IGF-1R and Bmi-1 expressions in lung adenocarcinoma and their clinicopathologic and prognostic significance.

IGF-1R and Bmi-1 play a critical role in cancer growth and survival. We explored the correlation between IGF-1R and Bmi-1, as well as their relationship with clinicopathological parameters and their impacts on outcomes in patients with lung adenocarcinoma resected. Tumors from 178 surgical lung adenocarcinoma patients were evaluated for IGF-1R and Bmi-1 expression by means of immunohistochemistry. The clinicopathological implications of these molecules were analyzed statistically. There was a significant correlation between the expression of IGF-1R and Bmi-1 (p = 0.011). The 5-year survival rate of patients with Bmi-1 positive was only 31.2%, but patients with Bmi-1 negative had a survival rate of 50.7% (p = 0.004). The pattern of survival curves showed that Bmi-1 was a significant prognostic factor of poor overall survival in lung adenocarcinoma patients. However, there was no obvious correlation between IGF-1R expression and patient survival. The results of multivariate Cox analysis revealed that the pathological stages and Bmi-1 expression were independent prognostic factors. Therefore, Bmi-1 may be a good biomarker to predict the prognosis of patients with completely resected lung adenocarcinoma.

A classification method based on principal components of SELDI spectra to diagnose of lung adenocarcinoma.

PURPOSE: Lung cancer is the leading cause of cancer death worldwide, but techniques for effective early diagnosis are still lacking. Proteomics technology has been applied extensively to the study of the proteins involved in carcinogenesis. In this paper, a classification method was developed based on principal components of surface-enhanced laser desorption/ionization (SELDI) spectral data. This method was applied to SELDI spectral data from 71 lung adenocarcinoma patients and 24 healthy individuals. Unlike other peak-selection-based methods, this method takes each spectrum as a unity. The aim of this paper was to demonstrate that this unity-based classification method is more robust and powerful as a method of diagnosis than peak-selection-based methods. RESULTS: The results showed that this classification method, which is based on principal components, has outstanding performance with respect to distinguishing lung adenocarcinoma patients from normal individuals. Through leaving-one-out, 19-fold, 5-fold and 2-fold cross-validation studies, we found that this classification method based on principal components completely outperforms peak-selection-based methods, such as decision tree, classification and regression tree, support vector machine, and linear discriminant analysis. CONCLUSIONS AND CLINICAL RELEVANCE: The classification method based on principal components of SELDI spectral data is a robust and powerful means of diagnosing lung adenocarcinoma. We assert that the high efficiency of this classification method renders it feasible for large-scale clinical use.

Expression of GPC3 protein and its significance in lung squamous cell carcinoma.

The purpose of this study was to investigate GPC3 gene expression in lung squamous cell carcinoma tissue and its correlation with clinical and tumor characteristics. Using RT-PCR, the presence of GPC3 gene expression was detected in cancer tissue and adjacent normal tissue in 66 cases of lung squamous cell carcinoma and positive rates were calculated. Using Western blot, changes in GPC3 protein expression were detected in lung squamous cell carcinoma and adjacent normal tissues. The percentage of tissue samples expressing GPC3 mRNA was significantly higher in lung squamous cell carcinoma than in adjacent normal tissue (P < 0.05). This percentage was also significantly higher for cases with lymph node metastasis than for those without lymph node metastasis (P < 0.05). Further, the percentage of samples expressing GPC3 mRNA was higher with lowering degrees of tumor differentiation (P < 0.05). Rates of GPC3 expression were, however, independent of patient gender, age, and tumor size (P > 0.05). The expression of GPC3 protein in lung squamous cell carcinoma was significantly higher than that in adjacent normal tissues (P < 0.05). The expression in cases with lymph node metastasis was significantly higher than in those without lymph node metastasis (P < 0.05), and GPC3 protein expression increased with lowering degrees of tumor differentiation (P < 0.05). Further investigation is warranted for the association of initiation, development, invasion, and metastasis of disease.

[Correlations between IGF-IR expression and clinicopathological factors and prognosis in patients with lung adenocarcinoma].

