Induction of ER Stress-Mediated Apoptosis by alpha-Lipoic Acid in A549 Cell Lines

BACKGROUND: alpha-Lipoic acid (alpha-LA) has been studied as an anticancer agent as well as a therapeutic agent for diabetes and obesity. We performed this study to evaluate the anticancer effects and mechanisms of alpha-LA in a lung cancer cell line, A549. MATERIALS AND METHODS: alpha-LA-induced apoptosis of A549 cells was detected by fluorescence-activated cell sorting analysis and a DNA fragmentation assay. Expression of apoptosis-related genes was analyzed by western blot and reverse transcription-polymerase chain reaction analyses. RESULTS: alpha-LA induced apoptosis and DNA fragmentation in A549 cells in a dose- and time-dependent manner. alpha-LA increased caspase activity and the degradation of poly (ADP-ribose) polymerase. It induced expression of endoplasmic reticulum (ER) stress-related genes, such as glucose-regulated protein 78, C/EBP-homologous protein, and the short form of X-box binding protein-1, and decreased expression of the anti-apoptotic protein, X-linked inhibitor of apoptosis protein. Reactive oxygen species (ROS) production was induced by alpha-LA, and the antioxidant N-acetyl-L-cysteine decreased the alpha-LA-induced increase in expression of apoptosis and ER stress-related proteins. CONCLUSION: alpha-LA induced ER stress-mediated apoptosis in A549 cells via ROS. alpha-LA may therefore be clinically useful for treating lung cancer.

The Ability of FDG Uptake Ratio and Glut-1 Expression to Predict Mediastinal Lymph Node Metastasis in Resected Non-small Cell Lung Cancer / 대한흉부외과학회지

BACKGROUND: This study was designed to evaluate the FDG uptake ratio of mediastinal node and primary tumors using integrated PET/CT imaging combined with Glut-1 expression of the primary tumor in order to predict the N2 status more accurately in NSCLC patients. MATERIAL AND METHOD: Patients who underwent integrated PET/CT scanning with a detectable mSUV for both primary tumors and mediastinal lymph nodes were eligible for this study. The FDG uptake ratio between the mediastinal node and the primary tumor was calculated. RESULT: The average mSUV of primary tumors and mediastinal nodes were, respectively, 7.4+/-2.2 and 4.2+/-2.2 in N2-positive patients and 7.6+/-3.7 and 2.8+/-6.9 in N2-negative patients. The mean FDG uptake ratio of mediastinal node to primary tumor were 0.58+/-0.23 for malignant N2 lymph nodes and 0.45+/-0.20 for benign lymph nodes (p<0.05). Models which combined Glut-1 expression with an FDG ratio have better diagnostic power than models that use the FDG uptake ratio alone. CONCLUSION: In some patients with a previous history of pulmonary tuberculosis or other inflammatory lung diseases, an FDG uptake ratio combined with Glut-1 expression may be useful in diagnosing mediastinal node metastasis more exactly.

Tumorigenesis after Injection of Lung Cancer Cell Line (SW-900 G IV) into the Pleural Cavity of Nude Mice / 대한흉부외과학회지

BACKGROUND: Base on types of tumor, the types of expressed tumor is diverse and the difference in its expression rate is even more various. Due to such reasons an animal model is absolutely needed for a clinical research of lung cancer. The author attempted oncogenesis by cultivating a cell line of non-small cell carcinoma and then injecting it inside thoracic cavities of nude mice. The author conducted quantitative analyses of HER2/neu tumor gene - an epidermal growth factor receptor (EGFR) related to lung cancer, and TGF-beta1, which acts as a resistance to cell growth inhibition and malignant degeneration. In order to investigate achievability of the oncogenesis, histological changes and the expression of cancer gene in case of orthotopic lung cancer is necessary. MATERIAL AND METHOD: Among 20 immunity-free male BALB/c, five nude mice were selected as the control group and rest as the experimental group. Their weights ranged from 20 to 25 gm (Orient, Japan). After injection of lung cancer line (SW900 G IV) into the pleural cavity of nude mice, They were raised at aseptic room for 8 weeks. HER2/neu was quantitatively analyzed by separating serum from gathered blood via chemiluminiscent immunoassay (CLIA), and immunosandwitch method was applied to quantitatively analyze TGF-beta1 SPSS statistical program (SPSS Version 10.0, USA) was implemented for statistical analysis. Student T test was done, and cases in which p-value is less than 0.05 were considered significant. RESULT: Even after lung cancer was formed in the normal control group or after intentionally injected lung cancer cell line, no amplification of HER2/neu gene showed reaction. However, the exact quantity of TGF-beta1was 28,490+/-8,549 pg/mL, and the quantity in the group injected with lung cancer cell was 42,362+/-14,449 pg/mL, meaning 1.48 times highly significant (p<0.483). It proved that HER2/neu gene TGF-beta1had no meaningful interconnection. CONCLUSION: TGF-beta1gene expressed approximately 1.48 times amplification in comparison to the control group. The amplification of TGF-beta1meant somatic recuperation inhibition mechanism due to carcinogenesis in nude mice was definitely working. It may be implemented as a quantitative analysis that allows early detection of lung cancer in human body.

