血管内皮生长因子-C在肾癌中的表达及意义

目的:探讨血管内皮生长因子-C(VEGF-C)在肾癌发生发展中的作用。方法:应用免疫组织化学SP法检测VEGF-C在肾癌及正常肾组织中的表达情况。结果:肾癌组织VEGF-C的阳性表达率(85.0%)显著高于正常肾组织(P<0.01)。肾癌组织VEGF-C的阳性表达与年龄、性别、分化程度无关(P>0.05),与有无淋巴结转移和肾癌临床分期有关(P<0.05)。结论:VEGF-C可能通过促进淋巴管生成参与肾癌的淋巴结转移,在肾癌的发生发展中发挥重要作用。

Vascular Endothelial Growth Factor C Expression in Gastric Carcinoma and Its Relationship with Lymph Node Metastasis

Objective: To study the expression of vascular endothelial growth factor C (VEGF-C) in gastric carcinoma and its relationship with lymph node metastasis. Methods: The expression of VEGF-C mRNA in 5 gastric carcinoma cell lines was detected by reverse transcription-polymerase chain reaction (RT-PCR). Simultaneously, the expression of VEGF-C protein in gastric carcinoma tissues, which were obtained from 63 patients who underwent radical gastrectomy, was detected by immunohistochemistry. Results: Three of the 5 gastric carcinoma cell lines, MKN-45, SGC-7901 and AGS, expressed VEGF-C mRNA. VEGF-C protein was expressed in 52.4% (33/63) of patients. VEGF-C protein expression was more frequently found in tumors with lymph node metastasis than in those without (P<0.01). VEGF-C protein expression was also closely related to lymphatic invasion (P<0.01) and TNM stage (P<0.01). However, there was no significant correlation between VEGF-C expression and the age, gender, tumor size, tumor location, Lauren classification, depth of invasion, and vascular invasion. Conclusion: The expression of VEGF-C is closely related to lymph node metastasis of gastric carcinoma, and lymphangiogenesis might be a new target for treatment of gastric carcinoma.

Expression of vascular endothelial growth factor-C and angiogenesis in esophageal squamous cell carcinoma

AIM： To investigate the expression of vascular endothelial growth factor-c （VEGF-C） mRNA and microvessel density （MVD） in human esophageal squamous cell carcinoma （ESCC） and its relationship with clinical significance. METHODS： Specimens obtained from 43 patients undergoing surgical resection for ESCC were used in this study. The expression of VEGF-C mRNA was examined by in situ hybridization. Tumor MVD was determined immunohistochemically with anti-CD31 antibody and estimated by image analysis. Ten sections of adjacent normal mucosa were also examined. RESULTS： VEGF-C mRNA expression was detected in cytoplasm of carcinoma cells. Of the 43 ESCC patients studied, 18 cases （41.9%） were positive for VEGF-C mRNA. No VEGF-C mRNA expression was observed in normal esophageal mucosa. VEGF-C mRNA expression correlated significantly with lymph node metastasis, TNM stage and depth of invasion （P 〈 0.05）. Furthermore, histological grade （differentiation） tended to correlate with VEGF-C mRNA expression, but was not statistically significant （P 〉 0.05）. In tumor lesions, the MVD was significantly greater than that in normal esophageal mucosa. MVD correlated significantly with lymph node metastasis, TNM stage and depth of invasion （P 〈 0.05）, but not with histological grade （differentiation） （P 〉 0.05）. Lesions with VEGF-C mRNA expression had a significantly higher MVD than that of those without VEGF-C mRNA expression （P 〈 0.05）. CONCLUSION： VEGF-C plays a role in lymphatic metastasis via lymphangiogenesis and angiogenesis in ESCC. VEGF-C is one of the important predictors of the biological behavior in ESCC.

Association of serum vascular endothelial growth factor-C and lymphatic vessel density with lymph node metastasis and prognosis of patients with gastric cancer