BACKGROUND AND OBJECTIVE: the incidence of lung adenocarcinoma increases rapidly, and IGF-IR is the key mediator of several growth factors signal transduction, therefore it plays an important role in the proliferation and differentiation of cancer cell. The aim of this study is to detect the expression of IGF-IR in lung adenocarcinoma and to evaluate its implication for the clinicopathological factors and prognosis of patients with this disease. METHODS: the IGF-IR expression was detected by immunohistochemical staining. Correlations between IGF-IR expression with clinicopathological factors were analyzed using the Chi-squared test. The Kaplan-Meier method was used to calculate the overall patient survival rate, and the difference in survival curves was evaluated using a Log-rank test. Univariate and multivariate analysis was carried out using the Cox proportional-hazard model. RESULTS: in 126 cases of tumor sections tested, IGF-IR were detected in 89 cases. Statistical analysis revealed that the IGF-IR expression was related to tumor size and T stage, while there were no relations between IGFIR expression and age, gender, smoking, pathological stages, and differentiation. Cox analysis indicated that metastasis and chemotherapy efficacy were the prognostic factors in these patients, while IGF-IR expression was not the independent prognostic factor. CONCLUSIONS: the IGF-IR expression is related to tumor size and T stage, while there is no relation between IGF-IR expression and prognosis.

[The application of endobronchial ultrasound-guided transbronchial needle aspiration: report of 70 cases].

OBJECTIVE: To evaluate the diagnostic yield and the safety of endobronchial ultrasound-guided transbronchial needle biopsy (EBUS-TBNA) in mediastinal and hilar lymph nodes and lung tumors. METHODS: EBUS-TBNA was performed in 70 patients with thoracic masses or mediastinal-hilar lymphoadenopathy proved by CT scan. RESULTS: From July 2009 to January 2010, 70 patients were included in the study. EBUS-guided TBNA was performed to obtain samples from mediastinal and hilar lymph nodes (120 stations) and lung tumors (11 masses). In 46 cases of newly diagnosed lung cancer, 44 were confirmed by EBUS-TBNA without on site cytology assistance, with 2 false negative cases. The sensitivity, specificity, positive predictive value, negative predictive value, and accuracy of EBUS-TBNA in the diagnosis of lung cancer were 96%, 100%, 100%, 92% and 97% respectively. Non-caseous granuloma formed by epithelioid cells was found in EBUS-TBNA histological specimen from 5 out of 10 patients with clinically diagnosed sarcoidosis. TBNA cytological smear showed acid-fast bacilli and histology of the lymph node demonstrated coagulatory necrosis from 1 out of 4 tuberculous cases. The procedure was uneventful, and there were no complications. CONCLUSION: EBUS-TBNA is an effective and safe method for the diagnosis of bronchogenic carcinoma and unknown mediastinal-hilar lymphadenopathy.

The expression of stem cell-related indicators as a prognostic factor in human lung adenocarcinoma.

INTRODUCTION: The purpose of the present study was to detect the presence of BASC-like stem cell-related indicators, such as clara cell secretory protein (CCSP), Octamer-4 (OCT4) and Bmi-1, and evaluate their implications in the prognosis of patients with lung adenocarcinoma. METHODS: Specimens of 134 cases of lung adenocarcinoma were collected after radical surgery from January 1999 to June 2004. RESULTS: One hundred and twenty-six cases showed cells that were positive for CCSP, 99 cases positive for OCT4, 91 cases simultaneous expression of CCSP and OCT4 and 74 cases positive for Bmi-1. Bmi-1 was significantly higher in patients at stage III compared to patients at stages I and II. The pattern of survival curves showed that Bmi-1 was a significant prognostic factor of poor overall survival in lung adenocarcinoma patients (P = 0.0000), and the patients with OCT4(+) expression showed a greater increase in mortality than OCT4(-) patients (P = 0.0103). The results of univariate and multivariate Cox analysis revealed that the pathological stages of tumor node metastases (P = 0.037), OCT4 (P = 0.046) and Bmi-1 expression (P = 0.001) were independent prognostic factors. CONCLUSIONS: OCT4 and Bmi-1 may be good biomarkers to predict the prognosis of patients with completely resected lung adenocarcinoma.

[Effects of a protein kinase C inhibitor combined with cisplatin on non-small cell lung cancer].