The Caspase-3 and c-myc Expressions in Completely Resected Non-small Cell Lung Cancer and Its Prognostic Significance / 대한흉부외과학회지

BACKGROUND: Caspase-3 is a cysteine protease that plays a major role in the process of apoptotic cell death. The dysregulated expression of c-myc contributes to the tumorigenesis in a variety of human cancers. The aim of this study was to investigate the expressions of caspase-3 and c-myc and their significances as prognosis markers in patients with completely resected non-small cell lung cancer (NSCLC). MATERIAL AND METHOD: A total 130 consecutive patients who had undergone complete resection without pre-operative radio-therapy or chemotherapy between May 1996 and December 2003 for NSCLC were retrospectively reviewed. The median follow-up period of the patients was 50 months (range: 3~128 months). The expressions of caspase-3 and c-myc were immunohistochemically examined, and these were correlated with the clinico-pathologic data. RESULT: The prevalence of caspase-3 and c-myc expressions in the patients was 68% (88/130) and 59% (77/130), respectively. Significant association was found between the frequency of the expressions of caspase-3 and c-myc (p=0.025). The caspase-3 and c-myc expressions were not significantly associated with the prognosis in all the patients. However, according to stages, a positive caspase-3 expression was significantly correlated with a favorable prognosis for patients with stage IIIa disease (median survival period: 35 months vs. 10 months, p=0.021). Multivariate analysis showed the pathologic stage to be significantly correlated with a good prognosis in all the patients (p=0.024), and with a positive caspase-3 expression, well differentiated tumor and negative neuronal invasion in the patients with stage IIIa disease (p=0.005, p=0.003, p=0.004, respectively). CONCLUSION: Caspase-3 and c-myc were frequently expressed in NSCLC, suggesting its possible involvement in tumor development. The caspase-3 expression, as determined with performing immunohistochemical staining, may be a favorable prognostic indicator in patients with completely resected NSCLC of an advanced stage (IIIa).

Independent Prognostic Value of the Fascin Expression in Patients with Esophageal Cancer / 대한흉부외과학회지

Backgrond: Fascin is an actin-bundling protein that induces membrane protrusions and it increases cell motility in various transformed cells. Esophageal cancer is one of the most lethal malignancies, and it exhibits extensive local invasion or frequent regional lymph node metastasis even after curative surgery. We investigate the expression of fascin by performing immunohistochemistry to evaluate the clinical characteristics and prognostic significance of its expression in esophageal cancer patients. MATERIAL AND METHOD: Immunochemistry for fascin was performed on 76 tumor samples from 76 patients who underwent esophageal cancer operations. The expression levels of fascin in the 76 esophageal cancer tissues were compared with those in the corresponding normal esophageal epithelium. The fascin-positive samples were defined as those showing more than 75% of fascin-positive cells. RESULT: Overall, a fascin positive expression was detected in 39 (51.3%) out of the total 76 cases. The tumors with positive fascin expression tended to more frequently show a higher stage (p=0.030), and a higher T-factor (p=0.031). The prognosis of the fascin negative group was significantly better than that of the fascin positive group (p=0.004). Multivariate analysis revealed that lymphovascular invasion and the fascin expression were independent prognostic factors. CONCLUSION: Fascin was expressed in 51.3% of the esophageal cancer tissues, and a positive expression of fascin was associated with more advanced tumor progression and recurrence. Our study suggests that the fascin expression may be an independent prognostic factor for an unfavorable clinical course for those patients suffering with esophageal cancer.

