UROKINASE-TYPE PLASMINOGEN ACTIVATOR,ITS RECEPTOR AND INHIBITOR EXPRESSION IN HEPATOCELLULAR CARCINOMA RELATION TO CANCER INVASIVENESS AND PROGNOSIS

Objective: To study the relevance of uPA, uPAR and PAI 1 to hepatocellular carcinoma (HCC) Methods: The expression at protein level of uPA, uPAR and PAI 1 was determined in 48 cases of HCC and 12 cases of benign tumors of liver (as control) by immunohistochemistry Results: When compared to cancer adjacent liver tissue and the control, positive rate of immune staining for uPA, uPAR and PAI 1 on cell membrane were significantly higher in HCC cells ( P <0 05) Positive staining of uPA and uPAR was seen in 16 of 22 and 19 of 22 cases of HCC with invasion, respectively ( P <0 01 and P <0 001) In 8 of 8 cases with cancer embolus, and in 6 of 6 cases with lymph node metastasis was the expression of positive uPAR Compared with 2 of 17 cases without recurrence, uPAR was positive in 15 of 17 recurrent cases ( P <0 01) In 36 cases who survived, 17 was positive uPAR and 15 positive PAI 1, while in 12 cases who died 2 years after surgery, 12 were positive for uPAR and 9 positive PAI 1, respectively ( P <0 01 and P <0 05) In 15 positive cases for all three parameters, 11 had cancer invasion and 7 died within 2 years, while in negative cases, 2 had invasion and none died within 2 years ( P <0 05) Conclusion: Expression of uPA, uPAR and PAI 1 is increased in HCC, uPA and uPAR may contribute significantly to HCC invasion and metastasis uPAR and PAI 1 are associated with poor prognosis of HCC

Nomograms predicting long-term survival in patients with invasive intraductal papillary mucinous neoplasms of the pancreas: A population-based study

BACKGROUND There are few effective tools to predict survival in patients with invasive intraductal papillary mucinous neoplasms of the pancreas.AIM To develop comprehensive nomograms to individually estimate the survival outcome of patients with invasive intraductal papillary mucinous neoplasms of the pancreas.METHODS Data of 1219 patients with invasive intraductal papillary mucinous neoplasms after resection were extracted from the Surveillance,Epidemiology,and End Results database,and randomly divided into the training(n=853)and the validation(n=366)cohorts.Based on the Cox regression model,nomograms were constructed to predict overall survival and cancer-specific survival for an individual patient.The performance of the nomograms was measured according to discrimination,calibration,and clinical utility.Moreover,we compared the predictive accuracy of the nomograms with that of the traditional staging system.RESULTS In the training cohort,age,marital status,histological type,T stage,N stage,M stage,and chemotherapy were selected to construct nomograms.Compared with the American Joint Committee on Cancer 7th staging system,the nomograms were generally more discriminative.The nomograms passed the calibration steps by showing high consistency between actual probability and nomogram prediction.Categorial net classification improvements and integrated discrimination improvements suggested that the predictive accuracy of the nomograms exceeded that of the American Joint Committee on Cancer staging system.With respect to decision curve analyses,the nomograms exhibited more preferable net benefit gains than the staging system across a wide range of threshold probabilities.CONCLUSION The nomograms show improved predictive accuracy,discrimination capability,and clinical utility,which can be used as reliable tools for risk classification and treatment recommendations.

Advances in BRAF gene mutations in papillary thyroid carcinoma

Thyroid cancer is the most common endocrine system tumor.Ultrasound guided fine needle puncture(FNA)can identify benign and malignant thyroid nodules.However,due to the limitation of cytological detection,some thyroid nodules are difficult to distinguish benign and malignant.BRAF gene mutation is a common human oncogenic mutation and the highest mutation frequency in papillary thyroid carcinoma.The combination of FNA and BRAF gene detection can significantly improve the diagnostic rate of benign and malignant thyroid nodules and make up for the deficiency of single diagnosis of cytology.Moreover,while the incidence of thyroid cancer is growing rapidly worldwide,its mortality remains stable.The problem of overdiagnosis and overtreatment of thyroid cancer is becoming more and more obvious.However,due to the limitations of current studies on BRAF genes,its prognostic value for papillary thyroid carcinoma remains controversial.Therefore,in order to reduce the adverse effects of overdiagnosis and treatment,the relationship between gene and tumor biological behavior needs further study in the future.

