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Experimental Study of the Relationship between Nerve Growth Factor Expressions and Metastasis in Hepatocellular Cacinoma

Author: HeJun
Tutor: WangYuHua
School: Hebei Medical University
Course: Oncology
Keywords: Nerve Growth Factor Vascular endothelial growth factor E - cadherin Hepatocellular carcinoma Tumor recurrence Tumor metastasis Immunohistochemistry Flow cytometry
CLC: R735.7
Type: Master's thesis
Year: 2011
Downloads: 21
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Abstract


Objective: To detect the expression of different liver tissue and different metastatic potential human hepatoma cell line of nerve growth factor (nerve growth factor, NGF), vascular endothelial growth factor (vascular endothelial growth factor, VEGF), E-cadherin ( expression of E-cadherin, E-CD) in different liver tissue, to discuss the relationship between the NGF, VEGF, E-CD and hepatocellular carcinoma (human hepatocellular carcinoma HCC, HCC) in patients with clinical and pathological features relations and overall survival, seeking at the molecular level of the transfer of early diagnosis and treatment of liver cancer recurrence newer and more effective method. Methods: Using immunohistochemistry SP staining was detected of NGF, VEGF, E-CD cirrhotic liver tissue in 45 cases of liver cancer tissue and corresponding paraneoplastic, as well as the expression of the adjacent liver tissue in 15 cases of hepatic hemangioma (control group). NGF protein was measured by flow cytometry (flow cytometry, FCM) expression in SMMC-7721, HepG2 cell lines. Patients were followed up for overall survival time. Summarize the experimental results with liver cancer in patients with clinical and pathological data (gender, age, tumor size, AFP, HBsAg, HBeAb, clinical stage, histological grade, invasive grouping) and survival to explore the relationship between the protein and its overall survival time between the relationship with clinical and pathological data of patients with hepatocellular carcinoma and patients. Results: 1 hepatoma tissues of NGF, expression levels of VEGF, E-CD with paraneoplastic hardening of liver tissue and liver hemangioma liver tissue adjacent to compare: immunohistochemistry results showed that NGF, VEGF, E-CD positive for both groups liver tissue expression, cytoplasmic membrane yellow dye. NGF in hepatocellular positive expression rate was 68.9% (31/45), significantly higher than in adjacent cirrhotic liver tissue positive expression rate of 33.3% (15/45) (Z = -4.249, P = 0.000 lt; 0.05) and liver tissue adjacent hepatic hemangioma positive expression rate of 13.3% (2/15) (Z = -3.889, P = 0.000 lt; 0.05), next to the hardening of the liver tissue and liver hemangioma paracancerous compared to liver tissue showed no significant difference (Z = -1.586, P = 0.113 gt; 0.05). VEGF expression in HCC positive rate of 77.8% (35/45), significantly higher than in adjacent cirrhotic liver tissue positive expression rate of 37.8% (17/45) (Z = -5.297, P = 0.000 lt; 0.05) and liver tissue adjacent hepatic hemangioma positive expression rate of 60.0% (9/15) (Z = -3.281, P = 0.000 lt; 0.05), adjacent noncancerous liver tissue adjacent to the hardening of the liver tissue and liver hemangioma was no significant difference compared to (Z = -1.297, P = 0.194 gt; 0.05). The positive expression of E-CD in HCC was 40.0% (18/45), and its expression was significantly positive expression rate of hardening of the liver tissue than in adjacent 88.9% (40/45) (Z = -5.504, P = 0.000 lt ; 0.05) liver tissue adjacent hepatic hemangioma positive expression rate of 93.3% (14/15) (Z = -4.753, P = 0.000 lt; 0.05), next to the hardening of the liver tissue and liver hemangioma paracancerous liver tissue showed no statistically significant (Z = -1.305, P = 0.192 gt; 0.05). 2 HCC tissues of NGF, the relationship between the expression levels of VEGF, E-CD: Statistical analysis showed that liver cancer tissues, the expression levels of NGF and VEGF expression levels were the rating positively correlated (rs = 0.482, P = 0.001 lt; 0.05 ), the level of expression of of NGF the expression level and the E-CD was Level negative correlation (rs = -0.467, P = 0.001 LT; 0.05), the expression level of the level of the expression of VEGF and the E-CD's was Level negative correlation (rs = - 0.509, P = 0.000 lt; 0.05). 