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Background Bronchogenic cyst sometimes occurs in the mediastinum and rarely in the intrapericardial space. When located in the intrapericardial space, the main vessels or the heart can be compressed. In addition, if it is difficult to deny malignancy or malignancy transformation potential using any modality, surgical resection should be performed. Case presentation The patient was a 21-year-old woman with persistent symptoms similar to a cold. Enhanced computed tomography confirmed a 51 × 36 × 35-mm intrapericardial cystic structure with partial calcification. The lesion was large enough to compress and interfere with the venous return of the superior vena cava. Thrombus formation was suspected upstream of the compression site. We performed utter extirpation through median sternotomy. A histopathological examination of the surgical specimen revealed a bronchogenic cyst. The postoperative course was uneventful, and she was discharged on postoperative day 9. Conclusion We experienced a case of total extirpation of an intrapericardial bronchogenic cyst complicated with compression of the superior vena cava. Long-term follow-up will be necessary.

IntroductionThoracoscopic surgery is performed for refractory or recurrent primary spontaneous pneumothorax (PSP). To reduce postoperative recurrence, additional treatment is occasionally adopted during surgery after bulla resection. However, the most effective method has not been fully elucidated. Furthermore, the preference for additional treatment varies among countries, and its efficacy in preventing recurrence must be evaluated based on settings tailored for the conditions of a specific country. The number of registries collecting detailed data about PSP surgery is limited. Therefore, to address this issue, a prospective multicentre observational study was performed.Methods and analysisThis multicentre, prospective, observational study will enrol 450 participants aged between 16 and 40 years who initially underwent PSP surgery. Data about demographic characteristics, disease and family history, surgical details, and CT scan findings will be collected. Follow-up must be conducted until 3 years after surgery or in the event of recurrence, whichever came first. Patients without recurrence will undergo annual follow-up until 3 years after surgery. The primary outcome is the rate of recurrence within 2 years after surgery. A multivariate analysis will be performed to compare the efficacy of different surgical options. Then, adverse outcomes correlated with various treatments and the feasibility of treatment methods will be compared.Ethics and disseminationThis study was approved by the local ethics committee of all participating centres. The findings will be available in 2025, and they can be used as a basis for clinical decision-making regarding appropriate options for the initial PSP surgery.Trial registration numberNCT04758143.

We have experienced a case of delayed tracheal perforation after pulmonary resection using soft coagulation system. A 58-year-old male underwent operation for primary lung cancer. A soft coagulation system was used for oozing near upper mediastinal lymph nodes. The patient was discharged on postoperative day 8 in a good condition, however sudden tracheal perforation and was occurred on postoperative day 30. An emergency operation revealed that improper use of the soft coagulation system might cause a necrosis of the bronchial wall. Although, a soft coagulation system is useful to control bleeding from small vessels such as bronchial arteries and lymph nodes, this system is different from conventional electrocautery and requires some attention when using.

Purpose The aim of this study was to evaluate the technical and clinical efficacy and safety of CT-guided hookwire marking before video-assisted thoracoscopic surgery (VATS) for small pulmonary lesions. Materials and Methods This procedure was performed on 161 lesions in 154 patients (75 men and 79 women; median age, 62 years; age range 23–89 years). Medical records and images were reviewed, and the technical success rate, surgical success rate and complications were evaluated. Technical success was defined as successful hookwire marking at the target site without marker dropping before VATS. Surgical success was defined as negative surgical margins on pathological examination after VATS. Results There were 97 nodules and 64 ground-glass opacities, and their mean size was 9.8 mm (range 2–34). The technical success rate was 97.5% (157/161). In three of the four failed cases, another hookwire marker was placed, and in the remaining case, VATS was performed without a marker. The surgical success rate was 98.1% (158/161). In the three failed cases, the margin was positive, so lung lobectomy was performed in one case, and the other two cases were observed carefully. Complication rates were as follows: pneumothorax, 37.9% (61/161); focal intrapulmonary haemorrhage, 34.8% (58/139); haemoptysis, 0.6% (1/161); haemothorax, 0% (0/161); air embolism, 0.6% (1/161); dissemination, 0% (0/161); and death, 0% (0/161). Conclusion CT-guided hookwire marking appears to be useful for VATS, but the procedure may, although rarely, cause severe complications such as air embolism.