OBJECTIVE: To study the effects and possible mechanisms of the protein kinase C (PKC) inhibitor chelerythrine chloride (CH), combined with cisplatin (DDP) on human non-small cell lung cancer. METHODS: The effect of CH, DDP and the combination on proliferation and apoptosis of human lung cancer cell line A549 were evaluated by MTT assay and flow cytometry respectively. The inhibitory effects of CH and DDP on neoplasia were verified on subcutaneous implanted tumor of nude mice. Implanted tumor models were constructed in nude mice using human lung adenocarcinoma cell line A549. Twenty-four BALB/c nude mice with implanted tumors were divided into 4 groups randomly: group CH, group DDP, group CH + DDP, and normal saline group (group NS), each with 5 mice. CH, DDP or NS were intraperitoneally injected into nude mice for 3 weeks (DDP or NS was injected once a week, CH was injected twice a week). RESULTS: CH inhibited A549 cell proliferation in a concentration-dependent pattern. When CH and DDP were combined, the inhibitory effect was enhanced in a synergistic or additive pattern. Both CH and DDP significantly increased the apoptosis of A549 cells, and this action was remarkably increased when DDP was combined with CH. CH and DDP inhibited the growth of subcutaneous implanted tumor in nude mice and the inhibitory rate of group CH + DDP (80.5%) was significantly higher than that of group CH (72.4%) or group DDP (64.3%) (t = 11.34, P < 0.01). CONCLUSION: CH combined with DDP shows significantly synergistic anti-tumor effects on non-small cell lung cancer cell line A549 and subcutaneous implanted tumor in nude mice, possibly by enhancement of growth inhibition and apoptosis induction on tumor cells.

Prognostic impact of vascular endothelial growth factor-A and E-cadherin expression in completely resected pathologic stage I non-small cell lung cancer.

OBJECTIVE: The purpose of this study was to evaluate the value of vascular endothelial growth factor-A and E-cadherin expression as well as other confirmed prognostic factors in predicting the clinical outcome after definitive surgery of pathologic stage I non-small cell lung cancer. METHODS: One hundred and eighty-five consecutive and non-selected patients who underwent definitive surgery for stage I non-small cell lung cancer in our institute were included in this study. Formalin-fixed paraffin-embedded specimens were stained for vascular endothelial growth factor-A and E-cadherin and the correlation between the staining, its clinicopathological parameters and its prognostic power were analyzed statistically. RESULTS: Of the 185 patients studied, 92 cases (49.7%) were strongly positive for vascular endothelial growth factor-A. Vascular endothelial growth factor-A expression was only related to visceral pleural involvement (P < 0.001). A total of 95 carcinomas (51.4%) were E-cadherin-negative tumors. E-cadherin expression correlated with histology (P < 0.001), tumor size (P = 0.001) and visceral pleural involvement (P < 0.001). In univariate analysis by log-rank test, gender, tumor size, lymphovascular invasion, visceral pleural involvement, vascular endothelial growth factor-A expression and E-cadherin expression were significant prognostic factors (P = 0.003, 0.042, 0.026, 0.035, 0.008 and 0.006, respectively). In multivariate analysis, gender, vascular endothelial growth factor-A and E-cadherin expression maintained its independent prognostic influence on overall survival (P = 0.013, <0.001 and 0.036, respectively). CONCLUSIONS: Expression of vascular endothelial growth factor-A is related to visceral pleural involvement, and E-cadherin expression correlates with histology, tumor size and visceral pleural involvement. Multivariate analysis confirmed gender, vascular endothelial growth factor-A and E-cadherin expression were significant predictive factors for overall survival in completely resected pathologic stage I non-small cell lung cancer.

Inducible costimulator (ICOS) enhances the cytolytic activity of cytokine-induced killer cells against gallbladder cancer in vitro and in vivo.

