Anti-cancer stem cell activity of the Src inhibitor dasatinib in thyroid cancer cells

Although the prognosis of differentiated thyroid cancer (DTC) is good, those of poorly-differentiated and undifferentiated thyroid cancers (PDTC and UDTC) are poor. Recent preclinical studies have suggested that the Src inhibitor dasatinib is active in thyroid cancer cell lines. We conducted the present study in an attempt to clarify the antitumor activity of dasatinib in PDTC and UDTC. The expression levels of c-Src, phosphorylated Srcs (p-SrcY416 and p-SrcY527), focal adhesion kinase (FAK), and phosphorylated FAK (p-FAKY861) were immunohistochemically investigated in a case-control series (15 cases of PDTC or UDTC vs. 29 control cases of DTC). The PDTC cell line KTC-1 and UDTC cell line KTC-2 were used to investigate the anticell growth and anti-cancer stem cell (CSC) activities of dasatinib. The combined effects of dasatinib and the taxane paclitaxel on anti-cell growth and anti-CSC activities were also tested. c-Src and p-FAKY861 expression levels were significantly higher, while those of p-SrcY416 were slightly higher in PDTC and UDTC than in DTC. Dasatinib inhibited cell growth in association with G1-S cell cycle retardation and increased apoptosis in both cell lines. Dasatinib significantly decreased the proportion of CSCs and more than additively enhanced the anti-cell growth activity of paclitaxel. The results of this study suggest that the Src signaling pathway is activated more in PDTC and UDTC than in DTC. The Src inhibitor dasatinib exhibited anti-cell growth and anti-CSC activities. Furthermore, it more than additively enhanced the anti-cell growth activity of paclitaxel

World squid fisheries

Peer reviewedPublisher PD

Genotype determination of the OPN1LW/OPN1MW genes: novel disease-causing mechanisms in Japanese patients with blue cone monochromacy

Blue cone monochromacy (BCM) is characterized by loss of function of both OPN1LW (the first) and OPN1MW (the downstream) genes on the X chromosome. The purpose of this study was to investigate the first and downstream genes in the OPN1LW/OPN1MW array in four unrelated Japanese males with BCM. In Case 1, only one gene was present. Abnormalities were found in the promoter, which had a mixed unique profile of first and downstream gene promoters and a −71A > C substitution. As the promoter was active in the reporter assay, the cause of BCM remains unclear. In Case 2, the same novel mutation, M273K, was present in exon 5 of both genes in a two-gene array. The mutant pigments showed no absorbance at any of the wavelengths tested, suggesting that the mutation causes pigment dysfunction. Case 3 had a large deletion including the locus control region and entire first gene. Case 4 also had a large deletion involving exons 2–6 of the first gene. As an intact LCR was present upstream and one apparently normal downstream gene was present, BCM in Case 4 was not ascribed solely to the deletion. The deletions in Cases 3 and 4 were considered to have been caused by non-homologous recombination

Frequently increased epidermal growth factor receptor (EGFR) copy numbers and decreased BRCA1 mRNA expression in Japanese triple-negative breast cancers

<p>Abstract</p> <p>Background</p> <p>Triple-negative breast cancer (estrogen receptor-, progesterone receptor-, and HER2-negative) (TNBC) is a high risk breast cancer that lacks specific therapy targeting these proteins.</p> <p>Methods</p> <p>We studied 969 consecutive Japanese patients diagnosed with invasive breast cancer from January 1981 to December 2003, and selected TNBCs based on the immunohistochemical data. Analyses of epidermal growth factor receptor (<it>EGFR</it>) gene mutations and amplification, and <it>BRCA</it>1 mRNA expression were performed on these samples using TaqMan PCR assays. The prognostic significance of TNBCs was also explored. Median follow-up was 8.3 years.</p> <p>Results</p> <p>A total of 110 (11.3%) patients had TNBCs in our series. Genotyping of the <it>EGFR </it>gene was performed to detect 14 known <it>EGFR </it>mutations, but none was identified. However, <it>EGFR </it>gene copy number was increased in 21% of TNBCs, while only 2% of ER- and PgR-positive, HER2-negative tumors showed slightly increased <it>EGFR </it>gene copy numbers. Thirty-one percent of TNBCs stained positive for EGFR protein by immunohistochemistry. <it>BRCA1 </it>mRNA expression was also decreased in TNBCs compared with controls. Triple negativity was significantly associated with grade 3 tumors, TP53 protein accumulation, and high Ki67 expression. TNBC patients had shorter disease-free survival than non-TNBC in node-negative breast cancers.</p> <p>Conclusion</p> <p>TNBCs have an aggressive clinical course, and <it>EGFR </it>and <it>BRCA1 </it>might be candidate therapeutic targets in this disease.</p