Birt-Hogg-Dube syndrome (BHD) is an inherited disorder caused by genetic mutations in the folliculin (FLCN) gene. Individuals with BHD have multiple pulmonary cysts and are at a high risk for developing renal cell carcinomas (RCCs). Currently, little information is available about whether pulmonary cysts are absolutely benign or if the lungs are at an increased risk for developing neoplasms. Herein, we describe 14 pulmonary neoplastic lesions in 7 patients with BHD. All patients were confirmed to have germline FLCN mutations. Neoplasm histologies included adenocarcinoma in situ (n = 2), minimally invasive adenocarcinoma (n = 1), papillary adenocarcinoma (n = 1), micropapillary adenocarcinoma (n = 1), atypical adenomatous hyperplasia (n = 8), and micronodular pneumocyte hyperplasia (MPH)-like lesion (n = 1). Five of the six adenocarcinoma/MPH-like lesions (83.3%) demonstrated a loss of heterozygosity (LOH) of FLCN. All of these lesions lacked mutant alleles and preserved wild-type alleles. Three invasive adenocarcinomas possessed additional somatic events: 2 had a somatic mutation in the epidermal growth factor receptor gene (EGFR) and another had a somatic mutation in KRAS. Immunohistochemical analysis revealed that most of the lesions were immunostained for phospho-mammalian target of rapamycin (p-mTOR) and phospho-S6. Collective data indicated that pulmonary neoplasms of peripheral adenocarcinomatous lineage in BHD patients frequently exhibit LOH of FLCN with mTOR pathway signaling. Additional driver gene mutations were detected only in invasive cases, suggesting that FLCN LOH may be an underlying abnormality that cooperates with major driver gene mutations in the progression of pulmonary adenocarcinomas in BHD patients.

症例は27歳女性．症状はなく胸部異常影を指摘されて当科を受診した．胸部レントゲン・CTにおいて，境界明瞭・内部均一な径2cmの円形結節を認めた．胸腔鏡下肺部分切除を行い肺胞腺腫の診断を得た．肺胞腺腫（alveolar adenoma）は極めてまれな良性腫瘍である．病理組織学的にはII型肺胞上皮細胞の嚢胞性の増殖を特徴とする．良性腫瘍であるが確定診断には完全切除を要する．画像で良性腫瘍が疑われ生検で診断の得られない症例は，本疾患を考慮すべきであると考えられた．

Secreted frizzled related protein 1 (SFRP 1) is an antagonist of the transmembrane frizzled receptor, a component of the Wnt signaling pathway, and has been suggested to be a candidate tumor suppressor in several human malignancies. Since SFRP 1 is located at chromosome 8 p 11, where lung cancers also exhibit frequent allelic loss, we hypothesized that the inactivation of SFRP 1 is also involved in lung carcinogenesis. To substantiate this, we performed mutational analysis of SFRP 1 for 29 non-small-cell lung cancer (NSCLC) and 25 small-cell lung cancer (SCLC) cell lines, and expression analysis for the same cell lines. Although somatic mutations were not detected in the coding sequence, downregulation of SFRP 1 was observed in 14 (48%) NSCLC and nine (36%) SCLC cell lines. We analysed epigenetic alteration of the SFRP 1 promoter region and detected hypermethylation in 15 (52%) of 29 NSCLC cell lines, two (8%) of 25 SCLC cell lines, and 44 (55%) of 80 primary lung tumors. By comparing the methylation status with SFRP 1 expression, we found a significant correlation between them. We also performed loss of heterozygosity (LOH) analysis and found that 15 (38%) of 40 informative surgical specimens had LOH in the SFRP 1 gene locus. Furthermore, we performed colony formation assay of two NSCLC cell lines (NCI-H 460 and NCI-H 2009) and found the reduction of colony formation with SFRP 1 transfection. In addition, we also detected that SFRP 1 inhibits the transcriptional activity of beta-catenin, which is thought to be a downstream molecule of SFRP 1, with luciferase reporter assay. Our current studies demonstrated that the SFRP 1 gene is frequently downregulated by promoter hypermethylation and suppresses tumor growth activity of lung cancer cells, which suggests that SFRP 1 is a candidate tumor suppressor gene for lung cancer.