Expression of Hypoxia-inducible Factor-1alpha in Non-small Cell Lung Cancer: Relationship to Prognosis and Tumor Biomarkers / 대한흉부외과학회지

BACKGROUND: Tissue hypoxia is characteristic of many human malignant neoplasm, and hypoxia inducible factor-1 (HIF-1) plays a pivotal role in essential adaptive response to hypoxia, and activates a signal pathway for the expression of the hypoxia-regulated genes, resulting in increasing O2 delivery or facilitating metabolic adaptation to hypoxia. Increased level of HIF-1alpha has been reported in many human malignancies, but in non-small cell lung carcinoma the influence of HIF-1alpha on tumor biology, including neovascularization, is not still defined. In present study the relationship of HIF-1alpha expression on angiogenetic factors, relationship between the tumor proliferation and HIF-1alpha expression, interaction of HIF-1alpha expression and p53, and relationship between HIF-1alpha expression and clinico-pathological prognostic parameters were investigated. MATERIAL AND METHOD: Archival tissue blocks recruited in this study were retrieved from fifty-nine patients with primary non-small cell lung carcinoma, who underwent pneumonectomy or lobectomy from 1997 to 1999. HIF-1alpha, VEGF (vascular endothelial growth factor), and p53 protein expression and Ki-67 labeling index in tumor tissues were evaluated, using a standard avidin-biotin-peroxidase complex (ABC) immunohistochemistry. Relationship between the HIF-1alpha expression and VEGF, p53 overexpression and correlation between the HIF-1alpha expresseion and Ki-67 index were analyzed. Clinico-pathologic prognostic parameters were also analyzed. RESULT: HIF-1alpha expression in cancer cells was found in 24 of 59 cases of non-small cell lung carcinoma (40.7%). High HIF-1alpha expression was significantly associated with several pathological parameters, such as pathological TMN stage (p=0.004), pT stage (p=0.020), pN stage (p=0.029), and lymphovascular invasion (p=0.019). High HIF-1alpha expression was also significantly associated with VEGF immunoreactivity (p<0.001), and aberrant p53 expression (p=0.040). but was marginally associated with Ki-67 labeling index (p=0.092). The overall 5-year survival rate was 42.3%. The survival curve of patients with a high HIF-1alpha expression was worse than that of patients with low-expression (p=0.002). High HIF-1alpha expression was independent unfavorable factors with a marginal significance in multivariate analysis performed by Cox regression. CONCLUSION: It is suggested that high HIF-1alpha expression may be associated with intratumoral neovascularization possibly through HIF-VEGF pathway, and high HIF-1alpha expression could be associated with lymph node metastasis and post operative poor prognosis in patients with non-small cell lung ca

A Study on the Expression of CD44s and CD44v6 in Non-Small Cell Lung Carcinomas / 대한흉부외과학회지

BACKGROUND: CD44 is a glycoprotein on the cell surface which is involved in the cell-to-cell and cell-to-matrix interaction. The standard form, CD44s and multiple isoforms are determined by alternative splicing of 10 exons. Recent studies have suggested that CD44 may help invasion and metastasis of various epithelial tumors as well as activation of lymphocytes and monocytes. The expression pattern of CD44 can be different according to tumor types. The author studied the expression pattern of CD44s and one of its variants, CD44v6 in non-small cell lung carcinomas (NSCLC) to find their implications on clinicopathologic aspects, including the survival of the patients. MATERIAL AND METHOD: A total of 89 primary NSCLSs (48 squamous cell carcinomas, 33 adenocarcinomas, and 8 undifferentiated large cell carcinomas) were retrieved during the years between 1985 to 1994. The immunohistochemistry was done by using monoclonal antibodies and the CD44 expression for angiogenesis was evaluated by counting the number of tumor microvessels. RESULT: Seventy-one (79.8%) and 64 (71.9%) among 89 NSCLSs revealed the expression of CD44s and CD44v6, respectively. The expression of CD44s was well correlated with that of CD44v6 (r=0.710, p<0.0001). The expression of CD44s and CD44v6 was associated with the histopathologic type of the NSCLCs, and squamous cell carcinoma was the type that showed the highest expression of CD44s and CD44v6 (p<0.0001). Microvessel count was the highest in adenocarcinomas (113.6+/-69.7 on 200-fold magnification and 54.8+/-41.1 on 400-fold magnification) and correlated with the tumor size of TNM system (r=0.217, p=0.043) and CD44s expression (r=0.218, p=0.040). In adenocarcinoma, the patients with higher CD44s expression survived shorter than those with lower CD44s expression (p=0.0194) but there was no statistical significance on multivariate analysis(p=0.3298). CONCLUSION: The expression of both CD44s and CD44v6 may be associated with the squamous differentiation in non-small cell lung carcinomas. The relationship of CD44s expression with microvessel density of the tumor suggests an involvement of CD44s in tumor angiogenesis, which in turn would help tumor growth.

