Effect of Antisense Oligodeoxynucleotide of Vascular Endothelial Growth Factor C on Lymphangiogenesis and Angiogenesis of Pancreatic Cancer

In order to investigate the effect of antisense oligonucleotide (ASODN) of vascular en-dothelial growth factor C (VEGF-C) on lymphangiogenesis and angiogenesis of pancreatic cancer, antisense and scamble-sense oligonucleotide of VEGF-C were constructed, and the model of nude mice with orthotopically xenografted human pancreatic cancer cells (Panc-1) was established. Thirty nude mice were randomly divided into 3 groups: PBS control group (group A), scramble-sense con-trol group (group B) and antisense group (group C). All nude mice were treated once every 2 days as 3 times per week, for 3 weeks (oligonucleotide 10 mg/kg every time). After treatments were com-pleted, ELISA method was used to examine the concentration of VEGF-C in plasma and immunohis-tochemical method to examine microvessel density (MVD), lymphtic vessel density (LVD) of pan-creatic cancer. The results showed that the expression of VEGF-C was inhibited significantly in group C. The concentrations were 237.5±41.5, 221.5±52.3 and 108.6±14.9 pg/mL in groups A, B and C re-spectively (P<0.01). LVD in groups A, B and C was 13.8±2.1, 12.4±1.9 and 4.2±1.6 respectively (P<0.01). MVD in groups A, B and C was 27.5±8.7, 25.9±4.2 and 19.4±5.6 respectively with no sig-nificant difference among the groups (P>0.05). It was suggested that VEGF-C ASODN decreased the expression levels of VEGF-C in nude mice with orthotopically xenografted human pancreatic cancer, and it could inhibit lymphangiogenesis, but had no significant effect on angiogenesis.     

Expression and Clinical Implication of HIF-la and VEGF-C in Non-small Cell Lung Cancer

To study the expression and implication of HIF-la and VEGF-C in non-small cell lung cancer (NSCLC) and its relationship with clinical pathological features of NSCLC, immunohistochemical SP was used to detect the expression of HIF-1а and VEGF-C proteins in 48 NSCLC tissues and the same para-cancerous tissues. The positive rates of HIF-1а and VEGF-C were 70.8% (34/48) and 68.8% (33/48) respectively. The expression of HIF-la protein was detected in a significantly greater proportion in NSCLC carcinoma tissues than that in para-cancerous tissues (12.5% and 16.7%,P<0.05). The positive rates of HIF-1а and VEGF-C were correlated with lymph node metastasis and TNM stage. No relationship was found between the two factors and age, sex, pathological subtypes and histological grades. The positive rates between HIF-1а and VEGF-C were correlated (P<0.05).HIF-1а and VEGF-C were over-expressed in NSCLC. They may be involved in the carcinogenesis of NSCLC, and play an important role in invasion and metastasis of NSCLC. HIF-1а and VEGF-C work synergically in the process of NSCLC.     

Expression and Clinical Implication of HIF-1α and VEGF-C in Non-small Cell Lung Cancer

To study the expression and implication of HIF-1α and VEGF-C in non-small cell lung cancer （NSCLC） and its relationship with clinical pathological features of NSCLC, immunohisto-chemical SP was used to detect the expression of HIF-1α and VEGF-C proteins in 48 NSCLC tissues and the same para-cancerous tissues. The positive rates of HIF-1α and VEGF-C were 70.8% （34/48） and 68.8% （33/48） respectively. The expression of HIF-1α protein was detected in a significantly greater proportion in NSCLC carcinoma tissues than that in para-cancerous tissues （12.5% and 16.7%, P〈0.05）. The positive rates of HIF-1α and VEGF-C were correlated with lymph node metastasis and TNM stage. No relationship was found between the two factors and age, sex, pathological subtypes and histological grades. The positive rates between HIF-1α and VEGF-C were correlated （P〈0.05）. HIF-1α and VEGF-C were over-expressed in NSCLC. They may be involved in the carcinogenesis of NSCLC, and play an important role in invasion and metastasis of NSCLC. HIF-1α and VEGF-C work synergically in the process of NSCLC.     