ベバシズマブ併用化学療法中に消化管穿孔をきたした再発乳癌の１例

　ベバシズマブはパクリタキセルとの併用でHER2陰性の進行・再発乳癌に対する有効性が示されており，無増悪生存期間を有意に延長させる．しかし，ベバシズマブ特有の有害事象も報告されており，投与の際には注意を要する．今回，再発乳癌に対しベバシズマブを使用し，腸管穿孔を起こした１例を経験した．症例は72歳女性．右乳癌術後５年目に多発リンパ節，肺転移を認め，化学療法で治療中に８次治療としてベバシズマブとパクリタキセル（BP）療法を開始した．１年ほど奏効したが，突然，腹痛を訴え受診した．CT で腹腔内にfree air を認めたため緊急開腹術を施行した．小腸に１か所の穿孔部位を認めた．病理組織検査では，穿孔部に乳癌の転移巣が認められた．乳癌に対するベバシズマブ併用化学療法中の消化管穿孔は報告が少ない．腹膜播種を認める症例やベバシズマブ投与期間の長い患者では，腹部膨満感や腹痛を訴えた際は消化管穿孔を念頭におく必要がある．　Combination therapy with bevacizumab and paclitaxel (BP therapy) has been reported to be effective for the treatment of HER2-negative metastatic breast cancer and to significantly prolong progression-free survival. However, there are specific adverse effects induced by bevacizumab that physicians should pay attention to. We report a recent case of metastatic breast cancer with gastrointestinal perforation during bevacizumab therapy. A 72-year-old female patient had metastases into multiple lymph nodes and lungs five years after surgery for primary breast cancer, and was treated with several chemotherapies. The patient received BP therapy as the eighth treatment regimen. Although the therapy led to stable disease for approximately one year, the patient suddenly developed abdominal pain. Emergency laparotomy was performed because computed tomography revealed free air in the peritoneal cavity. A perforated lesion was found in her small intestine. On pathological examination, breast cancer metastasis was noted around the perforated site. There are few reports of gastrointestinal perforation during bevacizumab therapy for patients with metastatic breast cancer. When a patient has peritoneal dissemination, long-term BP therapy and abdominal pain, physicians should keep in mind the possibility of gastrointestinal perforation during BP therapy. (187 words