症例は66歳, 女性. 2000年より後縦隔に7×5cmの嚢胞性病変の存在が指摘されていた. 2003年6月より発熱, 咳嗽を繰り返し, 同年10月31日に胸部CT上腫瘤内に鏡面形成が認められたため, 嚢胞内感染を伴う縦隔嚢胞の肺穿破を疑い手術を行った. 腫瘤は壁肥厚を認める単房性の嚢胞で, 炎症のため周囲との癒着が強固であった. 食道, 大動脈に接している壁の一部を残して嚢胞を右肺下葉の一部とともに摘出した. 嚢胞壁は炎症細胞の浸潤が著明で, 上皮の確認はできなかった. 病理学的には確定診断には至らないものの, 摘出組織に食物残渣様異物を認めたことと傍食道に存在していたことから, 食道嚢胞に炎症が発生し, それにより嚢胞の肺穿破が引き起こされたと考える.

誤嚥による肺内伏針に対して,胸腔鏡補助下に伏針のみを摘出しえた症例を経験したので文献的考察を加えて報告する.患者は37歳,女性.咳嗽と血痰を主訴に当院を受診し,胸部X線写真と胸部CTで左肺下葉に肺内伏針を認めた.気管支鏡下の摘出を試みたが,伏針はさらに奥に進んだため摘出困難となり胸腔鏡補助下の摘出を試みた.小開胸部からの触診にて左S9bの胸膜直下にまち針の頭を触知し,ハーモニックスカルペルを用いて臓側胸膜を切開した.露出したまち針の頭を把持鉗子にて牽引し摘出しえた.術後経過良好で第10病日に退院となった.

We have found that a malignant mesothelioma cell line, NCI-H28, had a chromosome 3p21.3 homozygous deletion containing the beta-catenin gene (CTNNB1), which suggested that the deletion of beta-catenin might have a growth advantage in the development of this tumor. To determine whether beta-catenin has a growth-inhibitory activity, we transfected wild-type beta-catenin, Ser37Cys mutant beta-catenin as an activated type, and C-terminus deletion mutant beta-catenin that lacks the transcription activity, into the NCI-H28 cells. A non-small cell lung cancer cell line, NCI-H1299, which expressed endogenous beta-catenin, was also studied. We tested the localization of exogenous beta-catenin in the NCI-H28 cells with immunofluorescence, and found that the wild-type beta-catenin and the C-terminus deletion mutant were more strongly expressed in the plasma membrane and cytoplasm than in the nucleus, while the Ser37Cys mutant was more in the nucleus than in the cytoplasm. By using luciferase-reporter assay, the beta-catenin/T-cell factor 4-mediated transactivity of the Ser37Cys mutant was shown to be higher than that of the wild-type beta-catenin in both cell lines. However, the transactivity of the C-terminus deletion mutant was strongly reduced in both. Colony formation of the NCI-H28 cells was reduced by 50% after transfection with the wild-type beta-catenin, and 60% with the Ser37Cys mutant, but only 20% with the C-terminus deletion mutant compared to the vector control. Inhibition of colony formation in NCI-H28 cells was because of apoptosis, manifested by positive staining of Annexin V and TUNEL assays in transfected cells. In contrast, when transfected with the wild-type beta-catenin, no significant reduction in colony formation was seen in beta-catenin wild-type NCI-H1299 cells. In conclusion, our data indicate that inactivation of beta-catenin by a 3p21.3 homozygous deletion might be a crucial event in the development of the mesothelioma NCI-H28 cells. Thus, while beta-catenin is well known to be a positive growth-stimulating factor for many human cancers, it can also act as a potential growth suppressor in some types of human cancer cells.

