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Kazuya Shinmura,
Hisaki Igarashi,
Masanori Goto,
Hong Tao,
Hidetaka Yamada,
Shun Matsuura,
Mari Tajima,
Tomonari Matsuda,
Arito Yamane,
Kazuhito Funai, Masayuki Tanahashi,
Hiroshi Niwa,
Hiroshi Ogawa,
Haruhiko Sugimura
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ABSTRACT: Activation-induced cytidine deaminase (AID) is expressed in B lymphocytes and triggers antibody diversification. Recent reports have indicated that the constitutive expression of AID in mice causes not only lymphomas, but also cancers of some organs including the lung, prompting us to investigate the expression and effect of AID on human lung cancer.
We examined AID mRNA expression in 17 lung cancer cell lines and 51 primary lung cancers using a quantitative RT-PCR analysis. Next, we established H1299 lung cancer cells stably overexpressing AID and performed a supF forward mutation assay. We then examined AID protein expression and p53 mutation in 129 primary lung cancers by an immunohistochemical analysis and PCR-SSCP and sequencing analyses, respectively.
Aberrant mRNA expression of AID was detected in 29% (5 of 17) of the lung cancer cell lines and 31% (16 of 51) of the primary lung cancers. AID-overexpressing H1299 clones showed a 5.0- to 6.1-fold higher mutation frequency than an empty vector-transfected H1299 clone, and about half of the AID-induced mutations were base substitutions, indicating that AID induces gene mutations in lung cancer cells. Furthermore, an association was found between the AID protein expression level and the p53 mutation status in an analysis of 129 primary lung cancers. A further expression analysis revealed that a portion of AID is localized at the centrosomes.
Our current findings suggest that the aberrant expression of AID may be involved in a subset of human lung cancers as a result of its mutation-inducing activity.
Annals of Surgical Oncology 02/2011; 18(7):2084-92. · 4.17 Impact Factor
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ABSTRACT: We analyzed 39 patients who underwent a 2nd resection for recurrent (solitary pulmonary metastasis) or 2nd primary lung cancer. Based on the pathological findings, 18 patients were diagnosed as recurrent lung cancer, and 21 patients were diagnosed as 2nd primary lung cancer. Overall 5-year survival was 69.4%. There are no difference between recurrent group and 2nd primary group. It is difficult to distinguish preoperatively between recurrent lung cancer and 2nd primary lung cancer, so we must be consider the 2nd resection as a curative resection for "2nd primary lung cancer".
Kyobu geka. The Japanese journal of thoracic surgery 10/2010; 63(11):944-9.
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ABSTRACT: Background: In this study, we analyzed the usefulness of adjuvant chemotherapy for non-small cell lung cancer based on the histoculture drug response assay (HDRA).
Methods: From September 2001 to December 2008, 65 patients with pathologic stage II or higher non-small cell lung cancer who underwent surgery received two-cycle HDRA-based adjuvant chemotherapy. Chemosensitivity to cisplatin, carboplatin, paclitaxel, docetaxel, gemcitabine, and irinotecan was examined by the HDRA. All patients were classified according to the number of administered HDRA-positive drugs: the prediction-sensitive group (PSG) (n = 31) comprised patients treated with two HDRA-positive drugs and the prediction-nonsensitive group (PNSG) (n = 34) comprised those treated with a combination of one HDRA-positive and one HDRA-negative drug or two HDRA-negative drugs. The clinical outcomes of the two groups were analyzed.
Results: The overall 5-year survival rate of the PSG was 82.4%. On the other hand, that of the PNSG was 40.1%. There were significant differences between the two groups (p = 0.03). The 5-year disease-free survival rate was more favorable in the PSG than in the PNSG (PSG: 56.5%, PNSG: 30.1%, p = 0.05). Multivariate analysis showed that chemotherapy based on the HDRA was a significant prognostic factor (p = 0.03).
Conclusions: The prognosis of patients treated with two HDRA-positive drugs was significantly better than that of those treated with one HDRA-positive drug or HDRA-negative drugs. Adjuvant chemotherapy based on the in vitro HDRA may be useful to improve survival in patients who have undergone surgery.