術後化学療法後に持続性無月経であったが高エストロゲン血症を呈した閉経前乳癌の1症例

ホルモン感受性乳癌患者においては,内分泌療法を選択する上で,閉経状況が重要である.また,先行する化学療法によって無月経になることがあり,閉経前・後の判断は難しい.今回,我々は化学療法後に無月経状態であったにも関わらず,高エストロゲン血症を呈した1症例を経験したので報告する.症例は診断時42歳の女性.左乳癌に対して左乳房切除及び腋窩リンパ節郭清術を施行した.病理検査結果は硬癌,核グレードIII,エストロゲン受容体陽性,プロゲステロン受容体陽性,HER2陰性,リンパ節転移2個であり,術後補助療法として複合化学療法施行後にタモキシフェンを内服していた.化学療法中より無月経であったが,化学療法開始後2年4ヶ月後にホルモン状態を確認したところ血清エストラジオール(E2)は567.2 pg/mlと高値であった.化学療法後に1年以上無月経であっても,卵巣機能は保持されている症例があり,定期的な血清中のE2およびFSH を測定し,閉経状況を評価する必要がある.In patients with endocrine-sensitive breast cancer, the menopausal status is important for the selection of endocrine therapy. When chemotherapy is administered prior to endocrine therapy, it becomes more difficult to judge the menopausal status. We report a premenopausal breast cancer patient who developed amenorrhea after postoperative adjuvant chemotherapy despite a high serum estrogen level. The patient was a 42-year-old woman diagnosed with left breast cancer. She underwent left mastectomy and axillary lymph node dissection. The pathological findings revealed scirrhous carcinoma, which was estrogen receptor-positive, progesterone receptor-positive, and HER2-negative, with two positive lymph nodes. She received tamoxifen after combined cytotoxic chemotherapy as postoperative adjuvant therapy. She developed amenorrhea during the chemotherapy, which continued for two years and four months. Her serum estradiol (E2) level was very high (562.2 pg/ml). After further examinations, she was diagnosed with hypothalamuspituitary gland-related amenorrhea. Even if amenorrhea persists over a year after chemotherapy, the ovarian function might be preserved, as in this case. Therefore, serum levels of E2 and FSH should be periodically measured during endocrine therapy

乳腺神経内分泌癌の一例

稀な癌である乳腺神経内分泌癌(neuroendocrine carcinoma:以下NEC)を経験したので報告する.患者は35歳の女性で,右乳頭直下に腫瘤を自覚し当科を受診した.画像診断で乳癌を疑い,細胞診では乳頭腺管癌あるいは硬癌が推定された.針生検で浸潤性乳管癌と診断し,乳房温存術とセンチネルリンパ節生検を施行した.腫瘍は乳腺原発であり,組織像は,小細胞癌などの神経内分泌癌に類似していた.免疫組織化学的検査では,chromogranin A, synaptophysin, neuronspecific enolase陽性細胞が大部分を占めていたためWHO分類に従い神経内分泌癌の診断に至った.リンパ節転移は認めなかった.乳腺NECは稀な癌であり,臨床病理学的な特徴,予後,治療法についての定まった見解はない.本症例では,通常の乳癌同様に臨床病理学的予後因子に基づき術後補助化学療法を行った.術後8年5カ月の時点で無再発生存中である.We are reporting a rare case of breast neuroendocrine carcinoma (NEC). A 35-year- old woman noticed a breast lump underneath her right nipple, and she consulted our hospital. The mass was suspected of being breast cancer by mammography, ultrasonography and MRI. Cytological diagnosis was papillotubular or scirrhous carcinoma. Finally, it was diagnosed as invasive ductal carcinoma by a core needle biopsy. We performed breastconserving surgery and sentinel lymph node biopsy. There was no metastasis in the sentinel lymph node. Pathological findings suggested that it was a small cell carcinoma or a neuroendocrine carcinoma. Chromogranin A, synaptophysin, neuron-specific enolase were positively stained in most of the cancer cells by immunohistochemistry. Therefore, it was diagnosed as breast NEC according to the WHO classification. Because breast NEC is a relatively rare cancer, its clinicopathological significance, prognosis and appropriate treatment has not been evaluated. We chose the adjuvant systemic therapy based on recommendations from the St.Gallen Consensus Conference in this case. 101 months after the operation, she has no recurrence and is alive