We established a new lung cancer cell line, designated Y-ML-1B, from a lung cancer of a 70-year-old Japanese man with leukocytosis and thrombocytosis. Before surgical resection, the white blood cell and platelet counts were elevated to 34,400/mm 3 and 668,000/mm 3 , respectively, and the granulocyte colony-stimulating factor (G-CSF) level in the serum was increased at 141 pg/mL. The primary tumor showed an undifferentiated morphology with large cells and induced extensive thickening of the pleura in the right hemithorax. The Y-ML-1B cells grow as a monolayer, with a doubling time of 19 hours, and are tumorigenic in nude mice, which showed a morphology similar to the primary tumor in xenografts. Analysis of the supernatant of cell culture medium of Y-ML-1B showed elevated levels of G-CSF and other cytokines such as interleukin (IL)-6, IL-8, and granulocyte-macrophage colony-stimulating factor (GM-CSF), consistent with the high levels detected in the patient's serum. Cytogenetic analysis revealed aneuploidy of greater than 56 in metaphases with many structural abnormalities. Mutation analysis of the tumor suppressor genes showed that Y-ML-1B is inactivated in TP53 and RASSF1A , but not in p14 ARF , p16 INK4A , or RB . Neither activating mutations of KRAS or NRAS nor amplification of MYC or MDM2 were detected. Y-ML-1B expressed N-cadherin but not E-cadherin. This newly established cell line might serve as a useful model for studying the molecular pathogenesis for large cell cancers of the lung which express high levels of cytokines.

血気胸の長期放置により線維胸を呈した1例を経験した.症例は28歳の男性で, 1998年6月に胸痛, 発熱で近医にて感冒として内服治療を受けた. 2カ月後, 胸部X線写真で左肺の虚脱と胸水貯留を認め, 当院呼吸器科を受診した. 胸膜炎の診断で入院. 胸腔穿刺にて血性胸水を認め, 持続胸腔ドレナージを行ったが虚脱肺の再膨張を得られず, 逆行性感染による膿胸を合併した. 9月18日外科転科となり, 胸腔鏡下洗浄ドレナージを施行した. 約1カ月後, 胸水からの排菌は消失したものの, 発熱は続き膿胸腔が残存したため, 肺剥皮術を行った. 充分な肺の再膨張が得られ, 術後経過は良好で11月20日に退院となった. 血気胸原因の線維胸では容易な肺の剥皮が期待できることから早期の手術を考慮すべきと考えられた.

The aim of this study was to evaluate the technical and clinical efficacy and safety of CT-guided hookwire marking before video-assisted thoracoscopic surgery (VATS) for small pulmonary lesions.This procedure was performed on 161 lesions in 154 patients (75 men and 79 women; median age, 62 years; age range 23-89 years). Medical records and images were reviewed, and the technical success rate, surgical success rate and complications were evaluated. Technical success was defined as successful hookwire marking at the target site without marker dropping before VATS. Surgical success was defined as negative surgical margins on pathological examination after VATS.There were 97 nodules and 64 ground-glass opacities, and their mean size was 9.8 mm (range 2-34). The technical success rate was 97.5% (157/161). In three of the four failed cases, another hookwire marker was placed, and in the remaining case, VATS was performed without a marker. The surgical success rate was 98.1% (158/161). In the three failed cases, the margin was positive, so lung lobectomy was performed in one case, and the other two cases were observed carefully. Complication rates were as follows: pneumothorax, 37.9% (61/161); focal intrapulmonary haemorrhage, 34.8% (58/139); haemoptysis, 0.6% (1/161); haemothorax, 0% (0/161); air embolism, 0.6% (1/161); dissemination, 0% (0/161); and death, 0% (0/161).CT-guided hookwire marking appears to be useful for VATS, but the procedure may, although rarely, cause severe complications such as air embolism.

NH3 and NaOH/NH3 treated cotton fabrics are processed with hot water at 100 and 130°C, and excellent shrink resistance is obtained with the processing. The crystallite form is transformed from cellulose III to cellulose I. Crystallinity decreased by the NH3 treatment increases a little after hot water processing and washing shrinkage is consider ably controlled. Although the dyeing rate decreases with the NH3 treatment, it increases a little with subsequent hot water processing. In contrast, equilibrium dye uptake de creases with the processing. The mechanical properties of the processed fabrics are evaluated by means of KES shear behavior ( G, 2 HG) and bending hysteresis ( B, 2HB). G and 2HG decreased by NH3 and NaOH/NH3 show almost no change after hot water processing, while B and 2HB, especially the latter, decrease considerably with the hot water processing after treatment with NH3 and NaOH/NH 3.