TIMP-3 gene transfection suppresses invasive and metastatic capacity of human hepatocarcinoma cell line HCC-7721

BACKGROUND: Tissue inhibitor of metalloproteinases (TIMPs) can restrain the tumor growth by their protein property and matrix metalloproteinases (MMPs) inhibition. There are currently four known TIMP family members-TIMP-1, 2, 3, 4. We determined whether increasing levels of TIMP-3 expression could suppress the malignant phenotype of human hepatocarcinoma cell line HCC-7721. METHODS: A recombinant expression vector, which contained full-length cDNA of human TIMP-3, was constructed and transfected into the human hepatocarcinoma cell line HCC-7721 by the lipofectamine technique. In vitro and in vivo tests such as Western blotting, immunohistochemistry as well as xenografting in nude mice were used to analyze expression levels of TIMP and MMP, and changes in malignant phenotype after the gene transfection. RESULTS: TIMP-3 expression in TIMP-3 gene-transfected HCC-7721 cells was upregulated as assessed by Western blotting. The ability of in vitro invasion through a Boyden chamber was significantly decreased compared to controls. Following subcutaneous injection into nude mice, the TIMP-3 transfected cells suppressed primary tumor growth, as characterized by reduced tumor weight, size and microvasculature as well as maintaining the extracellular matrix. CONCLUSION: The results suggest that upregulation of TIMP-3 expression in HCC-7721 cells inhibits invasion capacity in vitro as well as tumorigenic and metastatic potential in nude mice.

PROXIMAL BRONCHIAL INVASION OF LUNG CANCER: ACLINICOPATHOLOGICAL STUDY

Objective: To investigate the characteristics of proximal bronchial invasion of lung cancer for different types. Methods: Proximal bronchus of 151 operatively resected specimens of hilar type lung cancer were selected for cross-sectional pathological study. Forty-one specimens were obtained from total pulmonectomy, and 110 from pulmonary lobectomy. Results: Evidence showed that the direct invasion of tumor cells through the submucosal layer or multiple layers was the major form of lung cancer spread; 96.6% of the cancerous invasion occurred at the proximal bronchial wall less than 1.5 cm apart from the margin of the cancer. The extension of invasion was correlated with the histopathologic type of cancer, mode of invasion and TNM classification (pT, pN). Besides, the invasion in the bronchial wall by metastatic lymph nodes was also an important way for the cancer to spread. Conclusion: In order to achieve the radical removal of a tumor, it is imperative to keep a distance of 1.5 cm or more between the excision margin of the bronchus and the tumor, and to completely resect the hilar and mediastinal lymph nodes.

Cell softness reveals tumorigenic potential via ITGB8/AKT/glycolysis signaling in a mice model of orthotopic bladder cancer

Background:Bladder cancer,characterized by a high potential of tumor recurrence,has high lifelong monitoring and treatment costs.To date,tumor cells with intrinsic softness have been identified to function as cancer stem cells in several cancer types.Nonetheless,the existence of soft tumor cells in bladder tumors remains elusive.Thus,our study aimed to develop a microbarrier microfluidic chip to efficiently isolate deformable tumor cells from distinct types of bladder cancer cells.Methods:The stiffness of bladder cancer cells was determined by atomic force microscopy(AFM).The modified microfluidic chip was utilized to separate soft cells,and the 3D Matrigel culture system was to maintain the softness of tumor cells.Expression patterns of integrinβ8(ITGB8),protein kinase B(AKT),and mammalian target of rapamycin(mTOR)were determined by Western blotting.Double immunostaining was conducted to examine the interaction between F-actin and tripartite motif containing 59(TRIM59).The stem-cell-like characteristics of soft cells were explored by colony formation assay and in vivo studies upon xenografted tumor models.Results:Using our newly designed microfluidic approach,we identified a small fraction of soft tumor cells in bladder cancer cells.More importantly,the existence of soft tumor cells was confirmed in clinical human bladder cancer specimens,in which the number of soft tumor cells was associated with tumor relapse.Furthermore,we demonstrated that the biomechanical stimuli arising from 3D Matrigel activated the F-actin/ITGB8/TRIM59/AKT/mTOR/glycolysis pathways to enhance the softness and tumorigenic capacity of tumor cells.Simultaneously,we detected a remarkable up-regulation in ITGB8,TRIM59,and phospho-AKT in clinical bladder recurrent tumors compared with their non-recurrent counterparts.Conclusions:The ITGB8/TRIM59/AKT/mTOR/glycolysis axis plays a crucial role in modulating tumor softness and stemness.Meanwhile,the soft tumor cells become more sensitive to chemotherapy after stiffening,that offers new insights for hampering tumor progression and recurrence.

