| Purpose:To explore the pathomechanism in carcinogenesis and development of advanced pancreatic cancer. To study mechanism of 125I seed in treating advanced pancreatic cancer and the relationgship between Nerve growth factor (NGF) and treatmeat effect.Method:1.58 cases with pancreatic cancer who received surgery between January 2011 to June 2014 in the first people’s hospital of Yunnan province were enrolled. Tumor tissue cut partly during surgery was set as experiment sample and Tumor tissues were excised as experimental specimens, and tissues adjacent to cancer were set as control. Among all the cases,39 were male,19 were female, and age 38-72 (57.14±8.82).14 cases were stage Ⅱ (24.14%),17 cases were stage Ⅲ (29.31%),27 cases were stage Ⅳ (46.55%) according to TNM Standard Tumor Staging. The average size of tumor was 22.15±11.04mm。There were 17 cases (29.31%) were well differentiated,19 cases (32.76%) were moderately differentiated and 22 cases (37.93%) were poorly differentiated after post-surgery pathology. Meanwhile,38 cases (65.52%) were confirmed lymphaden diversion, and the rest 20 cases (34.48%) without lymphaden diversion. Immunohistochemistry and Western blot were used to determine expression of nerve growth factor (NGF) and precursor of nerve growth factor (ProNGF) in tissue of advanced pancreatic cancer and tissues adjacent to cancer. Gel-Pro analyzer 4 image analysis system was used to analyse density of band. All the tests were repeated three times then analysed with the three IOD data by statistics.2.54 cases with advanced pancreatic cancer who were dignosed and treated with 125I seed implantation in our hospital from January 2011 to May 2014 were set as pancreatic carcinoma group (PCa group), the other 50 healthy volunteers were set as control (HC group). Enzymelinked Immunosorbent Assay (ELISA) was used to test serum NGF level and compared with corresponding tissue NGF level.3. CT was used to measure the shortest diameters before operation and 6th after operation. RECIST standard was applied to evaluate treatment effect. Complete remission, partial remission and stabilization were thought as effective, otherwise process aggravation was ineffective. Analyse the relation between serum NGF and treating effect of 125I seed and make receiver operating characteristic curve of srrum NGF to judge its diagnositic ability in advanced pancreatic cancer treated by 125I seed implantation.4.To set up pancreatic cancer transplantation tumor model of athymic mouse. Restraining tumor rate of 125I seed implantation treatment was calculated. Serum and tissue NGF level before and after treated were tested by ELISA and their relationship was analysed by statistics.Results:1. Immunohistochemical showed expression of NGF in tumor tissue powerfully positive, which displayed brown matter in cytoplasm increased obviously. Expression of NGF in tissues adjacent to cancer was lower, which displayed brown matter in cytoplasm was lower than that in tumor tissue. ProNGF dyeing showed brown matter in cytoplasm of tissues adjacent to cancer was more, meanwhile brown matter in cytoplasm of tumor tissues was less, which indicates expression of proNGF in tumor tissue was lower than in issues adjacent to cancer. Cell counting showed NGF positive cell population in tumor tissue was++++, and in tissues adjacent to cancer was++. ProNGF positive cell population in tumor tissue was+, and in tissues adjacent to cancer was++++. NGF positive cell was more in tumor tissues than that in tissues adjacent to cancer, and proNGF positive cell was more in tissues adjacent to cancer than in tumor tissues, which presented statistically significant difference by rank sum test (p<0.001). Western blot analysis showed expression of NGF protein in tumor tissues higher than that in issues adjacent to cancer, but expression of proNGF in issues adjacent to cancer was higher than that in tumor tissues. Semiquantitative western showed, IOD value in tumor tissues and tissues adjacent to cancer were 245.00±7.55 and 86.33±7.37, respectively, which showed statistical significance (P=0.002).2.6 cases of total 54 cases in PCa group were exclused for death within 3 months after 125I seed implantation and other 3 cases were also exclused for distant aversion. Finally,45 cases wre enrolled to study, among whom,30 were male and 15 were female, mean age 52.07±8.43 (41-72).32 cases were male and 18 cases were female, mean age 55.23±7.21 (42-75) in HC group. There were no significant differences in age and sex construction between PCa group and HC group (p>0.05). 27 cases were stage Ⅲ (60%),18 cases were stage IV (40%) according to TNM Standard Tumor Staging. ELISA showed NGF level in PCa group was mean 775.60± 250.97 pg/ml, and 35.03±25.36 pg/ml in HC group. Serum NGF level in advanced pancreatic carcinoma was higher than that in HC group with elevated extent about 22 time, which was significantly statistically different (p<0.0001). Tumor tissue NGF level of the same subject which was detected by ELISA too was 1848.61±1426.45 pg/ml in average. NGF level in serum and tumor tissue of the same patients which were analysed by Pearson correlation analysis showed positive correlation (r= 0.487, P=0.001).3.14 cases in all cases who received 125I seed implantation were considered as effective, among whom 9 cases (20.00%) were thought as partial remission, and 5 cases (11.11%) as stabilization. The rest 31 cases (68.89%) who were process aggravation was thought as ineffective. The mean serum NGF level in effective group was 537.42±122.61 pg/ml, meanwhile 883.17±217.79 pg/ml in ineffective group. NGF level in effective group is higher than that in effective group, which was significantly statistically different (p<0.05)。4. To analyse subjects’serum NGF level and treatment effect by ROC curve, the result showed area under the curve (AUC) of NGF was 0.945, standard deviation was 0.032,95% confidence interval was 0.882-1.00. The level of NGF was 649.59 pg/ml at the breakpoint in top-left of curve with sensitivity 90.30%, specificity 85.70%. There are 12 cases with effective treatment and 3 cases with ineffective among the 15 cases with serum NGF<649.59 pg/ml. There are 2 cases with effective treatment and 28 cases with ineffective among the 15 cases with serum NGF>649.59 pg/ml. Positive predictive value 80%(12/15) and negative predictive value 93.33%(28/30).4. The time during mouses erupted tumor which could be seen was 7-13d, and mean time (9.43±2.17) d. It took 26-38 d until the tumor growed to 1000 mm3, mean (32.21±3.62) d. Compared with baseline, volume of subcutaneously tumor in model control group was increased uninterruptedly at 4 week after surgery, which was statistically significant (P< 0.001). Meanwhile, volume in treatment group increased which wasn’t statistically significant (P>0.005), but became to decreased since the second week, which was statistically significant (P<0.005). The volumes of subcutaneously tumor in model control group and treatment group were 1009.80±39.21 mm3 and 1018.00±48.33 mm3 respectively, which was statistically significant (P> 0.005) and volumes in both groups at 4 week were 1582.60±54.91 mm3 和 781.80±86.32 mm3 respectively, which was statistically significant (P< 0.005).Conclusion:Expression of NGF increasing and proNGF decreasing could be one of the mechanism leading to pancreatic cancer carcinogenesis and development, invasion and migration. Serum NGF that had positive correlation with tissue NGF level, could forecast treatment effect of 125I seed implantation well. Thus, level of serum NGF level could be a reference index to predict the therapeutic efficacy of 125I seed implantation in advanced pancreatic cancer patients. |
PDF Full Text Request