Desized and scoured cotton fabric is treated in a two-step process with sodium hydroxide/ liquid ammonia and liquid ammonia/sodium hydroxide to investigate the effect of the treat ment sequence. Crystallinity, moisture regain, water absorbency, dyeability, and KES mechan ical properties are measured. Despite a considerable decrease in crystallinity with both treatments, water absorbency decreases with NaOH/NH3 and increases with NH3/NaOH compared with the untreated fabric. Furthermore, the NaOH/NH3 treatment decreases the early dyeing rate with CI Direct Blue 1, whereas the NH 3/NaOH treatment increases the rate remarkably compared to the untreated fabric. Nevertheless, saturation dye uptake with the same dye increases with both treatments compared with the untreated. As a measure of fabric hand, shearing and bending hysteresis curves are obtained with a KES instrument. The shearing modulus and shearing hysteresis width of the NaOH/NH3 treated fabric are much smaller than those of the NH3 treated fabric, while those of the NH3/NaOH treated fabric are smaller compared with the NaOH treated fabric. From these results, it is clear that two-step processes such as NaOH/NH3 and NH3/NaOH are much more effective for softening hand than the NH3 and NaOH treatments independently.

Liquid ammonia and cellulase treatments are known to be effective in improving the soft hand of cotton fabrics. In this study, cotton fabric is treated with liquid ammonia, sodium hydroxide/liquid ammonia, and subsequently with cellulase. Shearing and bending hysteresis curves are measured with a KES instrument. Shearing G, bending modulus B, and hysteresis widths 2HG and 2HB are calculated on the basis of the curves. G, B, 2HG, and 2HB decrease with the NH3 treatment, especially with NaOH/NH3. Although inde pendent cellulase treatment decreases the parameters and produces a soft handle, the combination with NH3 , NaOH/NH3 and with susequent cellulase is much more effective at improving hand.

BACKGROUND Despite the many studies of genetic alterations in advanced lung carcinomas, few reports have analyzed early stage adenocarcinoma of the lung. METHODS We focused on small pulmonary adenocarcinomas, classified according to recently proposed histological criteria (Noguchi M., et al. Cancer 1995;75:2844-52) which divided adenocarcinomas 2 cm or less in diameter into two groups; one showing replacing growth of the pulmonary alveolar structure [A, localized bronchioloalveolar carcinoma (LBAC); B, LBAC with alveolar collapse; C, LBAC with active fibroblastic proliferation] and the other showing non-replacing growth (D, poorly differentiated adenocarcinoma). Ninety-four small pulmonary adenocarcinomas, including 40 of type A and B, 30 of type C and 24 of type D, were examined for loss of heterozygosity (LOH) using microsatellite markers. RESULTS The frequencies of LOH were 19.8% in types A and B, 26.8% in type C and 32.7% in type D tumors. There were no significant differences in the frequency of LOH on chromosomes 2p, 3p, 9p and 17q among tumor types. However, on 17p, the frequency of LOH was significantly lower for types A and B than for type C or D. Three out of six type C tumors which were positive for LOH at several loci showed different LOH patterns in two areas (central and peripheral regions). CONCLUSIONS Allelic losses were detected in very early adenocarcinomas and the frequency of LOHs on chromosome 17p increased during malignant progression of the tumor. Heterogeneous genetic alterations were demonstrated even in small pulmonary adenocarcinomas.

Cefaclor (CCL) and ceftibuten (CETB) are cephem antibiotics synthesized in Japan for oral use. CCL has a broad antibacterial spectrum and CETB is specific for gram-negative bacteria. In our studies, serum and lung tissue CCL and CETB levels were determined from samples obtained at the time of lung surgery after administration of a single fasting dose of 500 mg of CCL or 200 mg of CETB. This study was undertaken to investigate the transfer of the drugs into lung tissue and to determine the appropriate dosing schedules of these drugs for patients with respiratory tract infections. The mean peak serum CCL level was 4.04 μg/mL at 1.5 hours after administration, and the mean alveolar tissue CCL level was 0.30 μg/g tissue 5 hours after administration. The mean peak serum CETB level was 8.47 μg/mL 2 hours after administration, and the mean alveolar, subsegmental bronchial, and bronchiolar tissue CETB levels were 2.65,1.64 and 1.75 μg/g tissue, respectively, at 5 hours after administration, while the CETB level in cancerous lung tissue was 1.41 μg/g 5 hours after administration. From our results, both CCL and CETB showed favorable uptake into lung tissue, suggesting that both compounds may be useful for respiratory tract infections. CCL should be selected for patients with respiratory tract infections in which the pathogens are unknown, with a dose of 500 mg administered every 8 hours. When the pathogen has been identified as a gram-negative organism in patients with pulmonary or bronchiolar infections, 200 mg of CETB should be administered every 12 hours.