Activated vascular endothelia regulate invasion of glioma cells through expression of fibronectin

Background Previous researches have indicated that glioma invasion may occur within a tumor-host microecology, and that fibronectin may be involved in glioma invasion as an important component of the extracellular matrix. However, how the interaction between tumor cells and vascular endothelial cells affects glioma invasion is poorly understood. The aim of this study was to investigate the effects of the interaction between tumor cells and vascular endothelial cells on glioma invasion, and the relationship of this interaction to fibronectin. Methods The localization of fibronectin in different brain astrocytoma tissues was determined by immunohistochemistry Then, vascular endothelial cells and glioma cells were co-cultured in a Transwell co-culturing system. Fibronectin expression was detected by reverse transcriptase-polymerase chain reaction, immunocytochemistry, and enzyme-linked immunosorbent assay. Additionally, the influence of the interaction between tumor cells and vascular endothelial cells on glioma cell invasion was determined by an in vitro rapid invasion test. Results In brain astrocytoma tissues, fibronectin was present on the endothelial cells, in the extracellular matrix. Fibronectin expression was greater in higher grade tumors than in lower grade tumors. The interaction of glioma cells and vascular endothelial cells in vitro induced fibronectin release from vascular endothelial cells, which in turn stimulated glioma cell migration. This effect was inhibited by fibronectin blocking antibody. Conclusion Glioma cells may induce vascular epithelial cells to express fibronectin, and in turn fibronectin could promote glioma cell invasion.

Original article——Surgical treatment of IIIb-T4 lung cancer invading left atrium and great vessels

Background IIIb-T4 non-small cell lung cancer （NSCLC） is commonly considered a contraindication to surgery, although chemo-radiotherapy also achieves a poor survival rate. We reviewed our experience with T4 NSCLC patients who underwent surgery to explore the indications and prognostic factors of surgical treatment of lung cancer invading the left atrium and great vessels. Methods We investigated a cohort of 105 patients, 79 men and 26 women, who underwent surgery from May 1996 to July 2008. Their pathological staging was T4No-2M0. The median age was 59 years, ranging from 36 to 75 years. Patients were grouped based on invading sites： tumors invading the left atrium （LA group）, tumors invading the superior vena cava （SVC group）, and tumors invading the intrapericardial pulmonary artery （PA group）. Patients were further characterized based upon the type of operation, complete resection and incomplete resection groups, and on the lymph node pathological status, No, N1 and N2 groups. We calculated the overall five-year survival rate. Results All patients received resection of primary lesions, with partial resection of the left atrium in the LA group （n=-25）, angioplasty of superior vena cava in the SVC group （n=-23） and intrapericardial ligation of the pulmonary artery in the PA group （n=57）. Complete resection was possible in 77 patients （73.3%）. The overall survival rate of the 105 patients was 41.0% at 5 years; 36.0% for the LA group, 34.8% for the SVC group and 45.6% for the PA group. Pathological N status significantly influenced the overall 5-year survival rate; 61.5% for No, 51.1% for the N1 and 11.8% for the N2 groups （N2 group versus No group, P 〈0.0001, N2 versus N1 group, P〈0.0001）. Surgical resection also influenced survival; 49.4% for the complete resection group and 17.9% for the incomplete resection group （P 〈0.0001）. Cox regression analysis demonstrated that pathological N status was a significant independent predictor of prognosis. Conclusions Pathological N status is a significant independent predictor for survival of patients with IIIb-T4 lung cancer invading the left atrium and great vessels. The completeness of resection has a significant influence on the overall 5-year survival rate. Surgery for T4 lung cancer may be effective in patients without mediastinal lymph node involvement.

Macrophage migration inhibitory factor enhances neoplastic cell invasion by inducing the expression of matrix metalloproteinase 9 and interleukin-8 in nasopharyngeal carcinoma cell lines