転移・再発乳癌患者に対するエリブリン療法の有用性

　エリブリンはタキサンとは異なる作用機序をもつ微小管阻害剤である．海外の第Ⅲ相試験 では，エリブリンの転移・再発乳癌に対する延命効果が示されている．今回，エリブリンの臨床 的な有用性を検討するため，2011年９月から2017年８月に当科でエリブリン療法を行った進行・ 再発乳癌97症例を対象として後方視的に調査した．対象患者の年齢は35 － 81歳（中央値58）， performance status は１が最多で64例，Stage Ⅳが５例，再発が92例であった．原発腫瘍のエス トロゲン受容体は陽性が64例，プロゲステロン受容体は陽性が48例，human epidermal growth factor receptor 2は陰性が78例であった．前化学療法のレジメン数は0 － 9（中央値２）, 臓器転 移ありが69例，肝転移ありが40例，エリブリン療法の実施サイクル数は1 － 12回（中央値3.5）， 観察期間は1 － 55か月（中央値10），有害事象による中止例は10例であった．最大治療効果は，完 全奏効が０例，部分奏効が１例，長期安定が27例，安定が16例，進行が42例，不明・評価不能が 11例であった． 臨床的有効率（奏効率＋長期安定率）は29% であった．Time-to-treatment failure (TTF) は0 － 178週間（中央値13），治療開始後全生存期間は0 － 55か月（中央値15.5）であった． 好中球減少症はグレード１が最多で61例，非血液毒性は嘔気が７例，肝機能障害が６例，末梢神 経障害が５例，間質性肺炎が3例などであった．良好なTTF の予測因子は，単変量解析で「臓器転 移なし」（P = 0.0356）が同定された．良好な治療開始後生存の予測因子は，多変量解析にて「臨 床的有効性あり」（P = 0.0008）と「PS が０か１」（P < 0.0001）が同定された．エリブリン療法は， 奏効率は低かったが，本療法は，約30% の症例に臨床的有効性をもたらし，生存期間の延長に寄 与する可能性がある．　Eribulin is an anti-microtubule agent that uses a different mechanism of action to taxanes. Phase 3 clinical trials have shown that eribulin exerts life-prolonging effects in patients with metastatic or recurrent breast cancer. In order to investigate the utility of eribulin, we conducted a retrospective and observational study on 97 patients with metastatic and recurrent breast cancer who were treated in our institute between September 2011 and August 2017. The median age of the patients was 58 years (range: 35 － 81). The performance status was 1 in 64 patients. Five patients had stage IV disease, while 92 had recurrent disease. Sixtyfour patients had estrogen receptor-positive tumors, 48 had progesterone receptor-positive tumors, and 78 had human epidermal growth factor receptor 2-negative tumors. The median number of regimens of previous chemotherapies was 2 (range: 0 － 9). Sixty-nine patients had visceral metastases, while 40 had liver metastases. The median number of cycles of eribulin therapy was 3.5 (range: 1 － 12). The median follow-up period was 10 months (range: 1 － 55). The best responses to therapy were a complete response in 0 patients, a partial response in 1, long-term stable disease in 27, stable disease in 16, progressive disease in 42, and unevaluable in 11. The clinical benefit rate (objective response rate + long-term stable disease rate) was 32%. The median time-to-treatment failure (TTF) was 13 weeks (range: 0 － 178). Median overall survival (OS) after the initiation of therapy was 15.5 months (range 0 － 55). The most frequent grade of neutropenia was 1, which was observed in 61 patients. Major non-hematological toxicities were nausea in 7 patients, liver dysfunction in 6, peripheral neuropathy in 5, and interstitial pneumonitis in 3. Univariate and multivariate analyses identified “no visceral metastasis” as the only predictive factor for TTF (P = 0.0356 for the multivariate analysis). The multivariate analysis revealed that“ the presence of a clinical benefit by the therapy” (P = 0.0008) and “a performance status of 0 or 1” (P < 0.0001) were independent predictive factors for OS. Eribulin therapy for patients with metastatic or recurrent diseases provided clinical benefits for approximately 30% of patients. These results suggest that this therapy prolongs the OS of patients