Both scoured and sodium hydroxide mercerised cotton fibres have been treated with liquid ammonia atndash33.4°C using commercial operating equipment and then the ammonia removed at 130°C in a hot drum. The moisture regain and water absorbency of the fibres treated with liquid ammonia were increased compared with values on untreated fibres, whereas both these parameters on mercerised cotton fibres were decreased by subsequent liquid ammonia treatment. The fibres were dyed with CI Direct Red 2 and CI Direct Blue 1. Liquid ammonia treatment of the scoured cotton fibres increased the rate of dyeing, equilibrium dye adsorption, standard affinity, heat of dyeing and change of entropy, while the dyeing properties of mercerised cotton were adversely affected by subsequent liquid ammonia treatment.

Activating mutations of EGFR are found frequently in a subgroup of patients with non-small cell lung cancer (NSCLC) and are highly correlated with the response to gefitinib and erlotinib. In the present study, we searched for mutations of EGFR, HER2 and KRAS in 264 resected primary NSCLC from Japanese patients and determined whether there is a correlation between genetic alterations of these genes and clinicopathological factors, together with 85 tumors that we reported previously. EGFR mutations were found in 102 of the total 349 tumors, and seven tumors had two missense mutations. Reverse transcription-polymerase chain reaction of EGFR and subsequent subcloning analyses identified that the double mutations occurred in the same allele. Furthermore, in 202 NSCLC analyzed by Southern blotting, we identified 11 tumors with gene amplification of EGFR, with eight tumors containing a mutation in EGFR. Sequence analysis detected only weak or no signals of the wild-type allele in the eight tumors, strongly suggesting that the mutated allele was amplified selectively. These findings indicate that a dual genetic change of EGFR can occur in the same allele either with a possible second-hit mutation or with amplification, which may imply a more selective growth advantage in a cancer cell. Meanwhile, HER2 mutations and amplifications were found in six of 349 tumors and three of 202 tumors, respectively, and KRAS mutations in 21 of 349 tumors. Mutations of the EGFR and HER2 genes were more frequently found in female never or light-smoking patients with adenocarcinoma, and there were no tumors that had two or more mutations simultaneously among EGFR, HER2 and KRAS. The current study further demonstrates that a double genetic event in EGFR can occasionally occur in lung cancer, thus providing new clues for understanding the involvement of epidermal growth factor receptor signaling cascades in the pathogenesis of NSCLC.

A 46-year-old man was referred to our hospital for the treatment of lung cancer. Computed tomography showed a well-defined tumor mass that was 50×45 mm in size and contained a trabecular pattern of calcification. Since he was diagnosed as having a primary lung adenocarcinoma (clinical stage IB), a left upper lobectomy with mediastinal lymph node dissection was performed. Histologically, the tumor was a poorly differentiated adenocarcinoma with rich fibrous stroma, in which there were island-shaped bone formation lesions. An immunohistochemical examination showed the expression of bone morphogenic protein-2 within tumor cells, which induce and stimulate bone formation. This finding may elucidate a possible mechanism of heterotopic bone formation.