Background Nasopharyngeal carcinoma (NPC) shows highly invasive and metastatic features. This study aims to investigate macrophage migration inhibitory factor (MIF)-induced invasion of NPC cells in vitro and the effects on matrix metalloproteinases (MMPs) and interleukin-8 (IL-8),and to study the mechanism of tumor cell invasion and metastasis in the early stage of NPC. Methods Two nasopharyngeal carcinoma cell lines,CNE-1 and CNE-2,were adopted in this study. The NPC cell invasion and migration were evaluated by microinvasion assay. The variation of expression percentages of MMP2- or MMP9-positive cells was detected by flow cytometry in two cell lines with or without MIF treatment. Western blotting and RT-PCR were used to assay the protein and mRNA expressions of MMP2 and MMP9. The IL-8 concentration secreted by NPC cells was compared with the cells with different treatments using ELISA. Results After treating with MIF for 48 hours,the cell numbers of CNE-1 and CNE-2 which went through the 8-μm filter membrane were increased. Compared with non-MIF treated NPC cells,significant difference could be found both in CNE-1( P =0.005) and CNE-2 cells ( P =0.001) . The percentages of MMP9-positive cells were significantly increased in both CNE-1 [from (28.5±2.5)% to (82.4±3.5)%, P =0.001] and CNE-2 [from (32.8±3.5)% to (86.1±1.6)%, P =0.002]. The relative intensity of MMP9 protein expression was also enhanced in both cell lines (CNE-1: from 83.1±6.0 to 242.9±22.9, P =0.002;CNE-2: from 84.4±4.3 to 278.9±29.7, P =0.003). Correspondingly,the increased MMP9 mRNA expression level was significantly detectable in both cell lines.The concentration of IL-8 in the supernatant of CNE-2 was higher [(1201.8±593.3) pg/ml] after treatment. It was also remarkably higher than that in the supernatant of CNE-2 without treatment ( P =0.026). However,there was no significant difference in the concentration variation of IL-8 in CNE-1 ( P =0.581), while the IL-8 mRNA level was only enhanced in CNE-2. Conclusions MIF can induce potent invasion of NPC cell lines in vitro , and the infiltrating lymphocytes in NPC might be responsible for the invasion and metastasis of tumor cells. MIF cytokine which is secreted by these infiltrating lymphocytes might contribute to the invasion as well as metastasis of NPC in the early stages by induction of MMP9 and IL-8 in an indirect pathway.

Relations of proliferative activities of gastric carcinoma cells to lymphatic involvement, venous invasion and prognosis

Background This study was to evaluate bivariate bromodeoxyuridine(BrdUrd)/DNA flow cytometric analysis in detection of gastric carcinoma and to study the relations of cellular BrdUrd labeling indices (LI), G_2/M-phase fraction(G_2/MPF) and DNA ploidy pattern to lymphatic involvement, venous invasion and prognosis.Methods Fresh tumor samples from 60 patients with gastric carcinoma were analyzed by bivariate BrdUrd/DNA flow cytometry. The results were correlated with lymphatic vessel invasion, lymphatic node metastasis, the number of matastatic lymphatic nodes, and venous invasion. Propidium iodide (PI) was used as a fluorescent probe for total cellular DNA, and a monoclonal antibody against BrdUrd was used as a probe for BrdUrd incorporated into DNA. Fluorescent-labeled goat anti-mouse antibody was used as a second antibody. S-phase fractions were measured by in vitro BrdUrd labeling, and DNA ploidy and G_2/MPF were also measured. Comparison of survival was performed with the log-rank test, the Chi-square test for qualitative data, and Student’s t test for quantu data. Results BrdUrd LI and G_2/MPF values were significantly higher in tumors with lymphatic vessel invasion than in those without invasion respectively (P<0.01); the patients who had tumors with lymphatic vessel invasion showed a significantly poor prognosis (P<0.01). Both BrdUrd LI and G_2/MPF values were significantly higher in tumors with lymphatic node metastasis than in those without metastasis (P<0.01). A statistical significant difference was noted in the 5-year survival rates between the patients with lymph node metastasis and those without metastasis. Compared with diploid carcinoma, the incidence of lymph node metastasis was significantly higher in aneuploid carcinoma (P<0.05), and the patients with aneuploid carcinoma showed a significantly poor prognosis (P<0.05). BrdUrd LI was significantly higher in patients with more than 5 metastatic lymph nodes than those with 1-4 metastatic lymph nodes (P<0.05) and those without metastasis (P<0.01). G_2/MPF values in those patients either with more than 5 metastatic lymph nodes or 1-4 metastatic lymph nodes were higher than those without metastasis (P<0.01 and P<0.05). A statistical significance was seen in the 5-year survival rates among the patients with no metastatic lymph node, 1-4 metastatic nodes and more than 5 metastatic nodes (P<0.01). G_2/MPF values were significantly higher in patients with venous invasion than in those without invasion (P<0.01).Conclusions Positive correlations exist between cellular BrdUrd LI, G_2/MPF with lymphatic involvement and prognosis, and DNA aneuploid with lymphatic involvement and prognosis. The same was true between G_2/MPF value and venous invasion in gastric carcinoma.