The generation of genetically modified immunological effector cells is of potential therapeutic value in the treatment of malignancies. Cytokine-induced killer cells (CIKs) have been described as highly efficient cytotoxic effector cells capable of recognizing and lysing tumor cell targets in a non-major histocompatibility complex restricted fashion. In the present study, we evaluated the effects of inducible costimulator (ICOS) on the cytotoxicity of CIK cells against gallbladder cancer. We first prepared CIK-ICOS cells by the transfection of ICOS genes into induced CIK cells, whereas untransfected or enhanced green fluorescent protein (EGFP)-transfected CIK cells were treated as controls. We found that CIK-ICOS cells displayed better proliferation and lower apoptosis than the other two CIK control cells after culture. The interferon-gamma level in the culture supernatant of CIK-ICOS cells was also significantly elevated, compared to CIK or CIK-EGFP cells. The cytotoxic effect of CIK-ICOS cells against gallbladder cancer cells was dramatically enhanced at the E:T ratio of 20:1, compared to that of CIK or CIK-EGFP cells. When injected into gallbladder tumor-bearing SCID mice, CIK-ICOS cells significantly slowed down the growth rate of xenografts. CIK-ICOS-treated mice exhibited the least volume variation of the xenografts and more severe necrosis, compared to saline, CIK, or CIK-EGFP cell-treated mice, accompanied by a better in situ survival around xenografts than the other two control cells. Taken together, our results demonstrated that ICOS could enhance the cytotoxic activity of CIK cells, in part, by augmenting cytokine secretion and prolonging cell survival both in vitro and in vivo. This might be considered as the potential immunomodulator for clinical therapy.

[Prognostic Analysis of ERCC1, RRM1 and p53 Expressions in Postoperative Stage I-II Lung Cancer.].

BACKGROUND: Cisplatin is an important drug in lung cancer chemotherapy. It has been proven that ERCC1, RRM1, p53 expressions were related to resistance to platinum and prognosis of the patcents with lung cancer. The aim of this study is to analyze the association of the expression of ERCC1, RRM1, p53 with postoperative survival in patients with stage I-II non-small-cell lung cancer (NSCLC), and to explore the relationship between the expression of ERCC1, RRM1, p53 and resistance to cisplatin. METHODS: A total of 75 patients with stage I-II NSCLC receiving radical resection from Feb. 1992 to Jan. 1994 were followed up. Postoperative patients with stage I were randomized two groups (chemo and non-chemo groups). All patients with stage II received adjuvant cisplatinbased chemotherapy. Immunohistochemical staining was used to detect the expression of ERCC1, RRM1, p53 in paraffinembedded specimens. RESULTS: In stage I NSCLC, the prognosis of the patients with high expression of ERCC1 (High-ERCC1) was better than those with low expression of ERCC1 (Low-ERCC1). 1, 3, 5-year survival rate in the patients with high expression of ERCC1 was 100.00%, 91.30%, 86.74% and in those with Low-ERCC1 was 96.43%, 60.71%, 57.14%, respectively (P =0.0058). The patients with High- ERCC1 had a better survival rate than those with Low-ERCC1 in stage I NSCLC without chemotherapy. MST in high and low expression of ERCC1 was 72.00(+) months and 64.67 months, respectively (P =0.0327). In contrary to stage I NSCLC, the patients with had a better survival rate than those with in stage II. MST was 60.00(+) months in stage II patients with low expression of ERCC1, but MST was only 25.50 months with (P =0.0442). The postoperative survival of NSCLC patients was not any statistical different between with high expression and low expression of RRM1 and p53. CONCLUSIONS: High expression of ERCC1 is a better independent prognostic factor in stage I NSCLC patients. Cisplatin-base chemotherapy prolongs survival in stage II NSCLC patients with. Adjuvant chemotherapy regimen is determined according to ERCC1 expression levels in resected NSCLC.

[Antitumor effects of specific cyclooxygenase inhibitors combined with chemotherapeutic agents on gastric cancer cells in vitro].