The epidermal growth factor receptor (EGFR) gene has recently been reported to be mutated in a subset of non-small cell lung cancers (NSCLC), with the mutations being correlated with the patients' drug sensitivity to gefitinib, an EGFR kinase inhibitor. In this study, we searched for EGFR mutations in patients with lung cancer using primary tumor specimens obtained at initial surgery and examined whether their recurrent tumors showed a response to gefitinib depending on the presence of the activating mutation. Among 12 lung cancers that were treated with gefitinib after recurrence, we found that all four tumors which showed a response to gefitinib had an activating mutation in EGFR, whereas none of the remaining eight tumors had a mutation. Southern blot analysis showed that two of the four responsive tumors had the EGFR gene amplification. We also examined another 73 NSCLC specimens (47 males and 26 females; 53 adenocarcinomas and 20 non-adenocarcinomas) which were not treated with gefitinib to determine whether NSCLCs with an EGFR mutation have different clinicopathological properties and/or unique genetic alterations of the other cancer-associated genes. We found that 13 (18%) of 73 tumors had a mutation of the EGFR gene, with the most being detected in female adenocarcinomas. Comparing the alterations in KRAS and P53 with the EGFR mutation, we found that 10 tumors with the KRAS mutation did not have an EGFR mutation, suggesting that each mutation occurs exclusively during the development of lung cancer. These results suggest that the mutation analysis of the EGFR gene using the specimens obtained at surgery might be useful in selecting the appropriate treatment(s) for recurrent lung cancer patients.

Non-small cell lung cancer frequently shows loss of heterozygosity of the chromosome 3p21.3 region and several genes such as RASSF1A, BLU, and SEMA3B have been identified as candidate tumor suppressor genes at this region since their downregulation and hypermethylation at their promoter regions were frequently detected in lung cancer. To determine whether these three genes are simultaneously inactivated during lung cancer development, we studied 138 primary non-small cell lung cancers for the promoter methylation status of these genes and allelic loss of the chromosome 3p21.3 region. We found promoter hypermethylation at 32% in RASSF1A, 30% in BLU, and 47% in SEMA3B. Allelic loss of 3p21.3 was detected in 54 (58%) of 93 informative tumors. Despite the weak association of methylation status among these three genes, there was no correlation between the methylation status of each gene and loss of heterozygosity. We also studied possible genes downstream of RASSF1A in 16 primary non-small cell lung cancers and found that the expressions of SM22 and SPARC were significantly downregulated in RASSF1A-hypermethylated tumors. Our results showed that, while candidate tumor suppressor genes at this locus can be simultaneously inactivated by epigenetic alterations, loss of heterozygosity without any hypermethylation of the three genes can also occur in some cases, suggesting that just one allelic loss might also be sufficient for the inactivation of any of these genes for lung cancer development.

BACKGROUND Lung carcinomas show frequent inactivation of the p53 tumor suppressor, which regulates an apoptotic pathway. The objective of the current study was to assess how the p53 apoptotic pathway is altered in nonsmall cell lung carcinoma (NSCLC), especially in tumors without p53 alterations. METHODS p53, its upstream regulators (p14ARF and HDM2), and downstream effectors of the apoptotic pathway (BAX and BCL2) were studied in 118 NSCLC specimens. p53 was analyzed by single-stranded conformation polymorphism analysis covering exons 2–11 and by immunohistochemistry (IHC). p14ARF was analyzed by methylation-specific polymerase chain reaction and IHC. HDM2 was analyzed using Southern blot analysis and IHC. BAX and BCL2 were analyzed by IHC. Two other upstream regulators that regulate the stability of HDM2, PTEN and HAUSP, also were studied. RESULTS Of 118 NSCLC specimens that were analyzed, p53 alterations were detected in 74 tumors (63%), p14ARF inactivation was detected in 53 tumors (45%), and overexpression of HDM2 was found in 31 tumors (26%), including 6 tumors with gene amplification. Although p53 inactivation and HDM2 overexpression were detected simultaneously, HDM2 gene amplification was observed only in tumor specimens without p53 mutations. IHC revealed PTEN down-regulation in 22 of 88 tumors (25%). HAUSP Northern blot analysis demonstrated several-fold differences in gene expression that did not correlate with p53 alterations. Of 118 NSCLC specimens, expression of BAX and BCL2 expression were detected in 46 tumors (39%) and 17 tumors (14%), respectively. Finally, ASPP1 and ASPP2, molecules involved in mediating the transcription function of p53, were not found to be aberrantly expressed when tested by Northern blot analysis. CONCLUSIONS Overall, two or more p53 pathway components were found to be frequently altered in patients with NSCLC. Greater than 90% of the alterations were due to abnormalities of p53, p14ARF, or HDM2. Therefore, the inactivation of one or more components of the p53 pathway appears to be a prerequisite for the development of most NSCLCs. Cancer 2004. © 2004 American Cancer Society.