AIM： To investigate whether serum vascular endothelial growth factor-C （SVEGF-C）, VEGF-C, and lymphatic vessel density （LVD） in tumor tissues are related to lymph node metastasis （LNM） and prognosis in gastric cancer. METHODS： SVEGF-C levels of 80 gastric cancer patients and 20 healthy donors were examined using ELISA. VEGF-C expression and LVD were examined using immunohistochemical staining. Kaplan-Meier survival analysis was performed to determine their influence on the prognosis of the patients. RESULTS： The SVEGF-C level in gastric cancer patients （595.9 ± 201.0 ng/L） was significantly higher （P = 0.000） than controls （360.0 ± 97.4 ng/L）. Both SVEGF-C and LVD were significantly higher in poorly differentiated adenocarcinomas, T3 and T4, LNM, distant metastasis, and pTNM groups Ⅲ and Ⅳ （P = 0.000）. The sensitivity and specificity of SVEGF-C for predicting LNM were 82.8% and 81.8%, respectively （cut-off = 542.5 ng/L）. The positive expression rate of VEGF-C was significantly higher in cancerous than in normal tissues （65% vs 20%; P = 0.001）. VEGF-C expression up-regulation was significantly related to differentiation, depth of invasion, LNM, distant metastasis, and pTNM stage （P = 0.000）. LVD was 10.7 ± 3.1/200 HP in the experimental group vs 4.9 ± 1.3/200 HP in controls （P = 0.000）; LVD in cancerous tissues with and without LNM was 12.0 ± 2.7/200 HP vs 7.6 ± 0.5/200 HP, respectively （P = 0.000）. SVEGF-C and LVD were significantly higher in VEGF-C positive than in negative patients （P = 0.000）; SVEGF-C level was related to LVD （P = 0.000）. Kaplan-Meier survival analysis factors predicating poor prognosis were： SVEGF-C level （P = 0.001）, VEGF-C expression and LVD （both P = 0.000）. CONCLUSION： SVEGF-C level, VEGF-C and LVD are related to LNI and poor prognosis of patients with gastric cancer, SVEGF-C may be a biomarker for LNI in gastric cancer,

Expression of vascular endothelial growth factors A and C in human pancreatic cancer

AIM： To study the expression of vascular endothelial growth factor A （VEGF-A） and VEGF-C and to determine whether the presence of VEGF-A and VEGF-C was associated with the clinicopathologic characteristics of pancreatic cancer. METHODS： VEGF-A and VEGF-C mRNA transcripts were examined by Northern blot in 6 human pancreatic cancer cell lines and 8 normal pancreatic tissues and 8 pancreatic carcinoma specimens. The expression of VEGF-A and VEGF-C proteins was examined by Western blot in the tested cell lines and by immunohistochemical stain in 50 pancreatic carcinoma samples. RESULTS： VEGF-A and VEGF-C mRNA transcripts were present in all the 6 human pancreatic cancer cell lines. Immunoblotting revealed the presence of VEGF-A and VEGF-C proteins in all the cell lines. Northern blot analysis of total RNA revealed 3.0-fold and 3.6-fold increase in VEGF-A and VEGF-C mRNA transcript in the cancer samples, respectively. Immunohistochemical analysis confirmed the expression of VEGF-A and VEGF-C in cancer cells within the tumor mass. Immunohistochemical analysis of 50 pancreatic cancer tissue samples revealed the presence of VEGF-A and VEGF-C immunoreactivity in 50% and 80% of the cancer tissue samples, respectively. The presence of VEGF-A in these cells was associated with larger tumor size and enhanced local spread （x^2= 6.690, P= 0.035〈0.05） but was not associated with decreased patient survival. However, the presence of VEGF-C in the cancer cells was associated with increased lymph node metastasis （x^2= 5.710, P= 0.017〈0.05）,but was not associated with decreased patient survival. There was no correlation between the expression of VEGF-A and VEGF-C in the same cancer cells. CONCLUSION： VEGF-A and VEGF-C are commonly overexpressed in human pancreatic cancer and may contribute to tumor growth and lymph node metastasis, There is no relationship between the expression of VEGF-A and VEGF-C in pancreatic cancer.

Relationship between the Expression of Vascular Endothelial Growth Factor C and Lymphangiogenesis in Human Pancreatic Cancer

Objective： To investigate the role the vascular endothelial growth factor C （VEGF-C） expression in lymphangiogenesis and lymph node metastasis in pancreatic cancer. Methods： The expression of VEGF-C in pancreatic cancer tissue of 42 cases and normal pancreatic tissue of 10 cases was detected by using immunohistochemical S-P method. The microlymphatic vessel density （MLVD） of both tissues was measured by enzyme-histochemical method at the same time. Results： The VEGF-C positive expression rate was significantly higher in pancreatic cancer tissue （64.3%） than in normal pancreatic tissue （20%, P〈0.05）. The MLVD was significantly higher in pancreatic cancer tissue （21.93±6.39） than that in normal pancreatic tissue （4.40±2.07, P〈0.001）. The VEGF-C positive expression rate, MLVD and lymph node metastasis had a positive correlation. Conclusion： The VEGF-C may play an important role in the lymphangiogenesis and lymph node metastasis of pancreatic cancer.