Clinical Significance of VEGF-C and VEGFR-3 Expression in Non-small Cell Lung Cancer

The relationship between vascular endothelial growth factor-C (VEGF-C)/vascular en-dothelial growth factor receptor 3 (VEGFR-3) expression and clinicopathologic features of the patients with non-small cell lung cancer (NSCLC) was investigated. The expression of VEGF-C and VEGFR-3 was assessed in 65 patients with NSCLC by immunohistochemistry. The significance of VEGF-C and VEGFR-3 expression was analyzed statistically. The results showed that VEGF-C and VEGFR-3 were highly expressed in cytoplasm and membrane in lung cancer tissues with the positive rate being 55.4 % and 52.3 % respectively, while there was no expression in the normal lung tissues. The expression of VEGF-C was significantly increased in adenocacinoma as compared to other types of NSCLC (P<0.05). The VEGFR-3 expression was closely related with lymph node metastasis (P<0.01) and TNM stage (P<0.05). There was a positive correlation between the VEGF-C and VEGFR-3 expression in NSCLC patients (r=0.658, P<0.01). It is suggested that VEGFR-3 plays an important role in the lymphatic metastasis of NSCLC. The interaction between VEGF-C and VEGFR-3 may be deeply involved in the mechanism of lung cancer metastasis.     

The Expression of Hypoxia Inducible Factor 1-alpha in Lung Cancer and Its Correlation with P53 and VEGF

To investigate the expression of hypoxia inducible factor 1-alpha (HIF-1α) and its corre-lation with P53 and vascular endothelial growth factor (VEGF), immunohistochemical technique was employed to detect the protein expressions of HIF-1α, P53 and VEGF in specimens from 57 pa-tients with lung cancer. The results indicated that the total positive proportion of HIF-1α expres-sion was 63% and the HIF-1α expression was more frequent in bronchiole-alveolar carcinoma (86%) than in other lung cancer. There was a strong association of HIF-1α with VEGF and P53 pro-tein expressions. It is concluded that HIF-1α overexpression is a common event in lung cancer,which may be related to the up-regulation of the angiogenic factor VEGF and oncogene mutant P53 protein.     

Expression and significance of clusterin in normal prostate, benign prostate hyperplasia and prostate cancer

Objective To investigate the expression and significance of Qusterin in nonnal prostate, benign prostate hyperplasia(BPH) and prostate cancer. Methods Clusterin expression in samples of 12 normal prostate, 15 BPH.and 56 prostate cancer were studied by immunohistochanical stain. Results Of 83 cases, 67 are positive or weak positive (81 % ). The rate of positive or weak positive for normal prostate,BPH and prostate cancer was 17% (2/12),73% (11/15), and 96% (54/56) respectively. The expression level of Qusterin in prostate cancer was much higer than in normal prostate(t=8.82,P<0.01).BPH(t = 7.63,P< 0.01) was related positively with pathological grade(r = 0.649,P<0.01) and stage(r = 0.609, P< 0.01) of prostate cancer. Conclusion Qusterin may play an important role in the biological characteristics of prostate cancer by the antiapoptosis pathway. 9 refs,4 figs,1 tab.     

Hypoxia-Inducible Factor-1alpha Suppressing Apoptosis and Increasing Tolerance of Lung Cancer Cells to Chemotherapy

In order to construct plasmid of hypoxia-inducible factor-lalpha (HIF-1α), and transfect into human lung cancer cells A549, the change in sensitivity of lung cancer cells A549 to chemotherapy was observed. HIF-1αmRNA structure region was amplified by RT-PCR and inserted into plasmid pcDNA3. The expression plasmid pcDNA3/HIF-1αwas transfected into A549 with Lipofec-tAMINE?000. The expression of HIF-1αprotein was detected by Western blot. After A549 cells were transfected with HIF-1αprior to addition of 5-Fu, the growth activity was measured by growth curve, apoptosis was detected by flow cytometry at 48 h, and the levels of caspase3 and MDR-1 were determined by Western blot. The results showed that the constructed expression plasmid was analyzed with restriction enzymes and gel electrophoresis. Two DNA lanes at 2.55 kb and 5.4 kb respectively were found, which were consistent with that expected. The growth rate in 5-Fu group was significantly inhibited, and the apoptosis index and caspase3 activity were increased significantly as compared with control group. After HIF-1αbeing transfected into A549, the activity of MDR-1 was increased and the effect of 5-Fu was weakened. In conclusion, HIF-1αcan promote chemoresistance by increasing the activation of MDR1 and suppressing apoptosis during lung cancer cells A549 induced with 5-Fu.     

Prognostic significance of lymph node metastasis in surgical resection of esophageal cancer.