Journal of Thoracic Oncology 08/2010; 5(9):1376-1381. · 3.66 Impact Factor
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Kazuya Shinmura,
Hong Tao,
Kiyoko Nagura,
Masanori Goto,
Shun Matsuura,
Takahiro Mochizuki,
Kazuya Suzuki, Masayuki Tanahashi,
Hiroshi Niwa,
Hiroshi Ogawa,
Haruhiko Sugimura
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ABSTRACT: The candidate tumor suppressor NORE1A is a nucleocytoplasmic shuttling protein, and although a fraction of the NORE1A in cells is localized to their centrosomes, the role of centrosomal NORE1A has not been elucidated. In this study we investigated the role of NORE1A in the numerical integrity of centrosomes and chromosome stability in lung cancer cells. Exposure of p53-deficient H1299 lung cancer cell line to hydroxyurea (HU) resulted in abnormal centrosome amplification (to 3 or more centrosomes per cell) as determined by immunofluorescence analysis with anti-γ-tubulin antibody, and forced expression of wild-type NORE1A partially suppressed the centrosome amplification. The nuclear export signal (NES) mutant (L377A/L384A) of NORE1A did not localize to centrosomes and did not suppress the centrosome amplification induced by HU. Fluorescence in situ hybridization analyses with probes specific for chromosomes 2 and 16 showed that wild-type NORE1A, but not NES-mutant NORE1A, suppressed chromosome instability in HU-exposed H1299 cells that was likely to have resulted from centrosome amplification. We next examined the status of NORE1A mRNA expression in non-small cell lung carcinoma (NSCLC) and detected down-regulation of NORE1A mRNA expression in 25 (49%) of 51 primary NSCLCs by quantitative real-time-polymerase chain reaction analysis. These results suggest that NORE1A has activity that suppresses the centrosome amplification induced by HU and that NORE1A mRNA down-regulation is one of the common gene abnormalities in NSCLCs, both of which imply a key preventive role of NORE1A against the carcinogenesis of NSCLC.
Lung cancer (Amsterdam, Netherlands) 04/2010; 71(1):19-27. · 3.14 Impact Factor
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ABSTRACT: EML4-ALK fusion transcripts have been found in a subset of non-small cell lung carcinomas (NSCLCs); however, their protein expression status has not yet been fully elucidated. In this study we investigated ALK protein expression in 302 NSCLCs and 291 gastric carcinomas by means of immunohistochemical analysis. Twelve (4.0%) NSCLCs, but none of the gastric carcinomas, were found to be positive for ALK. The ALK signal was detected in the cytoplasm of cancer cells. Subsequent RNA analysis of 10 RNA-available, immunohistochemically ALK-positive tumors revealed that three tumors had EML4-ALK variant 1, three tumors had variant 2, three tumors had variants 3a and 3b, and one tumor had a novel variant in which exon 14 of EML4 is connected to the nucleotide at position 53 of exon 20 of ALK by a 2-bp insertion. These results suggest that immunohistochemical ALK detection is a useful way to screen NSCLCs for tumors containing ALK fusions.
Experimental and therapeutic medicine 01/2010; 1(2):271-275.
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Hiroshi Niwa, Masayuki Tanahashi,
Takashi Kondo,
Yoshinobu Ohsaki,
Yoshinori Okada,
Shigeki Sato,
Eiichi Suzuki,
Hiroshi Senba,
Shozo Fujino,
Teruomi Miyazawa,
Koichi Kobayashi
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ABSTRACT: In order to obtain information on the clinical application of bronchoscopy in Japan, the Japan Society for Respiratory Endoscopy (JSRE) conducted a postal survey.
A questionnaire was sent to 526 authorized institutes of the JSRE. The subject was bronchoscopy procedures performed during 2006.