Altered expression of mitochondrial related genes in the native Tibetan placents by mitochondrial cDNA array analysis

Objective： To explore the mechanism of native Tibetan fetuses adaptation to hypoxia, we tried to find the different expression genes about mitochondrial function in the native Tibetan placents. Methods： In this study, the placents of native Tibetan and the high-altitude Hart （ha-Hart） were collected. After the total RNA extraction, the finally synthesized cDNAs were hybridized to mitochondrial array to find the altered expression genes between them. Then, the cytochrome c oxidase 17 （Coxl7）, dynactin 2 （DCTN2, also known as p50）, and vascular endothelial growth factor receptor （VEGFR, also known as KDR） were chosen from the altered expression genes to further verify the array results using the SYBR Green real-time PCR. Because the altered expression genes （such as Cybb and Cox 17） in the array results related to the activities of COXI and COXIV, the placental mitochondria activities of COXI and COXIV were measured to find their changes in the hypoxia. Results： By a standard of≥1.5 or ≤0.67, there were 24 different expressed genes between the native Tibetan and the ha-Han placents, including 3 up-regulated genes and 21 down-regulated genes. These genes were related to energy metabolism, signal transduction, cell proliferation, electron transport, cell adhesion, nucleotide-excision repair. The array results of Cox17, DCTN2 and KDR were further verified by the real-time RT-PCR. Through the mitochondria respiration measurements, the activity of COXI in the native Tibetan placents were higher than that of ha-Han, there was no difference in COXIV activity between them. Conclusion： The altered mitochondrial related genes in the native Tibetan placents may have a role in the high altitude adaptation for fetuses through changing the activity of mitochondrial COX.

Effects of carbon dioxide pneumoperitoneum on metastasis of ovarian carcinoma cell line SKOV_3

The long suspicion of the potential harm of carbon dioxide （CO2） pneumoperitoneum exists in laparoscopic cancer surgery. For better understanding of this problem, we targeted this study at the effects of CO2 pneumoperitoneum on the invasive ability of ovarian carcinoma cell line and the possible mechanism within it. To study the effects of CO2 pneumoperitoneum on carcinoma cell, SKOV3 cells were divided into 2 groups, respectively exposed to pneumoperitoneal CO2-insuffiation and normal conditions. To study the possible mechanism, SKOV3 cells were divided into 3 groups, one was exposed to CO2 pneumoperitoneum, one to N2 and the other to normal conditions served as control. The in vitro adhesive and invasive ability of the cells was analyzed through 3-（4,5-dimethylthiazol-2-yl）-2,5-diphenyl tetrazolium bromide （MTT） assay and Boyden filters metastasis model; the expressions of vascular endothelial growth factor C （VEGF-C） and matrix metalloproteinase 9 （MMP9） were determined by reverse transcription polymerase chain reaction（RT-PCR）, and Western blot. We found that the adhesive ratio of SKOV3 cells exposed to CO2 was significantly higher than that of the control group; cells exposed to CO2 invaded the matrigel with a greater number （P〈0.01）; the expression of VEGF-C exposed to both CO2 and N2 was significantly increased compared with control group （P〈0.05）; the MMP9 expression level of CO2 group was higher than that of N2 group, P〈0.05. We concluded that carbon dioxide pneumoperitoneum may improve the adhesive and invasive ability of ovarian carcinoma cell line in vitro and CO2 can also be an independent factor to stimulate the expression of MMP9.

Expression of Vascular Endothelial Growth Factor C and Its Clinical Significance in Human Esophageal Squamous Cell Carcinoma