OBJECTIVE Although surgery is relatively successful in eradicating local tumor, post-resection five-year survival rate for esophageal cancer is still lower than 30%. Multiple factors are found to influence the long-term results after surgical treatment. However, recent investigations have focused on the significance of lymph node matastasis (LNM), which seems to be one of the most important factors leading to poor survival. Hence, the prognostic significance of LNM in surgical resection of esophageal cancer was studied.

METHODS The rate and degree of LNM were evaluated and their prognostic significance was investigated through a retrospective study of 474 patients with esophageal cancer treated by surgery alone.

RESULTS LNM was positive in 211 patients, with an incidence of 44.5% (211/474). A total of 5382 lymph nodes were resected and studied pathologically, among which metastasis was found in 690 nodes with an overall LNM degree of 12.8% (690/5382). The 5-year survival rate was 30.6% (145/474) in the entire series, 12.8% (27/211) in patients with LNM, and 44.9% (118/263) in those without LNM.

CONCLUSIONS Surgery remains the first choice of treatment for carcinoma of the esophagus, and that meticulous lymph node dissection is an important practice of surgical oncology. However, in more advanced cases of this disease, surgery alone is of limited value in eradicating all cancer compromized tissue, and therefore the routine practice of extensive lymph node dissection in such cases may not be rewarding.

     

Over-expression of hypoxia-inducible factor 1 alpha increases angiogenesis of LNCaP cells

Objective:To evaluate the effect of HIF-1 α over-expression on angiogenesis in human prostate cancer cells. Methods:LNCaP cells(a human prostate cancer cell line) were transfected with the recombinant plasmid pcDNA3.1(-)-HIF-1α with Lipofectamine 2000 system. The positive clones were selected by G418 being further confirmed by Western blot and immunofluorescence. The expression levels of VEGF, iNOS and Ang- Ⅱ were determined. Results:The expression of HIF-1α in the LNCaP/HIF1α cells was significantly increased in transfected cells, which induced the up-regulation of VEGF, iNOS, whereas Ang- Ⅱ expression remained un- changed. Conclusion :Over-expression of HIF-1α can induce angiogenesis proteins and may improve the angiogenesis potency of prostate cancer.     

Acceleration of apoptosis by transfection of bak gene in multi-drug resistant (MDR) bladder cancer cellsI

Objective To observe the effects of bak gene on killing MDR bladder cancer cells and to study its mechanisms. Methods Bak gene was transfected into MDR bladder cancer cells by liposome. The mRNA of bak and bcl-2 were detected by in situ hybridization. The protein of bak and bcl-2 were detected by SABC immunohistochemistry. The growth rate of human bladder cancer cells was studied by constructing the growth curve, cell apoptosis being observed by flow cytometry, and the outline of cells observed by fluorescence stain. Results The expression of bak mRNA was positive in EJ/bak cells (64% ,P<0.05).Bak protein expression of EJ/bak cells was positive (60 % ) and bcl-2 protein expression was de creased (P<0.05). The growth of MDR bladder cancer cells was significantly inhibited by 32% after bak gene was transfected (P < 0. 05 ). Apoptosis cells increased significantly. The apoptosis rate was 35 %. Apoptotic bodies can be found in these cells on fluorescence stain. Conclusion Bak gene could inhibit the growth o     

Prognostic significance of MDR-1 P-glycoprotein expression in breast cancer

Objective: To investigate the expression of MDR-1 P-glycoprotein(MDR-1 Pgp) in breast cancer and analyze its correlation to the biological behavior and prognosis of the disease. Methods:The expression of MDR-1 Pgp was examined in 75 cases of breast cancer patients by using three different monoclonal antibodies(JSB1, C219 and C494) with S-P immunohistochemisty. These patients were followed up for 5 years, and the correlation between MDR-1 Pgp expression, survival rate and lymph metastasis was analyzed. Results: Positive detection of MDR-1 Pgp by JSB1, C219 and C494 in 75 cases of breast cancer was 86.7%, 48% and 85.3%, respectively. MDR-1 Pgp expression was not related to ages of patients (P > 0.05). JSB1-detected expression of MDR-1 Pgp was related to lymph node metastasis(P < 0.05); while C219 and C494 were not(P > 0.05). The patients with MDR-1 Pgp expression positively detected by either two of the three antibodies, had five-year survival rate that was significantly higher than those positively detected by all the three antibodies(P < 0.05). Conclusion:Three antibodies should be used simultaneously to detect MDR-1 Pgp expression in breast cancer. Positive MDR-1 Pgp expression in breast cancer detected by all the three antibodies may represent a poor prognosis; while positive MDR-1 Pgp detection by JSB1 and C494 is associated with lymph metastasis.     