Clinical Significance of Increased Ki-67 Protein Expression in Non-small Cell Lung Cancers / 대한흉부외과학회지

BACKGROUND: The Ki-67 protein is a biomarker associated with cell proliferation and a valuable negative prognostic factor in non-small cell lung cancer. We investigated the Ki-67 protein expression in resected non-small cell lung cancer to evaluate the impact on clinicopathological characteristics and postoperative prognosis. MATERIAL AND METHOD: Using monoclonal antibody Ki-67, we immunohistochemically examined 38 surgically resected non-small cell lung cancers to determine Ki-67 Labeling Index (LI). We analysed the differences of clinicopathological characteristics and postoperative recurrence and survival between High Ki-67 Group (LI> or =20%) and Low Ki-67 Group (LI<20%). RESULT: The Ki-67 LIs were heterogenous and a mean values was 20.0+/-20.05%. There were no significant differences in age, sex, smoking, TNM stage, and vascular invasion between High Ki-67 Group and Low Ki-67 Group. A High Ki-67 Group was significantly associated with squamous cell type, poor differentiation, and lymphatic invasion (p< or =0.05). High Ki-67 Group showed a trend of lower survival (median 47.2 vs. 96.5 months, p=0.312) and lower disease-free survival (median 18.2 vs. 72.3 months, p=0.327) than Low Ki-67 Group. CONCLUSION: These results indicate that increased Ki-67 protein expression may be a negative prognostic factor and showed a trend of shortened survival and disease-free survival. To evaluate the pivotal role of Ki-67 protein expression, a long-term follow-up and further study are required.

Cytotoxicity of COX-2 Inhibitor (Nimesulide) in Non-small Cell Lung Cancer Cell Line / 대한흉부외과학회지

BACKGROUND: In recent years, a combination of two demographic phenomena, an increased number of older people in the population and an increase in the incidence of lung cancer with age, has made it mandatory to develop therapeutic modalities with less toxicity for the treatment of inoperable elderly patients with lung cancer. Therefore, we investigated the correlation between COX-2 expression and cytotoxicity of Nimesulide, a specific COX-2 inhibitor. MATERIAL AND METHOD: Immunohistochemical staining of COX-2 was performed. After exposure of Nimesulide, XTT analysis, FACS analysis and Hoechst staining were carried out. RESULT: COX-2 protein was expressed in non- treated A549 cells strongly, but not in H1299. Cytotoxicity of Nimesulide against A549 cell and H1299 cell were similar and IC50 of Nimesulide in both cell lines were 70.9 microM in A549 cell line and 56.5 microM in H1299 cell line respectively. FACS analysis showed G0/G1 arrest in both cell lines and the S phase cell fraction was decreased. Morphologic assessment of apoptosis by Hoechst 33258 staining, many apoptotic cells were detected in both cell lines. CONCLUSION: Selective COX-2 inhibitor, Nimesulide, can inhibit the proliferation of non-small cell lung cancer cell lines in vitro. Inhibitory effect of Nimesulide are induction of apoptosis and G0/G1 arrest. There is no correlation between COX-2 expression and cytotoxicity of Nimesulide, a specific COX-2 inhibitor. Therefore, highly selective COX-2 inhibitors such as Nimesulide can be expected to lead to even greater efficacy of their use as adjuncts to various anticancer angents and radiation therapy for the treatment of high-risk patients.

Expression of Hypoxia-inducible Factor-1alpha in Esophageal Squamous Cell Carcinoma: Relationship to Prognosis and Tumor Biomarkers / 대한흉부외과학회지

BACKGROUND: Tissue hypoxia is a characteristic of many human malignant neoplasms, and hypoxia inducible factor-1 (HIF-1) plays a pivotal role in essential adaptive response to hypoxia, and activates a signal pathway for the expression of the hypoxia-regulated genes, resulting in increased oxygen delivery or facilitating metabolic adaptation to hypoxia. Increased level of HIF-1alpha has been reported in many human malignancies, but in esophageal squamous cell carcinoma, the influence of HIF-1alpha on tumor biology, including neovascularization, is not still defined. MATERIAL AND METHOD: The influence of HIF-1alpha expression on angiogenic factors, correlation between the tumor proliferation and HIF-1alpha expression, interaction of HIF-1alpha expression and p53, and correlation between HIF-1alpha expression and clinicopathological prognostic parameters were investigated, using immunohistochemical stains for HIF-1alpha, VEGF, CD34, p53, and Ki-67 on 77 cases of resected esophageal squamous cell carcinoma. RESULT: HIF-1alpha expression in cancer cells was found in 33 of 77 esophageal squamous cell carcinoma cases. The 33 cases (42.9%) showed positive stain for HIF-1alpha. High HIF-1alpha expression was significantly associated with several pathological parameters, such as histologic grade (p=0.032), pathological TMN stage (p=0.002), the depth of tumor invasion (p=0.022), regional lymph node metastasis (p=0.002), distant metastasis (p=0.049), and lymphatic invasion (p=0.004). High HIF-1alpha expression had significant VEGF immunoreactivity (p=0.008) and Ki-67 labeling index (p<0.001), but was not correlated with microvascular density within tumors (p=0.088). The high HIF-1alpha expression was correlated with aberrant p53 accumulation with a marginal significance (p=0.056). The overall 5-year survival rate was 34.9%. The survival rate of patients with a high HIF-1alpha expression was worse than that of patients with low-expression tumors (log-rank test, p=0.0001). High HIF-1alpha expression was independent unfavorable factors although statistical significance is marginal in multivariate analysis. CONCLUSION: It is suggested that (1) high HIF-1alpha expression in esophageal squamous cell carcinoma is associated with tumor hypoxia, or with genetic alteration in early carcinogenesis and progressive stages, (2) high HIF-1alpha expression may be associated with intratumoral neovascularization through HIF-VEGF pathway, and (3) high HIF-1alpha expression is associated with poor prognosis in patients with esophageal squamous cell carcinoma and may play a role as biomarker for regional lymph node metastasis.