OBJECTIVE: To study the effects of two specific cyclooxygenase inhibitors (SCI), rofecoxib and celecoxib, combined with chemotherapeutic drugs 5-Fu, DDP and VP-16 on gastric cancer cell line BGC-823, and to evaluate whether specific cyclooxygenase inhibitors can be used as a synergetic agent in chemotherapy. METHODS: The gastric cancer cell line BGC-823 cells were incubated for 48 hours with rofecoxib and celecoxib, 5-Fu, DDP and VP-16 (concentration gradient of 5-Fu, DDP and VP-16:1 microg/ml, 10 microg/ml and 100 microg/ml), or in combination, respectively. MTT working solution was added to each culture and calculated the survival rates of gastric cancer cells. Median-effect principle and Professor Jin's evaluation methods were applied to detect the interaction between the specific cyclooxygenase inhibitors and chemotherapeutic agents. RESULTS: The inhibition rates of gastric cancer cells were 42.63% +/- 1.26% and 50.67% +/- 2.35% by treatment with 0.1 micromol/L rofecoxib and 50 micromol/L celecoxib, respectively. The inhibition rates of gastric cancer cells by treatment with 5-Fu, DDP and VP-16 at different concentrations (1 microg/ml, 10 microg/ml and 100 microg/ml) were 39.75% +/- 3.14%, 49.96% +/- 2.08%, 87.93% +/- 3.66%; 48.28% +/- 2.08%, 59.46% +/- 1.69%, 88.23% +/- 4.81%; and 29.23% +/- 3.27%, 49.34% +/- 3.75%, 79.24% +/- 2.44%, respectively. However, the inhibition rates showed a synergetic role while combined the two SCI (0.1 micromol/L rofecoxib and 50 micromol/L celecoxib) with chemotherapeutic agent at different concentrations (P <0.05). CONCLUSION: Both rofecoxib and celecoxib have an ability to suppress gastric cancer cells in vitro, and the synergetic role becomes evident when rofecoxib and celecoxib are combined with chemotherapeutic agents at different concentrations, which indicate that the two specific cyclooxygenase inhibitors may be used as a chemotherapeutic sensitizer.

Proteomic analysis on metastasis-associated proteins of human hepatocellular carcinoma tissues.

PURPOSE: A comparative proteomic approach was used to identify and analyze proteins related to metastasis of hepatocellular carcinoma (HCC). METHODS: Proteins extracted from 12 HCC tissue specimens (six with metastases and six without) were separated by two-dimensional gel electrophoresis (2-DE). The protein spots exhibiting statistical alternations between the two groups through computerized image analysis were then identified by mass spectrometry. In addition immunohistochemistry (IHC), Western blotting and RT-PCR were performed to verify the expression of certain candidate proteins. RESULTS: 16 proteins including HSP27, S100A11, CK18 were annotated by mass spectrometry, relevant to chaperone function, cell mobility, cytoskeletal architecture, respectively. Most were previously unconnected with metastasis of HCC. Of these HSP27 was found overexpressed consistently in 2-DE patterns of all metastatic HCC tissues compared with nonmetastatic ones. IHC and Western blotting of HCC tissues confirmed this difference while RT-PCR did not. CONCLUSION: There are various proteins joined together in HCC metastasis. The overexpression of HSP27 may serve as a biomarker for early detection and therapeutic targets unique to the metastatic phenotype of HCC.

[Proteomic analysis on metastasis-associated proteins of hepatocellular carcinoma tissues].

OBJECTIVE: A comparative proteomic approach was used to identify and analyze proteins relevant to metastasis of hepatocellular carcinoma (HCC). METHODS: Proteins extracted from 12 liver tumor tissue specimens (6 with metastases and 6 without) were separated by two-dimensional gel electrophoresis (2-DE). Comparative analyses of 2-DE protein patterns between the two groups were done using computerized image analysis. Selected proteins exhibiting statistically significant alternations were identified by mass spectrometry. Immunohistochemistry, Western blotting and RT-PCR were performed to examine the expressions of the candidate proteins. RESULTS: 16 proteins including HSP27, S100A11, CK18 were identified using mass spectrometry, which were related to cell mobility, signal transduction, and energy metabolism respectively. Of these, HSP27 was found to be uniquely over-expressed in 2-DE maps of all metastatic HCCs when compared to the non-metastatic HCC tissues. Immunohistochemistry and Western blotting of HCC tissues confirmed this difference while RT-PCR did not. CONCLUSION: There are different proteins working together that affect the metastasis of HCCs. The overexpression of HSP27 may serve as a biomarker for early detection and therapeutic targets to the metastatic phenotype of HCC. The role of HSP27 in HCC metastasis warrants further investigation.

[Efficient generation and functional characteristics of dendritic cells from malignant pleural effusion in patients with lung cancer].