Diagnostic significance of gastric cancer associated antigens (MG-AGS) in serum, ascitic fluid and gastric juice.

A group of monoclonal antibodies against gastric cancer, pooled in equal proportions, was used to investigate their corresponding antigens (MG-Ags) in serum and body fluid of patients with gastrointestinal cancer and benign diseases using microsphere-ELISA method. The mean serum level (plus 3 standard deviations) in 59 normal subjects was arbitrarily set as the positive threshold value. The positive rate was found to be 68.8% (135/196) in sera of patients with gastric cancer, 70% (14/20) in colonic cancer, 72.2% (24/33) in rectal cancer, 43.8% (7/16) in esophageal cancer, 45.5% (5/11) in cholecystic cancer and 34.9% (15/43) in lung cancer, which, however, was not found in primary liver cancer, pancreatic cancer and ovarian cancer. In 214 patients with benign diseases, a false positive rate was 7.48%. In gastric juice and ascitic fluid of patients with gastric cancer, the positive rates were found to be 61.7% (27/44) and 83.3% (20/24) respectively. These antigens were also determined repeatedly in sera of patients with gastric cancer who had undergone gastrectomy. It was found that the level of MG-Ags in sera began to decrease at 8-10 days after operation. These results suggest that the determination of MG-Ags is useful in the diagnosis of gastrointestinal cancer and evaluation of the treatments.

     

Expression of X-linked Inhibitor of Apoptosis Protein and Its Effect on Chemotherapeutic Sensitivity of Bladder Carcinoma

The expression of X-linked inhibitor of apoptosis protein (XIAP) gene and its effect on chemotherapeutic sensitivity in bladder carcinoma was explored. By using immunohistochemistry, the expression of XIAP was detected in 47 bladder carcinomas and 5 normal bladder tissues. The XIAP gene was transfected into bladder cancer cell line T24 by liposome and the positive clone was screened by G418. Cellular XIAP mRNA level was detected by RT-PCR. Low-dose mitocycin C was administered to induce the apoptosis of T24 cells. The in vitro growth of bladder carcinoma cells was analyzed by MTT colorimetry, and the apoptosis rate was assayed by TUNEL methods. It was found XIAP was moderately expressed in bladder carcinomas with the the positive rate being 78.73% (37/47), but the positive rate was not correlated with carcinoma stages and grades (P<0.05). XIAP mRNA level in transfected T24 cells was significantly increased by 3.8 times as compared with that in the cells not transfected with XIAP. After treatment with low-dose mitomycin C (0.005 and 0.05 mg/mL), the growth rate in XIAP no-transfected control group was increased by (11.60±0.25)% and (16.51±0.87)% (P<0.05), and the apoptosis rate was decreased by (10.1±0.2)% and (11.9±0.2%) (P<0.05) respectively as compared with XIAP transfected group. It was concluded that XIAP was expressed in most of bladder carcimoma samples. Overexpression of XIAP in T24 could significantly reduce the MMC-induced apoptosis of bladder carcinoma, suggesting its effect on the chemothera- peutic sensitivity of T24 cells.     

Establishment of an Infected Necrotizing Pancreatitis Model by Retrograde Pancreatic Duct Injection of Sodium Taurocholate and E. coli in Rats

A stable and reliable infected necrotizing pancreatitis （INP） model in rats was established in order to study the pathophysiological mechanism and pathological development role of INP and explore the new therapeutic methods for the diseases. Forty-six SD rats were randomly divided into 5 groups. The animals in group A received the injection of 5% sodium taurocholate into the pancreatic duct and those in group B underwent that of E. coli into the pancreatic duct. The rats in groups C, D and E were subjected to the injection of 5% sodium taurocholate in combination with different concentrations of E. coli （10^3, 10^4, 10^5/mE, respectively） into the pancreatic duct. The dose of injection was 0.1 mL/100 g and the velocity of injection was 0.2 mL/min in all the 5 groups. Eight h after the injection, the survival rate of animals was recorded and the surviving rats were killed to determine the serum content of amylase and perform pathological examination and germ cultivation of the pancreatic tissue. The results showed that acute necrotizing pancreatitis model was induced by injection of 5% sodium taurocholate into the pancreatic duct. The positive rate of germ cultivation in group A was 12.5%. The acute necrotizing pancreatitis model was not induced by injection of E. coli into the pancreatic duct and the positive rate of germ cultivation in group B was 0. The INP model was established in groups C to E. The positive rate of germ cultivation was 60%, 100% and 100% and 8-h survival-rate 100%, 100% and 70% in groups C, D and E, respectively. It was concluded that a stable and reliable model of INP was established by injection of 5% sodium taurocholate in combination with 10^4/mL E. coli into the pancreatic duct with a dose of 0.1 mL/100 g and a velocity of 0.2 mL/min. The pathogenesis of INP might be that the hemorrhage and necrosis of pancreatic tissue induced by sodium taurocholate results in weakness of pancreatic tissue in fighting against the germs. Meanwhile, the necrotic pancreatic tissue provides a good p     