Prognostic Significance of Cyclin B1 and p53 Expression in Patient with Esophageal Squamous Cell Carcinoma / 대한흉부외과학회지

BACKGROUND: It has been reported that p53 regulates the G2-M checkpoint transition through cyclin B1, and it has been suggested that p53 plays an important role in the development and progression of various malignancies. The aim of this study is to clarify the role of the cell cycle regulators, cyclin B1 and p53 in patients with esophageal squamous cell carcinoma (ESCC). MATERIAL AND METHOD: Tissue samples from 46 patients with ESCC were included in this study. Expression levels of cyclin B1 and p53 in samples of normal squamous epithelium, dysplasia, and tumor cells from patients with ESCC were analyzed by immunohistochemical study. RESULT: Several cells in the basement layer of normal epithelium expressed cyclin B1. The number of cyclin B1 positive cells tended to increase as the degree of dysplasia increased from low grade to high grade. More than 10% of tumor cells were cyclin B1 positive in 19 patients (41.3%). Several clinicopathologic parameters, including tumor stage (p<0.05), pathologic lymph node status (p<0.05) and invasion of lymphatic vessels (p<0.05), were correlated with the overexpression of cyclin B1. Elevated expression levels of cyclin B1 also correlated with a poor prognosis in patient with ESCC in univariate analysis (p<0.05) and multivariate analysis (p<0.05). In contrast, p53 expression exhibited significant correlation with the level of cyclin B1 expression, but was not associated with prognostic parameters in patients with ESCC. CONCLUSION: These findings suggest that cyclin B1 is involved in the pathogenesis of carcinoma of the esophagus and that elevated levels of cyclin B1 expression, but not p53 expression, may indicate a poor prognosis for patients with ESCC.

Human Lung Cancer Cell Xenografts Implanted under the Capsule of Kidney, Spleen and Liver / 대한흉부외과학회지

BACKGROUND: Complete resection by the surgery has been selected as the treatment of choice in lung cancer patients, but in cases of recurrence after excision or inoperable cases, the importance of anticancer chemotherapy has been emphasized. If one can select a set of the sensitive chemotherapeutic agents before anticancer chemotherapy, it will give more favourable results. Subrenal capsular assay has been recognized as a useful in-vivo chemosensitivity test of thoracic and abdominal tumors and it can be done in a short time for a rapid interpretation of tumor responsiveness to anticancer chemotherapeutic drugs. It has been reported that various kinds of cancer cells can be implantable to the kidney, but so far there is no comparative study of xenogeneic cell implantation on liver, spleen and kidney. The author implanted the human lung cancer cells under the capsule of S.D rat's liver, spleen and kidney respectively and compared the pattern of growth and histology. MATERIAL AND METHOD: After incubation of human lung cancer cell line (SW-900 G IV) in RPMI 1640 (Leibovitz L-15 medium) culture media, 3x3x3 mm size fibrin clots which contain 108 cancer cells were made. Thereafter the fibrin clots were implanted at subcapsule area of liver, spleen and kidney of S.D. female rat. For immune suppression, cyclosporin-A (80 mg/Kg) was injected subcutaneously daily from post-implantation first day to sixth day. The body weight was measured at pre and post implantation periods. The growth pattern and the size of tumor mass were observed and the pathologic examination and serum tumor marker tests were performed. RESULT: Body weight increased in both of control and experimental groups. Serum Cyfra 21-1 was not detected. Serum levels of CEA and NSE revealed no significant change. The SCC-Ag increased significantly in implanted group. The growth rate of human lung cancer cells which was implanted on spleen was higher than on liver or kidney. The surface area, thickness, and volume of tumor mass were predominant at spleen. The success rates of implantation were 80% on kidney, 76.7% on spleen and 43.3% on liver. Pathologic examination of implanted tumors showed characteristic findings according to different organs. Tumors that were implanted on kidney grew in a round shape, small and regular pattern. In the spleen, tumors grew well and microscopic neovascularization and tumor thrombi were also found, but the growth pattern was irregular representing frequent daughter mass. Human lung cancer cells that were implanted in the liver, invaded to the liver parenchyme, and had low success rate of implantation. Microscopically, coagulation necrosis and myxoid fibrous lesion were observed. CONCLUSION: The success rate of implantation was highest in the kidney. And the mass revealed regular growth that could be measured easily. The SCC-Ag was presented earlier than CEA or Cyfra21-1. The Cyfra21-1 was not detected at early time after implantation. The best model for tumor implantation experiment for chemosensitivity test was subrenal capsular analysis than liver and spleen and the useful serum tumor marker in early period of implantation was the SCC-Ag.