The response rate was 71.3%. The total number of bronchoscopies performed was 74,770. Of these, 74,412 were flexible bronchoscopies and 358 were rigid bronchoscopies. At least one JSRE-authorized specialist had worked with 97% of respondents. Eighty-five per cent of respondents performed bronchoscopy under topical anaesthesia for almost all patients. Seventy-five per cent of respondents routinely used the oral route. The reported numbers of diagnostic bronchoscopies was 12,509 for simple bronchoscopy, 25,971 for forceps biopsy, 26,289 for brush biopsy, 25,659 for bronchial washing, 1387 for transbronchial needle aspiration and 6716 for BAL. Three deaths were caused by forceps biopsy (0.012%). The morbidity rates for these diagnostic procedures ranged from 0.14% to 2.5%. The reported numbers of therapeutic bronchoscopies was 476 for tracheobronchial stent, 164 for neodymium (Nd): yttrium-aluminium garnet (YAG) laser photoresection (LPR), 40 for photodynamic therapy, 81 for balloon dilatation, 145 for endobronchial electrocautery, 120 for argon plasma coagulation, 109 for microwave coagulation (MWC), 116 for ethanol injection, 110 for foreign body removal and 89 for bronchial occlusion. Deaths occurred only as a consequence of Nd : YAG LPR (0.61%). The morbidity rates for these therapeutic procedures ranged from 0% to 5%.
The preparation for, and practice of, bronchoscopy varied greatly between respondents. Diagnostic bronchoscopy was well tolerated and safe. Therapeutic procedures did not appear to be practised widely or frequently.
Respirology 02/2009; 14(2):282-9. · 2.42 Impact Factor
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Kazuya Shinmura,
Shinji Kageyama,
Hong Tao,
Tomoyasu Bunai,
Masaya Suzuki,
Takaharu Kamo,
Kazuya Takamochi,
Kazuya Suzuki, Masayuki Tanahashi,
Hiroshi Niwa,
Hiroshi Ogawa,
Haruhiko Sugimura
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ABSTRACT: EML4-ALK gene fusions have recently been discovered in a subset of human lung carcinomas, and fusions of the ALK tyrosine kinase gene with the NPM, TPM3, CLTC, ATIC, and TFG genes have been found in hematological malignancies. To elucidate the role of fusions between ALK and other genes in pulmonary carcinogenesis, we examined 77 non-small cell lung carcinomas (NSCLCs) for EML4-, NPM-, TPM3-, CLTC-, ATIC-, and TFG-ALK fusion transcripts by RT-PCR and subsequent sequencing analysis. Although no expression of NPM-, TPM3-, CLTC-, ATIC-, or TFG-ALK fusion transcripts were detected in any of the cases, expression of EML4-ALK fusion transcripts was detected in two (2.6%) of the 77 NSCLCs. In one of the two NSCLCs there was fusion between exon 13 of EML4 and exon 20 of ALK, i.e., variant 1, and in the other there was fusion between exon 20 of EML4 and exon 20 of ALK, i.e., variant 2. Both patients had a history of smoking, and histologically the carcinomas were adenocarcinoma. No somatic mutations were detected in the mutation cluster regions of the EGFR, K-RAS, and PIK3CA genes in these two carcinomas, however, a Pro177Ser mutation of the p53 gene was detected in the carcinoma that contained the variant 1 EML4-ALK fusion transcripts. In situ PCR of a paraffin block section showed that the carcinoma with expression of the variant 1 actually contained an EML4-ALK fusion gene. These results suggested that the EML4-ALK fusion gene product is involved in the carcinogenesis of a subset of NSCLCs.
Lung Cancer 02/2008; 61(2):163-9. · 3.43 Impact Factor
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ABSTRACT: In this study we analyze the usefulness of the histoculture drug response assay (HDRA) based perioperative chemotherapy for non-small cell lung cancer. From 2001 to 2006, we examined the chemosensitivity of 70 lung cancer tissues to cisplatin (CDDP), carboplatin (CBDCA), paclitaxel, docetaxel, gemcitabine and irinotecan. In 16 patients with stage III lung cancer who treated induction therapy, the response rate was 100% of 5 patients treated chemotherapy using 2 HDRA-positive drugs, 50% of 8 patient treeated using 1 positive drugs and 0% of 3 patients treated using negative drugs, respectively. The 3-year survival rate of the 5 patients treated using 2 positive drugs was better than that of 11 patient treated using 1 or non positive drugs (p = 0.07). In 39 patients with stage III lung cancer who treated adjuvant chemotherapy, the survival rate of the 14 patients treated chemotherapy using 2 positive drugs was significantly better than that of 25 patients treated using 1 or non positive drugs (p = 0.03). Therefore, HDRA may useful to the improvement of the response to chemotherapy and survival.
Kyobu geka. The Japanese journal of thoracic surgery 02/2008; 61(1):26-30.