OBJECTIVE To examine the expression of vascular endothelial growth factor C （VEGF-C） in human esophageal squamous cell carcinoma （ESCC）, and to clarify its role in lymphatic metastasis in ESCC patients.METHODS Esophageal carcinoma EC9706 cells and samples from 49 patients with primary ESCC were investigated by using S-P immunohistochemistry （IHC）, the semi-quantitative reverse transcriptase-polymerase chain reaction （RT-PCR） and in situ hybridization （ISH） methods for VEGF-C expression. RESULTS VEGF-C positive expression was found in EC9706 cells through IHC, ISH and RT-PCR. Positive IHC for VEGF-C was observed in 36 of 49 cases of ESCC. There was a significant difference between the expression of VEGF-C in a lymph-node-positive group compared to a node-negative group （χ^2=4.7, P〈0.05）. Positive ISH for VEGF-C mRNA was observed in 23 of 49 cases of ESCC. There was a significant difference between the expression of VEGF-C in the lymph-node-positive group and node-negative group （χ^2=31.3, P〈0.01）. The expression of VEGF-C was significantly higher in the lymph-node-positive group compared to the node-negative group. Of 49 ESCC tissues, RT-PCR for VEGF-C mRNA was observed positively in 29 cases. There was a significant difference between the expression of VEGF-C in the lymph-node-positive group and node-negative group （χ^2=23.3, P〈0.01）. The expression of VEGF-C was significantly higher in the lymphnode-positive group compared to the node-negative group. Expressions of VEGF-C were not significantly associated with age, gender, and pathological grade. There was a relationship between VEGF-C mRNA expressions by RT-PCR and ISH （χ^2=18.5, P〈0.01） in ESCC cases, but with no significant difference between the two methods. CONCLUSION VEGF-C expression may induce lymphangiogenesis in human ESCC. There was a close correlation between VEGF-C expression and lymph node metastasis. VEGF-C can serve as a useful prognostic factor for ESCC patients.

Correlation between the expression of vascular endothelial growth factor c and C-erbB-2 in human breast cancer

Objective: We aimed to study the transcription level of VEGF-C in human breast cancer tissue, and explore the correlations with the expression of C-erbB-2. Methods: The expression of VEGF-C mRNA in 51 cases of human breast cancer was assessed by hybridization in situ. The expressions of C-erbB-2 was assessed by immunohistochemistry. Results: The positive rate of VEGF-C mRNA was 54.9% in 51 cases of breast cancer. The transcription level had correlation with tumor size and status of lymph nodes(P < 0.05). The expression of VEGF-C mRNA had a positive correlation with the expression of C-erbB-2(P < 0.05). Conclusion: The up-expression of VEGF-C has a significant correlation with the malignancy level and clinical stage of breast cancer. The combined detection of VEGF-C, C-erbB-2 may help to estimate the prognosis of patients with breast cancer and study on thetherapeutic implications.

EphA2与VEGF-C在胃癌中的表达及其与淋巴管生成的关系

目的:探讨胃癌组织中EphA2与VEGF-C的表达及临床意义。方法:用免疫组化法检测82例胃癌组织及其癌旁组织中EphA2与VEGF-C蛋白的表达,以及淋巴管密度(LVD);用real-timePCR检测20例胃癌组织及其癌旁组织中EphA2与VEGF-Cm RNA的表达。分析EphA2与VEGF-C表达与患者临床病理特点及淋巴管生成的关系。结果:EphA2与VEGF-C在胃癌组织中的阳性表达率均明显高于癌旁正常组织(65.8%vs.42.6%;71.9%vs.39.0%,均P<0.05)。EphA2与VEGF-C的高表达与肿瘤浸润深度、淋巴结转移、TNM分期有关(均P<0.001)。EphA2蛋白与VEGF-C蛋白高表达的胃癌组织中LVD计数分别明显高于两者低表达胃癌组织(15.25±5.41vs.10.95±5.41,P=0.001;14.87±5.71vs.11.00±5.01,P=0.006)。胃癌组织中EphA2与VEGF-C蛋白以及m RNA表达均呈正相关(r=0.375,P=0.001;r=0.559,P=0.01)。结论:EphA2与VEGF-C在胃癌组织中均呈高表达,且可能共同促进淋巴管生成与胃癌淋巴结转移。

生存素、FHIT、VEGF-C在胃腺癌中的表达及相关性

目的探讨生存素(survivin)、脆性组氨酸三联体(FHIT)、血管内皮生长因子C(VEGF-C)在胃腺癌中的表达及其相关性。方法用半定量RT-PCR检测43例胃腺癌及对应癌旁、远癌正常组织中生存素、FHIT、VEGF-C mRNA的表达。结果胃腺癌组织中生存素、VEGF-C的mRNA表达水平明显高于癌旁和远癌正常组织,FHIT mRNA表达明显低于癌旁和远癌正常组织(P<0.05)。生存素、VEGF-C mRNA在胃腺癌中的表达均与肿瘤的分化、分期、浸润、淋巴结转移有关(P<0.05);FHIT mRNA在胃腺癌中的表达与肿瘤直径、分期、浸润、淋巴结转移有关(P<0.05)。胃腺癌中生存素与VEGF-C的mRNA表达呈正相关(P<0.05),生存素与FHIT的mRNA表达呈负相关(P<0.05)。结论生存素、VEGF-C表达增强与FHIT表达缺失共同促进胃癌的发生、发展。

