuals of Japan during a 12 year period from 1973 to 1984 were surveyed. Age- and sex-specific incidences of lesions were estimated from the population, mortality and autopsy statistics for the same period. There were 101 adenomas, 77 of which were presumed to be non-functional, and 222 carcinomas. The age-specific distribution profiles of both the benign and malignant tumors showed single peaks in the sixties and could not be distinguished from one another. The estimated age-specific distribution of morbidity of the non-functional adenomas showed a peak at about 10 years younger than that found in the autopsy records, although it was again similar to that of carcinoma derived from various clinical reports. The morbidity of the non-functional adenomas per 100,000 population was almost equal to that of breast cancer in Japan, being highest in the fifties age group with more than 4,000 people a year being suggested to have adenoma in this age bracket.

A case of pancreatic ascites successfully treated with peritoneal lavage.

慢性膵炎に起因する膵性腹水の1例を報告した.患者は20年来アルコールを多飲していた39歳の男性で, 急激な腹部膨満と上腹部激痛で受診した.臨床所見と諸検査によって, 多量の膵性腹水とともに肝硬変と慢性膵炎が存在すると診断された.全身的療法の効果が乏しいことから, 発症16日目より腹膜灌流を開始した.メシル酸ガベキサート200mg, アミカシン100mgおよびヘパリン2,000単位を含む21の等張腹膜灌流液を腹腔内に60分で注入し, 引き続き120分で自然流出させた.この持続的腹膜灌流は3時間ごとに5日間連続しておこなった.この治療によって臨床症状, 血清アミラーゼをはじめとする検査結果は著しく改善した.すなわち, 蛋白分解酵素阻害剤を添加した等張の腹膜灌流液による連続的腹膜灌流力が有用であった.しかし, 腹膜灌流の時期と継続期間, 蛋白分解酵素阻害剤の種類と量, などなお考慮すべき点が残されている.

A case of superior mesenteric arterial occlusion successful treated with thrombectomy.

は じめ に 上 腸管膜 動脈 閉塞 症(以 下 本症)は 早期 診断 が困難 で,広 範 囲 な腸 壊死 に至 り開腹 され る こ とが多 く予後 は きわ めて不 良 とされ てい る1). わ れわ れ は腸切 除を 必要 とせず 血栓 除去 術 のみ で救 命 しえた70歳 男性 の1例 を経 験 した ので,当 院で経験 した5例 の本 症 と併せ て報 告す る. 症 例 患 者:70歳,男 性. 主 訴:上 腹 部痛. 家 族 歴,生 活歴:特 記す べ きこ とな し. 既 往 歴:昭 和61年9月 に心房 細動 と診 断 され,以 後 投薬 治 療 を うけてい る. 現病 歴:昭 和61年11月5日 午 後1時 半,突 然上 腹部 に激痛 を感 じ,食 物残 渣 を嘔 吐 した.た だ ちに救 急車 にて 当院緊 急入 院 とな った. 入 院 時 現 症:身 長165cm,体 重68kg,血 圧186/100 mmHg,脈 拍94/分 で不 整 であ り,眼 瞼結 膜 に貧血,眼 球 強膜 に黄 疸 を認め ず.腹 部 は軟 でやや 膨隆 して お り, 肝,脾,腫 瘤 を触知 しない.上 腹 部 に強 い 自発痛 と軽 度の圧 痛 を認 め るが,腹 膜刺 激症 状 は欠 いてい た.入 院2時 間 後 に は トマ トケチ ャ ップ様 の 血便 を 約300ml 認め た. 検査 成績:血 液生 化学 検査 で は 白血球 の軽 度 増多 お よび軽 度 の脱 水 を 認 め る 以外 は ほ ぼ 正常 で,GOT, GPT,CPK,LDHも 異 常 は なか った(表1).心 電 図 で は心房 細動 を認 め,腹 部 単純X線 写 真 では腸 管 ガ ス の増 加,鏡 面 像 な どの イ レウス所 見 は認 めな か った. 腹 部computed tomography(CT):腎 に嚢胞 を 腹 大動脈 に壁 の石灰 化を 認 めた.ま た確 定 診断後 に検 討