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ABSTRACT: Thymoma is a neoplasm of thymic epithelial cells which is characteristically associated with a large number of non-neoplastic T lymphocytes. These T cells are induced by epithelial cells and show a unique phenotype of CD4(+)8(+) double positive (DP), when studied by flow cytometry. This DP phenotype can be used as one of the diagnostic criteria to indicate a thymoma. The preoperative diagnosis of thymoma and other thymic tumors is an important problem because the treatment differs according to the diagnosis.
A flow-cytometric analysis of needle biopsy specimens was performed on ten thymic tumors. The results were then compared with the findings of a histological diagnosis using conventional hematoxylin-eosin (H&E) staining.
Six cases with high frequencies of DP cells were diagnosed as thymoma by a flow-cytometric analysis, and were confirmed by a histological analysis of the resected specimen. Flow cytometry did not suggest a thymoma in the other four cases with few DP cells. The final diagnoses of the resected specimen of these four cases were: one thymic carcinoma, one malignant lymphoma, and two germ-cell tumors. The accuracy and specificity of diagnosis of thymoma using a fluorescence-activated cell sorting analysis from needle biopsy specimens were 6/6 (100%). On the other hand, the specificity of H&E staining from needle biopsy specimens for the diagnosis of thymoma was 6/6 (100%), but the accuracy was only 6/9 (66.7%).
Flow cytometry can be applied to needle biopsy specimens and thus is considered to offer useful information for the preoperative diagnosis of thymic tumors.
Surgery Today 01/2003; 33(3):163-8. · 1.22 Impact Factor
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ABSTRACT: The Ets-1 protooncogene transcription factor is involved in several cellular functions, including the activation of several proteases, and reportedly plays an important role in carcinoma invasion and metastasis. We hypothesize that Ets-1 mRNA expression could be a predictor of thymoma development and invasion.
Subjects were 37 patients with thymoma whose mRNA expression was quantified by real-time reverse-transcription polymerase chain reaction (RT-PCR) using LightCycler, a microvolume multisample fluorimeter.
No significant difference was found in Ets-1 mRNA expression for gender, age, or pathological subtype. Ets-1 mRNA expression in tumor tissues from stage II-IV thymoma (25.311 +/- 42.336) was more elevated than in that from stage I thymoma (2.097 +/- 4.492) (p = 0.0374). Ets-1 mRNA expression may thus serve as a marker of higher thymoma stages.
With use of the Light Cycler RT-PCR assay, Ets-1 expression may play an important role in the extension and progression of thymoma as in other cancers. Further study and longer follow-up are, however, needed to confirm the Ets-1 impact on biological tumor behavior.
The Japanese Journal of Thoracic and Cardiovascular Surgery 01/2003; 50(12):503-7.
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ABSTRACT: Bcl-2 family proteins regulate programmed cell death, and may play an important role in the selection of lymphocytes. We investigated the expression of Bcl-2, Bcl-x, Bax, Bak and Bim in human lymphocytes using flow-cytometry. Bcl-2 was down-regulated in CD4(+)8(+) (DP) thymocytes and CD19(+)38(+) tonsillar lymphocytes (GC B cells). Among DP thymocytes, cells co-expressing CD69 up-regulated Bcl-2, suggesting that the role of Bcl-2 is promoting survival of positively selected DP cells. Unexpectedly, the expression level of Bcl-x was higher in DP cells than in Single Positive (SP) cells and in CD69(+) DP thymocytes it was lower than in CD69(+) DP thymocytes. Expression of Bim was low in DP thymocytes but high in a subset of GC B cells. Bim and Bax were expressed more highly in SP than in DP thymocytes. Among peripheral blood lymphocytes (PBL), CD8(+) T cells expressed an approximately ten-fold higher level of Bcl-x than CD4(+) T cells while both subsets expressed similar levels of Bcl-2. Bak expression was low and Bim expression was absent in PBL. These results suggest that not only Bcl-2 but other members of the Bcl-2 family are involved in T cell development in the thymus and affinity maturation of B cells in the germinal center.