進行・再発乳癌患者に対するフルベストラントの有用性

　ホルモン受容体陽性の閉経後進行・再発乳癌患者に対する内分泌療法として，フルベストラントが本邦で臨床導入されて６年余りが経過した．本剤の有用性を検討するため，2012年１月〜2016年10月に川崎医科大学附属病院乳腺甲状腺外科において，フルベストラントが単独使用され，治療評価が可能であった51症例の電子カルテを後方視的に調査した．対象患者の年齢の中央値は70歳．進行例が９例，再発例が42例．臓器転移ありが23例．観察期間の中央値は18か月．前内分泌療法数の中央値は２．前化学療法歴ありは21例．治療効果は，完全奏効が３例，部分奏効が６例，安定が25 例（うち長期安定は20例），進行が16例であった．客観的奏効は９例（17.6%），臨床的有用は29例（56.9%）であった．無増悪生存（PFS）期間の中央値は８か月，全生存（OS）期間の中央値は34か月であった．治療効果の予測因子を調べるため，サブグループに分けてPFS及びOS を解析した．肝転移の有無では，PFS 期間の中央値は，なしが9.5か月，ありが５か月（P= 0.0386），OS 期間の中央値は，なしが41か月，ありが15か月（P = 0.0036）であった．前化学療法の有無では，PFS 期間の中央値は，なしが12.5か月，ありが3.5か月（P < 0.0001），OS 期間の中央値は，なしが41か月，ありが24か月（P = 0.0208）．多変量解析では，前化学療法歴の有無が唯一のPFS の有意な予測因子であった．また，肝転移の有無が唯一のOS の有意な予測因子であった．有害事象は６例（11.7%）に認めたが，いずれも軽微であり治療が中断されることはなかった．要約すると，ホルモン受容体陽性の進行・再発乳癌の治療薬として，フルベストラントは17.6% の客観的奏効率，56.9% の臨床的有用性が認められ，既知の報告と同等の治療効果であった．フルベストラントは，化学療法歴のある症例，肝転移のある症例では，有用性が低いと考えられた．　Fulvestrant has been used for the treatment of postmenopausal patients with advanced or recurrent breast cancer in Japan for over six years. To investigate the utility of fulvestrant, we retrospectively reviewed electronic medical records and evaluated the responses of 51 patients with advanced or recurrent breast cancer treated with fulvestrant alone at the Department of Breast and Thyroid Surgery, Kawasaki Medical School Hospital between January 2001 and December 2016. The median age of the subjects was 70 years old. Nine had stage IV diseases and 42 had recurrent diseases. Twenty-three patients had visceral metastases. The median follow-up time after the start of fulvestrant treatment was 18 months. The median number of previous endocrine therapies was 2. Twenty-one patients received chemotherapy previously. Three patients had a complete response, six had a partial response, 25 had a stable disease including 20 patients with a long-term stable disease, and 16 had progressive disease. The objective response rate was 17.6% (9 out of 51), and the clinical benefit rate was 56.9% (29 out of 51). The median progression-free survival (PFS) time was 8 months. The median overall survival (OS) time was 34 months. To investigate predictive factors for response to fulvestrant, subgroup analyses were performed. For liver metastasis, the median PFS and OS time were 9.5 and 41.0 months, respectively, in patients without liver metastasis but 5.0 and 15.0 months, respectively, in those with liver metastasis (P = 0.0386 and P = 0.0036, respectively). For previous chemotherapy, the median PFS and OS time were 12.5 and 41.0 months, respectively, in patients without previous chemotherapy but 3.5 and 24.0 months, respectively, in those with previous chemotherapy (P < 0.0001 and P = 0.0208, respectively). In addition, the Cox’s proportional hazards model revealed that previous chemotherapy was only an independent predictive factor for PFS and that liver metastasis was only an independent predictive factor for worse OS. Although toxicities were recorded in 6 of 51 patients (11.7%), all instances were slight and no patient stopped fulvestrant therapy because of toxicities. In summary, fulvestrant therapy at our hospital provided a 17.6% objective response rate and 56.9% clinical benefit rate in patients with advanced or recurrent breast cancer. These results were similar to those reported previously. According to our subgroup analyses, fulvestrant was unlikely to be effective in patients who received previous chemotherapy or had liver metastasis