Prognostic Significance of Cyclin E and p27 in Stage I Non-Small Cell Lung Cancer / 대한흉부외과학회지

BACKGROUND: Cyclin E plays a pivotal role in the regulation of G1-S transition and could consequently be a deregulated molecule in tumors. The activity of the cdk2-cyclin E complex is increased by degradation of cdk inhibitor p27kip1. Little is known about the expression and prognostic significance of cyclin E and p27 in non-small cell lung cancer(NSCLC). MATERIAL AND METHOD: The expression of cyclin E and p27 in eighty-one cases of resected stage I NSCLC tissues and its relation to major clinico-pathological factors, including histology, differentiation, size of tumor, pleural invasion and survival rate were studied and analyzed. Immunohistochemical analysis with monoclonal antibodies specific for cyclin E and p27 were performed by ABC method. RESULT: Expression rates of cyclin E and p27 in stage I NSCLC tissues were 29.6% and 28.4% respectively. Cyclin E was expressed higher in cases of pleural invasion(p=0.04), and p27 was expressed higher in diameter of tumor less than 3cm(p=0.015). The 5-years survival rate was lower in cases of positive expression of cyclin E than in cases of negative expression of cyclin E(44.4% vs 68.2%, p=0.015), and the 5-years survival rate was 72.2% in positive expression of p27 and 56.2% in negative expression of p27(p=0.09). The 5-years survival rate was higher in negative expression of cyclin E and positive expression of p27 than in cases of positive expression of cyclin E and negative expression of p27 (73.5% vs 36.3%, p=0.0029). In multivariate analysis, expression of cyclin E was an unfavorable prognostic factor(RR=3.578, p=0.006) and p27 was a favorable prognostic factor(RR=0.183, p=0.019) independently. CONCLUSION: Cyclin E and p27 may play a pivotal role for the biological behavior of stage I NSCLC, so that the expressions of cyclin E and p27 may be new prognostic markers.

Expression of Mutant p53 and MAGE-3 Gene Products in Esophageal Squamous Cell Carcinoma / 대한흉부외과학회지

BACKGROUND: Despite recent advances in multimodality therapy, the prognosis for invasive esophageal cancer is poor, with five years survival rate generally below 10%. Therefore, immunotherapy is considered as one of the new therapeutic modality in esophageal cancer. However, expression of tumor specific antigen in tumor tissue should be necessary for immunotherapy of tumor. This study is to clarify that mutant p53 protein and MAGE-3 gene product is expressed in esophageal cancer specifically and they can be played a role of prognostic factors in esophageal cancer. MATERIAL AND METHOD: Expression of mutant p53 protein and MAGE-3 gene products in formalin fixed, paraffin embedded samples of 79 patients with primary squamous cell carcinoma of the esophagus, who undewent esophageal resection, were analyzed immunohistochemically with DO-7 monoclonal antibody and anti- MAGE-3 antibody. Twenty cases of esophageal normal mucosa and 20 cases of leiomyoma which is a benign tumor of esophagus, were used as control groups. Immunoreactivities of mutant p53 and MAGE-3 gene product in esophageal cancer tissues were analyzed and the relationships between immunoreactivity of mutant p53 protein, MAGE-3 gene product and AJCC stage of esophageal cancer were determined by the Chi-square test. RESULT: Positive immunoreactivity of mutant p53 and MAGE-3 gene product were each of 41/79(51.9%), 48/79(60.8%) in esophageal cancer tissue, but 0% in normal mucosa and leiomyoma of esophagus(p<0.001). Both immunoreactivity of mutant p53 and MAGE-3 gene products were not related to AJCC stage of esophageal cancer(p=0.193, p=0.452). There was not correlation between expression of mutant p53 protein and MAGE-3 gene product in esophageal cancer(p=0.697). CONCLUSION: Mutant p53 and MAGE-gene product cannot be a prognostic factor in squamous cell carcinoma of esophagus, but mutant p53 and MAGE-3 gene product is expressed in squamous cell carcinoma of the esophagus specifically, so esophageal cancer can be target for cytotoxic T lymphocyte in anticancer immunotherapy.