Expression of p57kip2 and cyclinE proteins in human pancreatic cancer

Objective To investigate the effects of p57kip2 and cyclinE proteins on the genesis and progression of human pancreatic cancer. Methods The expression of p57kip2 and cyclinE proteins in tumor tissues and adjacent tissues of pancreatic cancer in 32 patients was detected by SP immunohistochemical technique. Results The p57kip2 protein positive-expression rate in tumor tissues of pancreatic cancer was 46.9%, which was lower than that in adjacent pancreatic tissue (P&lt;0.05). The p57kip2 protein positive-expression correlated significantly with tumor cell differentiation (P&lt;0.05) and did not correlate significantly with lymph node metastasis (P&gt;0.05). The cyclinE positive-expression rate in tumor tissues was 68.8%, which was higher than that in adjacent pancreatic tissues (P&lt;0.05). The cyclinE positive-expression also correlated significantly with tumor cell differentiation and lymph node metastasis (P&lt;0.05). The cyclinE protein positive-expression rate in the tumor tissues of the p57kip2 protein positive-expression group was lower than that in the p57kip2 protein negative-expression group, and there were no significant correlation between the two groups (r= -0.112, P＞0.05).Conclusion Decreased expression of the p57kip2 protein and/or over-expression of the cyclinE protein may play an important role in the genesis and progression of human pancreatic cancer.     

Hypoxia-Inducible Factor-lalpha Suppressing Apoptosis and Increasing Tolerance of Lung Cancer Cells to Chemotherapy

In order to construct plasmid of hypoxia-inducible factor-lalpha （HIF-1α）, and transfect into human lung cancer cells A549, the change in sensitivity of lung cancer cells A549 to chemotherapy was observed. HIF-1α mRNA structure region was amplified by RT-PCR and inserted into plasmid pcDNA3. The expression plasmid pcDNA3/HIF-1α was transfected into A549 with LipofectAMINE^TM2000. The expression of HIF-1α protein was detected by Western blot. After A549 cells were transfected with HIF-1α prior to addition of 5-Fu, the growth activity was measured by growth curve, apoptosis was detected by flow cytometry at 48 h, and the levels of caspase3 and MDR-1 were determined by Western blot. The results showed that the constructed expression plasmid was analyzed with restriction enzymes and gel electrophoresis. Two DNA lanes at 2.55 kb and 5.4 kb respectively were found, which were consistent with that expected. The growth rate in 5-Fu group was significantly inhibited, and the apoptosis index and caspase3 activity were increased significantly as compared with control group. After HIF-1α being transfected into A549, the activity of MDR-1 was increased and the effect of 5-Fu was weakened. In conclusion, HIF-1α can promote chemoresistance by increasing the activation of MDR1 and suppressing apoptosis during lung cancer cells A549 in- duced with 5-Fu.     

The expression of midkine (MK) in pancreatic carcinoma and its clinical significance

Objective To investigate the expression of midkine (MK) and its relation with angiogenesis, biological features and prognosis of pancreatic carcinoma (PC). Methods MK expression and microvessel density (MVD) were determined in 52 cases of human PC with immunohistochemistry and results were compared with pathology. Results Mean MVD of PC was 64 ± 18 and positive expression of MK was detected in 38 cases (73%). The positive rate of MK was significantly lower in cases of without metastasis and at early clinical stage (stage Ⅰ - Ⅱ) than that with metastasis and at stage Ⅲ - Ⅳ. MVD was significantly higher in MK-positive PC than in MK-negative PC (P < 0. 01). Follow-up showed that postoperative survival time was shorter in patients with positive expression of MK. Conclusion MK is closely related to angiogenesis PC. MK expression is one of the predictors for the biological behavior of PC. 4 refs, 1fig.