Activating mutations of RAS gene families have been found in a variety of human malignancies, including lung cancer, suggesting their dominant role in tumorigenesis. However, several studies have shown a frequent loss of the wild-type KRAS allele in the tumors of murine models and an inhibition of oncogenic phenotype in tumor cell lines by transfection of wild-type RAS, indicating that wild-type RAS may have oncosuppressive properties. To determine whether loss of wild-type KRAS is involved in the development of human lung cancer, we investigated the mutations of KRAS, NRAS and BRAF in 154 primary non-small cell lung cancers (NSCLCs) as well as 10 NSCLC cell lines that have been shown to have KRAS mutations. We also determined the loss of heterozygosity status of KRAS alleles in these tumors. We detected point mutations of KRAS in 11 (7%) of 154 NSCLCs, with 10 cases at codon 12 and 1 at codon 61, but no mutations of NRAS or BRAF were found. Using the laser capture microdissection technique, we confirmed that 9 of the 11 tumors and 7 of the 10 NSCLC cell lines retained the wild-type KRAS allele. Among the cell lines with heterozygosity of mutant and wild-type KRAS, all of the cell lines tested for expression were shown to express more mutated KRAS than wild-type mRNA, with higher amounts of KRAS protein also being expressed compared to the cell lines with a loss of wild-type KRAS allele. In addition, among 148 specimens available for immunohistochemical analysis, 113 (76%) showed positive staining of KRAS, indicating that the vast majority of NSCLCs continue to express wild-type KRAS. Our findings indicate that the wild-type KRAS allele is occasionally lost in human lung cancer, and that the oncogenic activation of mutant KRAS is more frequently associated with an overexpression of the mutant allele than with a loss of the wild-type allele in human NSCLC development. © 2003 Wiley-Liss, Inc.

The beta-catenin gene (CTNNB1) has been shown to be genetically mutated in various human malignancies. To determine whether the beta-catenin gene is responsible for oncogenesis in thoracic malignancies, we searched for the mutation in 166 lung cancers (90 primary tumors and 76 cell lines), one blastoma and 10 malignant mesotheliomas (two primary tumors and eight cell lines). Among the lung cancers, including 43 small cell lung cancers (SCLCs) and 123 non-small cell lung cancers (NSCLCs), we identified four alterations in exon 3, which is the target region of mutation for stabilizing beta-catenin. One primary adenocarcinoma had a somatic mutation from C to G, leading to an amino acid substitution from Ser to Cys at codon 37. Among the cell lines, SCLC NCI-H1092 had a mutation from A to G, leading to an Asp to Gly substitution at codon 6, NSCLC HCC15 had a mutation from C to T, leading to a Ser to Phe substitution at codon 45, and NSCLC NCI-H358 had a mutation from A to G, leading to a Thr to Ala substitution at codon 75. One blastoma also had a somatic mutation from C to G, leading to a Ser to Cys substitution at codon 37. Among the 10 malignant mesotheliomas, we identified a homozygous deletion in the NCI-H28 cell line. Cloning of the rearranged fragment from NCI-H28 indicated that all the exons except exon 1 of the beta-catenin gene are deleted and that the deletion junction is 13 kb downstream from exon 1. Furthermore, Northern blot analysis of 26 lung cancer and eight mesothelioma cell line RNAs detected ubiquitous expression of the beta-catenin messages except NCI-H28, although Western blot analysis showed that relatively less amounts of protein products were expressed in some of lung cancer cell lines. Our findings suggest that the beta-catenin gene is infrequently mutated in lung cancer and that the NCI-H28 homozygous deletion of the beta-catenin gene might indicate the possibility of a new tumor suppressor gene residing in this region at 3p21.3, where various types of human cancers show frequent allelic loss.

We report a case of a 40-year-old man with chylopericardium who developed purulent pericarditis caused by Salmonella enteritidis. Thoracoscopic pericardiectomy with debridement effectively controlled the pericardial infection.