OBJECTIVE: To clarify whether functionally competent dendritic cells (DC) can be generated from malignant pleural effusion in patients with lung cancer. METHODS: Malignant effusion-associated monocytes were separated by adherence from malignant effusion-associated mononuclear cells and cultured in medium with granulocyte macrophage colony-stimulating factor (GM-CSF) plus interleukin 4 (IL-4). TNF-alpha was added for the last 24 h before culture termination. Cultured DC were identified by (1) using microscopy, scanning electron microscopy, and immunocytochemistry for the morphological features; (2) phenotypic markers; and (3) functional characteristics including a high stimulatory capacity to activate proliferation of lymphocyte in an allogeneic mixed leukocyte reaction and the ability to produce high levels of IFN-gamma. RESULTS: Cultured DC had the typical morphological features. The phenotype of DCs generated from effusion showed higher expression of CD(86) (84.6 +/- 6.1)%, HLA-DR (81.1 +/- 13.0)%, CD(40) (42.0 +/- 21.7)%, CD(1a) (20.0 +/- 9.5)% and lower expression of CD(14) (4.8 +/- 3.5)% than the control group. There was a significant difference in the stimulatory activity in allogeneic lymphocyte proliferation and the ability to produce high levels of IFN-gamma between DC derived from the malignant effusion and the control group. CONCLUSION: These findings suggest that DC can be generated from malignant pleural effusion, which might be a useful source of DC for immunotherapy.

Human acyl-coenzyme A:cholesterol acyltransferase 1 (acat1) sequences located in two different chromosomes (7 and 1) are required to produce a novel ACAT1 isoenzyme with additional sequence at the N terminus.

A rare form of human ACAT1 mRNA, containing the optional long 5'-untranslated region, is produced as a 4.3-kelonucleotide chimeric mRNA through a novel interchromosomal trans-splicing of two discontinuous RNAs transcribed from chromosomes 1 and 7. To investigate its function, we express the chimeric ACAT1 mRNA in Chinese hamster ovary cells and show that it can produce a larger ACAT1 protein, with an apparent molecular mass of 56 kDa on SDS-PAGE, in addition to the normal, 50-kDa ACAT1 protein, which is produced from the ACAT1 mRNAs without the optional long 5'-untranslated repeat. To produce the 56-kDa ACAT1, acat1 sequences located at both chromosomes 7 and 1 are required. The 56-kDa ACAT1 can be recognized by specific antibodies prepared against the predicted additional amino acid sequence located upstream of the N-terminal of the ACAT1(ORF). The translation initiation codon for the 56-kDa protein is GGC, which encodes for glycine, as deduced by mutation analysis and mass spectrometry. Similar to the 50-kDa protein, when expressed alone, the 56-kDa ACAT1 is located in the endoplasmic reticulum and is enzymatically active. The 56-kDa ACAT1 is present in native human cells, including human monocyte-derived macrophages. Our current results show that the function of the chimeric ACAT1 mRNA is to increase the ACAT enzyme diversity by producing a novel isoenzyme. To our knowledge, our result provides the first mammalian example that a trans-spliced mRNA produces a functional protein.

Hematogenous dissemination of lung cancer cells during surgery: quantitative detection by flow cytometry and prognostic significance.

Shedding of neoplastic cells into the circulation is an essential event for the hematogenous metastasis of solid tumors. Recently, several studies reported that a high frequency of cancer cells could be detected in the bloodstream during surgery. The intraoperative detection of hematogenous dissemination of cancer cells was able to identify a subset of patients with malignant diseases at high risk for postoperative metastasis and to predict a poor prognosis. In order to evaluate the association between intraoperative dissemination of cancer cells and postsurgical survival of patients with non-small cell lung cancer (NSCLC), we developed a flow cytometric assay for specific detection of lung cancer cells in the blood. The monocyte-enriched population in the blood was separated by a modified Ficoll-Hypaque density centrifugation and then labeled with a combination of monoclonal antibodies specific for CD45, cytokeratin (CK) and two antigens expressed on lung cancer cells (2F7 and S5A). The assay could detect quantitatively lung cancer cells (defined as CD45(-1) CK(+) 2F7/S5A(+) cells), with the sensitivity limit of one cancer cell in 10(5) normal leukocytes. The specificity for lung cancer was 97%, which was calculated from the results of healthy subjects (20 cases) and patients affected with benign pulmonary diseases (26 cases) or esophageal cancer (14 cases). Blood samples of 31 NSCLC patients were collected from pulmonary vein during open thoracic surgery. Fifteen of them (48.4%) were found to have positive test results. The average cancer cell counts in these cases were 0.306 x 10(6)/l. Patients under 55 years of age had a significantly higher percentage of positive findings than those over 55 years of age (P < 0.05). The positive rate increased over the stages and lymph node status, but the differences were not statistically significant. Moreover, patients with squamous cell carcinoma at later stages (stages III and IV) had an increased frequency of positive test results than those at earlier stages (stages I and II, P < 0.05). In contrast, no such a difference was found in cases with adenocarcinoma. On the basis of 30-months follow-up date, the median survival time and 2-year survival rate for patients with positive and negative findings were 11 vs. 27 months, and 26.7 vs. 62.5%, respectively. There was a statistically significant difference between overall survival curves that favored the patients with negative test results (P = 0.023). Multivariate analysis indicated the stage of disease and the positive test results as two independent factors that affected survival time (P = 0.017 and 0.027). When a comparison was made within the patients at stages III and IV, the presence of cancer cells in blood was associated with a significantly shorter survival. These data indicate that the hematogenous dissemination of lung cancer cells during surgery would be one of the mechanisms of postoperative tumor metastasis. The detection of these cells may help to identify patients with poor prognosis.