Efficacy and effect on serum VEGF-C of mild moxibustion plus functional exercise for upper-limb lymphedema after breast cancer surgery

Objective To observe the efficacy of mild moxibustion combined with functional exercise in the treatment of upper-limb lymphedema after breast cancer surgery and its effect on serum vascular endothelial growth factor C(VEGF-C).Methods Seventy-eight patients were divided into a control group and an observation group by the random number table method,with 39 cases in each group.The control group received functional exercise and the observation group received mild moxibustion plus functional exercise.The differences in circumference between the two upper limbs,the lymphatic flow of the affected upper limb,the disability of arm,shoulder and hand(DASH)score,the functional assessment of cancer therapy-breast(FACT-B)score and serum VEGF-C level between the two groups were compared before and after treatment.Efficacy was evaluated after treatment.Results The total effective rate was significantly higher in the observation group than in the control group(P<0.05).The difference in circumference between the two upper limbs of the two groups decreased significantly after treatment(both P<0.05),but it was significantly lower in the observation group than in the control group(P<0.05).The lymphatic flow of the affected upper limb of the two groups increased significantly after treatment(both P<0.05),but it was significantly greater in the observation group than in the control group(P<0.05).The DASH scores of the two groups decreased significantly after treatment(both P<0.05),but it was significantly lower in the observation group than in the control group(P<0.05).The FACT-B scores of the two groups increased significantly after treatment(both P<0.05),but it was significantly higher in the observation group than in the control group(P<0.05).After treatment,the serum VEGF-C level increased significantly in the observation group(P<0.05),whereas the control group did not show significant change(P>0.05).The post-treatment serum VEGF-C level was significantly higher in the observation group than in the control group(P<0.05).Conclusion The efficacy of mild moxibustion combined with functional exercise for upper-limb lymphedema after breast cancer surgery is certain,which can reduce the difference in circumference between the two upper limbs,increase the lymphatic flow of the affected upper limb,improve the limb function and the quality of life,and regulate the serum VEGF-C level.

Isoflurane induces expression of vascular endothelial growth factor through activating protein kinase C in myocardial cells

Objective： Vascular endothelial growth factor （VEGF） plays important roles in establishing collateral circulation of ischemic myocardium. This study aimed to investigate the effect of isoflurane on VEGF expression and the potential intracellular signal transduction pathway in cultured rat myocardial cells in order to further reveal the molecular mechanism of myocardial preservation of isoflurane. Methods： Primary myocardial cells of Sprague-Dawley rats were isolated and cultured. They were divided randomly into control group, isoflurane group, protein kinase C （PKC） inhibitor group and PKC inhibitor＋isoflurane group where cells were respectively incubated without any treatment, treated by 0.5, 1.0 and 1.5 minimum alveolar concentration （MAC） of isoflurane for 6 hours, by PKC inhibitor calphostin C at a final concentration of 50 nmol/L and by 50 nmol/L calphosfin C＋ 1.0 MAC isoflurane for 6 hours. VEGF expression was detected by enzyme-linked immunosorbent assay （ELISA） and the expression levels of PKC isoforms were determined by Western immunoblotting method. Results： Isoflurane increased the VEGF expression in myocardial cells in a dose-dependent way. VEGF levels were significantly higher in 1.0 and 1.5 MAC isoflurane groups than in the control group （both P〈0.01）. The effect of isoflurane on upregulating VEGF expression was blocked by PKC inhibitor calphostin C （P〈0.01）, but calphostin C did not alter VEGF expression （P〉0.05）. Isoflurane induced the activation and translocation of PKC Immunoblotting analysis revealed that the immunoreactivity of PKC ε increased significantly in the membrane fractions and deceased significantly in the kytoplasm fractions for cells treated with 1.0 MAC isoflurane as compared with the untreated cells, but not of PKC a, PKCα and PKCζ （P〈0.01）. Conclusion： Isoflurane induces myocardial cells to release VEGF through activating PKCε from the endochylema to the cytomembrane, suggesting a possible novel mechanism of isoflurane protecting myocardial cells.