Immunology Letters 05/2002; 81(2):107-13. · 2.53 Impact Factor
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Hidefumi Sasaki, Masayuki Tanahashi,
Haruhiro Yukiue,
Satoru Moiriyama,
Yoshihiro Kobayashi,
Yoshiaki Nakashima,
Masahiro Kaji,
Masanobu Kiriyama,
Ichiro Fukai,
Yosuke Yamakawa,
Yoshitaka Fujii
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ABSTRACT: Activation of the nuclear hormone receptor perioxisome proliferator-activated receptor gamma (PPARgamma) inhibits cell growth and induces apoptosis in several human cancers. We have hypothesized that PPARgamma mRNA levels could be predictors of the differentiation and survival of lung cancer. The study included 77 lung cancer cases. The mRNA levels were quantified by real time reverse transcription-polymerase chain reaction (RT-PCR) using LightCycler. The PPARgamma mRNA levels were decreased in tumor tissues from lung cancer (0.579 +/- 1.255) compared to the normal adjacent lung tissues (4.191 +/- 2.868) (P = 0.0001). No significant difference in PPARgamma mRNA levels was found among gender, age, and pathological subtype. The PPARgamma mRNA levels were higher in tumor tissues from higher differentiated lung cancer. The NSCLC patients with low PPARgamma mRNA expression (< 0.5) had significantly worse survival than the patients without low PPARgamma mRNA levels (P = 0.0438, Breslow-Gehan-Wilcoxon test; P = 0.0168, Cox's proportional-Hazards regression model). Thus, PPARgamma mRNA levels may serve as a prognostic marker in lung cancer. Using the LightCycler RT-PCR assay, the determination of PPARgamma mRNA levels might provide a potential marker for treatment of lung cancer by PPARgamma agonist. However, further studies and a longer follow up are needed to confirm the impact of PPARgamma in the biological behavior of the tumor.
Lung Cancer 04/2002; 36(1):71-6. · 3.43 Impact Factor
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ABSTRACT: There is an evidence to suggest that cdc25B phosphatase is an oncogenic. We hypothesized that cdc25B gene may be expressed in tumors of patients with non-small cell lung cancer (NSCLC) and affect their clinical outcome. Expression of cdc25B messenger RNA was evaluated by reverse transcription polymerase chain reaction in 55 non-small cell lung carcinomas and adjacent histological normal lung samples using LightCycler. The data was analyzed in reference to clinicopathological data and survival data. There was no difference of cdc25B expression level between the NSCLC tissue and normal lung tissue. There was no relationship between cdc25B gene expression and age, gender, N or T-status and clinical stage. However, the NSCLC patients with high cdc25B expression had significantly poor survival than the patients with low cdc25B expression (P=0.0173). Thus we suggest that cdc25B may predict poor survival.
Cancer Letters.
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悟 森山,
健 山田,
雅幸 棚橋,
雄 彦坂,
泰生 木村,
宏 丹羽,
博 小川,
康司 奥寺,
春彦 椙村,
Satoru Moriyama,
Takeshi Yamada, Masayuki Tanahashi,
Yu Hikosaka,
Hiroshi Niwa,
Haruhiko Sugimura
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ABSTRACT: rights: 日本肺癌学会 rights: 本文データは学協会の許諾に基づきCiNiiから複製したものである relation：isVersionOf: http://ci.nii.ac.jp/naid/110004741270/
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ABSTRACT: Positive selection of immature thymocytes is a developmental process in which TCR ligation with low avidity induces generation of mature T cells. In mouse thymocytes, CD4+8+ double-positive (DP) cells which were treated with a proper combination of calcium ionophore ionomycin and phorbol 12-myristate 13-acetate (PMA) have been reported to differentiate into CD4 single positive cells. However, in human thymocytes the effects of PMA and ionomycin have remained unclear. Here we report that DP cells that were treated with PMA and ionomycin up-regulated bcl-2 and down-regulated CD1 expression. However, CD3 expression remained low. This treatment induced prolonged CD4 down-regulation in DP cells which was an effect also seen in mature peripheral blood T cells. PMA/ionomycin-treated DP cells showed high cell proliferation and resistance to dexamethasone-induced apoptosis. These results indicate that PKC activation and calcium elevation may be part of the biochemical signals that induce positive selection of human DP cells and the system described in this paper may be a useful model to study the signals involved in the selection of human thymocytes.
Human Immunology.