[Vascular endothelial growth factor promotes hematogenous metastasis of cancer cells in patients with non-small cell lung cancer].

OBJECTIVE: To evaluate the effect of vascular endothelia1 growth factor (VEGF) on the hematogenous metastasis of non-small cell lung cancer (NSCLC). METHODS: The identification of lung cancer cells in the peripheral blood were carried out by cytological, immunohistocytologica1 and immunofluorecent stains respectively, following isolation of cytokeratin-expressing cells with magnetic activated cell sorting. The quantification of cancer cells in the blood was performed according to the established flow cytometric assay. The plasma VEGF was measured by commercially available ELISA kit. RESULTS: The lung cancer cells in the blood, showing a remarkable nuclear polymorphism, expressed the epithelial marker cytokeratin and telomerase reverse transcriptase (hTERT). These cells were stained positive by an NSCLC-specific monoclonal antibody S5Al0-2, but negative by antibodies against CD34 and CD45 antigens. Using the flow cytometric assay, 44 cases (28.6%) of l54 NSCLC patients were found to have cancer cells in their blood, with the incidence of positive cases correlated with the stage of disease. The plasma VEGF level was significantly increased in NSCLC patients in comparison with healthy individuals and patients with benign pulmonary diseases. This level was correlated with the stages of disease in patients with adenocarcinoma. In patiens with cancer cells in their blood, a higher level of plasma VEGF was related with an increased number of cancer cells. CONCLUSION: The plasma VEGF level is increased in NSCLC patients with approximate1y one fourth to have cancer cells in the peripheral blood. In these patients, increased VEGF level promotes hematogenous tumor metastasis, as indicated by a much higher number of cancer cells in the blood.

[The relationship between drug sensitivity and expression of drug resistance gene mutations in non-small cell lung cancer].

OBJECTIVE: To study the relationship between the expression of drug resistance genes and the results of drug sensitivity test. METHODS: Surgical or biopsy specimens from 48 patients with non-small cell lung cancer (NSCLC) were measured for drug sensitivity by Annexin V combined with PI using flow cytometry. The drug resistance genes MDR(1), GST-pi and MPR were measured by RT-PCR. The relationship between expression of drug resistance gene mutations and the drug sensitivity of lung cancer was analyzed. RESULTS: The anti-tumor cytotoxicity of MMC, DDP, VDS, NVB, TAX, GEM, VP-16 and VCR were measured, and their respective tumor inhibition rates were (10.3 +/- 17.1)%, (20.7 +/- 22.2)%, (5.6 +/- 14.9)%, (7.9 +/- 16.2)%, (15.7 +/- 21.8)%, (11.2 +/- 13.8)%, (9.7 +/- 20.1)%, and (4.7 +/- 8.7)%. The positive rates of MDR(1), MRP and GST-pi expression were 67% (32/48), 42% (20/48), and 48% (23/48) respectively. There was no association between the expression of drug resistance genes MRP and GST-pi and the pathology or the stage of lung cancer. Interestingly, the over-expression of MRP was related to drug resistance to NVB, VDS and MMC; while the over-expression of GST-pi was related to resistance to DDP. No relationship was found between MDR(1) over-expression and drug resistance. CONCLUSION: The expression of some drug resistance genes is related to drug sensitivity test. The detection of the genes may be clinically useful in the administration of chemotherapy.