3 HCC tissues of NGF, VEGF, E-CD expression levels in liver cancer patients with clinical pathological features of the relationship between: Statistical results show that: NGF in later clinical stage, poorly differentiated, high invasive group hepatocellular carcinoma tissues than clinical earlier stages (χ to 2 = 11.463, P = 0.022 lt; 0.05), well-differentiated the (χ ~~ 2 = 12.730, P = 0.002 lt; 0.05), low invasive group (Z = -2.267, P = 0.023 lt; 0.05), and liver cancer in patients with clinical stage (rs = 0.444, P = 0.002 lt; 0.05), pathological grade (rs = 0.529, P = 0.000 lt; 0.05), the risk of recurrence and metastasis after (rs = 0.342, P = 0.022 lt; 0.05) showed a level of positive correlation; VEGF in clinical staging late, poorly differentiated, the high invasion group liver cancer tissues than clinical stage earlier (χ to 2 = 14.858, P = 0.005 lt; 0.05), well-differentiated (χ to 2 = 6.579, P = 0.037 lt; 0.05), low-invasive group (Z = -1.987, P = 0.047 lt; 0.05) by and with liver cancer in patients with clinical stage (rs = 0.570, P = 0.000 lt; 0.05), pathological grade (rs = 0.366, P = 0.011 lt; 0.05), the risk of recurrence and metastasis (rs = 0.300, P = 0.046 lt; positive correlation of 0.05) were grade; E-CD in clinical stage early, well-differentiated the low invasive group hepatocellular carcinoma tissues than later clinical stage (χ to 2 = 9.577, P = 0.048 lt; 0.05), poorly differentiated (of χ ~~ 2 = 23.139, P = the 0.000 lt; 0.05), high invasiveness group (Z = -2.134, P = 0.033 lt; 0.05) by its expression and liver cancer in patients with clinical stage (rs = -0.382, P = 0.010 lt; 0.05), pathological grade (rs = -0.656, P = 0.000 lt; 0.05), the risk of recurrence and metastasis (rs = -0.322, P = 0.031 lt; 0.05) was level negative related; NGF, VEGF, E-CD expression in hepatocellular carcinoma tissue with the patient's gender, age, tumor diameter, AFP, HBsAg, HBeAb unrelated (P gt; 0.05). NGF expression level comparison: Flow cytometry showed NGF protein expression in highly metastatic human hepatoma cell line SMMC-7721 fluorescence index (FI) was 0.609 ± 0.071, higher than in non-metastatic potential in 4 different metastatic potential cell lines expression of human hepatoma cell line HepG2 fluorescence index 0.537 ± 0.092, both compared to the differences were statistically significant (P = 0.030 lt; 0.05). 5 survival analysis: the whole group of 45 patients were followed up for 1.5 to 31.0 months, with an average survival time of 22.1 ± 1.6 months, the overall survival of more than 1 year in 32 cases, the 1-year survival rate was 71.1% (32/45) . NGF expression in HCC tissues (-) and (~) by overall survival rates were 78.6% (11/14) and 45.2% (14/31), the average survival time was 28.1 ± 1.0 months and 19.1 ± 2.0 months ( P lt; 0.05); VEGF expression in HCC tissues (-) and (~) the total survival rates were 90.0% (9/10) and 45.7% (16/35), with an average survival time was 30.3 ± 0.7 months E-CD and 19.8 ± 1.9 months (P lt; 0.05); expressed in HCC tissues (-) and (~) by overall survival rates were 37.0% (10/27) and 83.3% (15/18), with an average The survival time was 17.7 ± 2.1 months and 28.8 ± 1.2 (P lt; 0.01). Univariate analysis showed that the patient's age, gender, tumor diameter, HBsAg yin and yang, HBeAb yin and yang NGF, the expression of VEGF-negative, E-CD expression positive survival (Log rank test, P lt; 0, 05); impact on overall survival time of patients with hepatocellular carcinoma difference was not statistically significant (P gt; 0.05), clinical stage earlier pathological differentiated, serum AFP lt; 400μg / L, low invasive group compared with clinical staging than night, pathological differentiation poor serum AFP ≥ 400μg / L, higher survival rates of invasive group (Log rank test, P lt; 0.05). Multivariate COX risk model analysis results show that only NGF (RR = 7.797, P = 0.006 lt; 0.05), VEGF (RR = 8.108, P = 0.042 lt; 0.05) expression levels and tumor aggressiveness (RR = 5.434, P = 0.003 lt; 0.05) is an independent factor affecting survival of liver cancer patients. Conclusion: NGF expression of VEGF in HCC tissues than paracancerous next to the hardening of the liver tissue and liver hemangioma liver tissue was significantly elevated, later clinical stage, differentiation is worse, aggressive and higher of NGF of VEGF positive expression Vietnam overall survival time is short, the worse the prognosis. 2 E-CD expression in hepatocellular carcinoma and adjacent noncancerous next to the hardening of the liver tissue and liver hemangioma liver tissue was significantly reduced, later clinical stage, differentiation is worse, invasive higher the positive expression of E-CD downregulated or missing, the shorter overall survival time, the worse the prognosis. 3 clinical staging, pathological grade, invasive and AFP levels of overall survival time of patients with hepatocellular carcinoma differences, clinical stage earlier pathological differentiated, AFP lower and are low in patients with invasive group, survival higher and longer overall survival, the prognosis is good. 4 HCC tissue expression of NGF and metastasis of HCC indicators of VEGF expression was level positive correlation was with another transfer Index E-CD expression level negatively correlated, suggesting that NGF in the incidence of liver cancer, plays a vital role in the development with liver cancer recurrence and metastasis of malignant behavior is closely related to NGF with clinical and pathological data jointly can effectively predict the risk of recurrence and metastasis of liver cancer, the risk of recurrence and metastasis of liver cancer patients to take effective preventive interventions to reduce or prevent surgery clinical recurrence and metastasis after providing a theoretical basis, is expected to become a target for liver cancer treatment and assessment of prognosis.

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