A Study of SCC Antigen and EGFr in Tissues of Squamous Cell Carcinoma of Lung / 대한흉부외과학회지

The aim of this study was to evaluate a usefulness of serum SCC antigen in diagnosis or evaluation of therapeutic effect of lung cancer by investigation of the differences of SCC antigen concentration in lung mass according to TNM staging, and mass size of lung cancer. And the other aim was to know whether SCC antigen plays a role in infiltrative growth of lung cancer or not, comparing with concentration of epidermal growth factor receptor (EGFr) in tissue which is related with growth and differentiation of tumor cell. The results of this study were as follows. The concentration of SCC antigen in squamous cell carcinoma of lung (69+/-25ng/ml) was higher than in unaffected lung tissue (34+/-7ng /ml). (p or =5cm in diameter), the concentration of SCC antigen in peripheral portion of tumor was higher than that of central portion. (p<0.05). The concentration of EGFr according to tumor size was not significantly different in central and peripheral portion of tumor. The concentration of SCC antigen according to TNM staging of lung cancer was that from central portion was higher in stage I, II, but that from peripheral portion was higher in stage III, IV (p<0.05). The concentration of EGFr from central portion was higher in higher TNM stage (not significant) but that from peripheral portion shows no significant changes. In conclusion, the concentration of SCC antigen in tissue was higher in squamous cell carcinoma than in unaffected lung tissue or adenocarcinoma, and the concentration of SCC antigen increased according to tumor size or TNM staging like in serum level. so, serum SCC antigen is a useful tumor marker to diagnose or evaluate therapeutic effect of squamous cell carcinoma of lung. But further studies are necessary to confirm the relation of infiltrative growth in lung cancer and concentration of SCC antigen because there was a different pattern of regional tissue concentration of SCC antigen and EGFr.

Correlation between Cyclin D1 , p53 , Bcl-2 Protein Expression and Prognosis in Primary , Resected Stage IIIA Non-Small Cell Lung Cancer (NSCLC) / 대한흉부외과학회지

BACKGROUND: Lung cancer formation is a multistage process involving activation of protooncogene and inactivation of tumor suppressor genes. We evaluate the significance of cyclin D1, p53, bcl-2 gene mutations in patients with curatively resected stage IIIA non-small cell lung cancer (NSCLC). MATERIAL AND METHOD: One hundred consecutive cases of stage IIIA lung cancers from patients operated on curatvely between 1990 and 1995 for which adequate paraffin blocks and clinical history were available. Immunohistochemical studies were performed on the representative tissue sections from each case by the labelled streptovidin-biotin method. Sections for cyclin D1, p53, Bcl-2 immunostaining were pretreated in a microwave oven for 10 to 20 minutes in citrate buffer before immunostaining. The overnight incubation with NCL-cyclin D1-GM for cyclin D1, with clone DO-7 for p53, with clone 124 for bcl-2 was done. Mean follow-up was 24.1 months (range 2-84 months) after operation. RESULT: One hundred cases of lung cancers were composed of 56 cases of squamous cell carcinoma, 37 cases of adenocarcinoma, 5 cases of adenosquamous cell carcinoma, and 2 cases of large cell carcinoma. The 5-year survival was 32.1%. The positive expression rate of cyclin D1 was 35%, p53 was 56%, and bcl-2 was 17%. But there were no correlation between cyclin D1, p53, Bcl-2 protein expression and survival. CONCLUSION: These observation indicate that cyclin D1, p53, bcl-2 protein overexpression might be implicated in the oncogenesis of non-small cell lung carcinomas but they have no usefulness as a prognostic marker.

A Study of the Prognostic Factors in Resected Stage I Non-Small Cell Lung Cancer / 대한흉부외과학회지

BACKGROUND: About 30% to 40% of the patients with pathologic stage I non-small cell lung cancer (NSCLC) die within 5 years after complete resection. The identification of poor prognostic factors and the application of additional treatment are very important to improve the survival rate in resected stage I NSCLC. MATERIALS AND METHODS: Sixty-eight (68) patients who had been diagnosed postoperatively between Janury 1989 and December 1995 as having stage I non-small cell lung cancer according to the TNM classification were studied. The postoperative 5-year survival rate was calculated with the Kaplan-Meier method, and clinico-histopathologic factors including age, sex, operative method, type of tumor cell, T factor, grade of the differentiation in a squamous cell carcinoma, invasion of blood vessel and expression of the nm23-H1 protein were investigated and analyzed. RESULTS: The median survival of the entire group of patients was 58+/-3 months, with a 5-year survival of 58.9%. In univariate analysis, invasion of blood vessel and poor differentiation of the tumor cell in a squamous cell carcinoma significantly worsened the survival. In multivariate analysis, invasion of blood vessel and grade of the differentiation of the tumor cells in a squamous cell carcinoma remained independent prognostic factors. High expression of the nm23-H1 protein was related to a high postoperative 5-year survival in comparision with low expression of the nm23-H1 pretein (73.0% vs 50.7%), but there was no statistical significance. CONCLUSIONS: These results highlight the negative prognostic value of poor differentiation of tumor cells in a squamous cell carcinoma and invasion of blood vessel in stage I non-small cell lung cancer. Also, further studies are necessary to be determined prognostic value of the T factor and expression of the nm23 protein in non-small cell lung cancer.

The Study of CYFRA 21-1 and Epidermal Growth Factor Receptor Levels in Cancer Tissue of Bronchogenic Carcinoma Patients / 대한흉부외과학회지

CYFRA 21-1 is known to be a cytokeratin 19 fragment, and it can be detected by using two specific monoclonal antibodies (KS 19-1 and BM 19-21) and can be clinically applied as a useful circulating tumor marker. The epidermal growth factor receptor (EGF-R) expression was evaluated and characterized by its tyrosine protein kinase activity and by its ligand-stimulated autophosphorylation, a property shared with other peptide growth factor receptors. Autocrine or paracrine action was initiated by a growth factor, or by a transforming growth factor alpha, which had an extensive homology with EGF and which also stimulated tyrosine kinase activity on the EGF-R. The CYFRA 21-1 and the EGF-R levels in 30 patients with primary lung tumors were investigated. There were 24 patients with squamous cell carcinomas and 6 patients with adenocarcinomas. Specimen 5 mm3 in size were sampled at three different locations ; the main lesion, the boundary between the lesion and the unaffected tissue, and the unaffected tissue of the patients. The results were as follows : 1. The CYFRA 21-1 concentration in the cancer boundary, the most malignant region,(348.6+/-89.9 ng/ml) was the lowest value. The CYFRA 21-1 concentration in unaffected tissue,(718.4+/-77.8 ng/ml) was higher than that in the main lesion. which had intact cellularity. 2. The EGF-R concentration in the main lesion was higher than that in the unaffected tissue, and the EGF-R concentration in a squamous cell cacinoma was higher than that in an adenocarcinoma. also, the EGF-R concentration in the cancer boundary was highest at stage I, II. The EGF-R concentration was higher in the main cancer lesion that in the unaffected tissue at stage III, IV. 3. The CYFRA 21-1 was a cytoplasmic skeleton and the EGF-R was a cell-wall component; there was no correlation. In conclusion, CYFRA 21-1 was abundant in the cytoplasm but had a higher concentration in the unaffected tissue than in the main cancer lesion. The CYFRA 21-1 concentration of the tissue did not reflect the amount of cancer activity, the EGF-R was located in the cell membrane, the level of tissue that reflects cancer activity, so the main cancer lesion had a higher concentration than the unaffected tissue. CYFRA 21-1 is not a useful tumor maker at the tissue level. Because the EGF-R concentration reflected the cancer activity, its a useful tumor marker for lung cancer.

Telomerase Activity in Non-small Cell Lung Cancer / 대한흉부외과학회지

Although many reseraches have been persued to detect the molecular tumor marker to define the cancer, ideal tumor marker which speak for the characteristics of malignancy and has high sensitivity and specificity is not known. One of the characteristics of the malignant cells is indefinite proliferative potential, in other word, immortality. The expression of telomerase and stabilization of telomeres are concomitant with the attainment of immortality in tumor cells; thus the measurement of telomerase activity in clinically obtained tumor samples may provide important information which would be useful as a diagnostic marker to detect immortal cancer cells. Telomerase activity was analyzed in 12 non-small cell lung cancer cell lines and 41 primary non-small cell lung cancers with the use of a PCR-based assay. All the cell lines and the majority of tumors displayed telomerase activity, but telomerase was not detectable in most of the corresponding pathologically-normal tissues. Telomere length was not correlated with telomerase activity. The present study indicate that measurement of telomerase activity may be useful as a molecular tumor marker in non-small cell lung cancer.