Expression Of Syk In Human Glandular Epithelium Tumor Cell By Preoperative Chemotherapy And Clinicopathological Features

Part 1 Expression of Syk in different origins of human glandular epithelium tumor cell一,Expression of Syk in different origins of human glandular epithelium tumor cellObjective:To study the expression of Syk in the human glandular epithelium tumor cell line.Methods: To detected the expression of Syk with immunohistochemical staining in human glandular epithelium tumor cell lines.Results: Human breast cancer cell line: Syk expression was showed in MCF-7 cells, but not showed in MDA-MB-231 cells. Syk was positive expressed in human pancreatic cancer cell line PANC-1,SW1990 and Capanc-2 and human adenocarcinoma of lung cancer cell line GLC-82 and human hepatocellular carcinoma cell line SMMC-7721.Conclusion: Syk was positive expressed in the human glandular epithelium tumor cell line. Syk expression is the effective index for the prognosis of human glandular epithelium tumor.二,Expression of Syk in human breast cancer / pancreatic cancer tissue and clinical featureObjective:To study the expression of Syk in the human breast cancer/ pancreatic cancer tissue and clinical featureMethods: To detected the expression of Syk with immunohistochemical staining and real-time polymerase chain reaction in 30 pairs of human breast cancer and 22 cases of pancreatic cancer tissue and normal issues and clinicopathological features was analysis. Results: Reduced expression level of sykmRNA and protein has been noted in breast carcinoma tissue ( 63.3%,60%) compared to normal breast tissue (93.3%,90%) (P<0.01). The lower expression levels in breast carcinoma tissue was related with clinicopathological features including lymph-node metastasis and TNM stage (P<0.05), but not with the histological grade nor the estrogen and progesterone receptor status (P>0.05). In breast carcinoma tissues, the level of syk mRNA is (6.17±0.0.65)×10⁴ is lower than the level in the normal tissues (80.37±0.125)×10⁴ (P<0.01). Compared to normal tissue (90.9%,86.3%),pancreatic cancer tissue ( 63.6%,54.5%) as reduced the level of expression sykmRNA and protein (P<0.01). The mean level of sykmRNA of normal pancreatic tissue and cancer is(52.9±0.026)×10⁴,(4.45±0.225)×10⁴ respectively(P<0.001);The lower levels in pancreatic cancer tissue was related with clinicopathological features including lymph-node metastasis and TNM stage (P<0.05), but not tumor size and histological grade (P>0.05). The positive expression rate of p53 in pancreatic cancer tissue is 13/22; The positive expression rate of p53 in pancreatic cancer tissue with celiac lymph nodes is 8/16; It is 5/6 in non- celiac lymph nodes. The sykmRNA positive expression of 10 case in 13 of p53 positive expression. The relation of syk and p53 is direct correlation(r=0.8531);The average live time of Syk⁺⁺⁺,Syk⁺⁺,Syk⁺,Syk^- in pancreatic cancer is 12,10.3,5.6,1.8months (P<0.01). The more syk positive expression, The more the mean live time.Conclusion: The expression of sykmRNA and protein was reduced in human invasive breast carcinoma/ pancreatic cancer. Down-regulated expression of Syk might be assiociated with tumor invasion and metastasis. Part 2 Expression of Syk in human glandular epithelium tumor tissue by preoperative chemotherapy and clinicopathological features一,Expression of Syk in human breast cancer tissue with lobaplatin by preoperative chemotherapy and clinlcopathological features(一)Pharmacokinetic Study with Lobaplatin in patients with breast cancerObjective: To study the pharmacokinetics of lobaplatin in Chinese cancer patients.Methods: Eight patients entered the study. The dose of lobaplatin is 50mg/m2 with i.v. continuous jnfusion for 2 hours. The blood and cancer tissue of patients were sampled for the determination of intact lobaplatin. Lobaplatin concentration were measured by high-performane liquid chromatograpy with ultraviolet detection.The pharmacokinetic parameters were calculated by 3p97 PKS software.Results: The mean concentration of ten dots after end of lobaplatin infusion at t=0 and 0.2,0.5,1,2,4,6,8,12,24 hour is 26.26±3.10,16.83±2.00,11.24±1.63,6.43±1.45,4.45±1.57,2.19±0.26,1.53±0.25,0.79±0.18,0.47±0.11,0.02±0.01mg·L^-1 respectively;C_max:25.56±3.04mg·L^-1;T_1/2α0.25±0.04 hour;T_1/2β2.70±0.25hour;K211.00±0.28/h;K100.74±0.11/h;K₁₂1.31±0.27/h;AUC35.20±5.47μg·h·L ^-1;CL8.73±1.51L·h^-1。Conclusion: Pharmacokinetics of lobplatin fit biphasic kinetic models. The pharmacokinetic parameters of lobaplatin and its metabolite are similar comparison with that reported of lobaplatin in patient's plasma. There is a higher concentrations of lobaplatin distributed in the tumor tissue compared with that of in plasma.(二)Expression of Syk in human breast cancer tissue by Lobaplatin preoperative chemotherapy and clinicopathological featuresObjective: To detect blood target and immune function in breast cancer with lobaplatin by preoperative chemotherapy. Lobaplatin concentrations of blood plasma and cancer tissue were measured with high-performane liquid chromatograpy (HPLC). To observe the cancer cell morphology by H-E dyeing methods.To study the expression of Syk in human breast cancer tissue by preoperative chemotherapy and clinicopathological featuresMethods: Syk expression was detected with immunohistochemical staining and real-time polymerase chain reaction to evaluate the expression of syk and p53 in two groups .control group is 30 pairs of human breast cancer tissue and its surrounding tissues and two types breast cancer cell line MCF-7,MDA-MB-231 ;The experimental group with lobaplatin after 24 hour and clinicopathological features was analysis.Results: The mean of lobaplatin concentrations in cancer tissue after infusion 24 hours is 0.23±0.15mg·L^-1 ,more higher than that of in blood plasma( 0.02±0.01mg·L^-1()P<0.0001), lower than the effective concentration. There is no significant difference in blood routine,Biochemistry,clotting time and immune function; The expression of sykmRNA and protein in experimental group is significantly raise up in MCF-7,MDA-MB-231 cell line. The level of sykmRNA is from 1.41×10⁴ to 3.13×10⁴,0 to 1.11×10⁴ respectively;The positive expression of sykmRNA in control and experimental group is 63.3%(19/30),86.7%(26/30).The level is(6.17±0.065)×10⁴,(26.39±0.017)×10⁴(P<0.001).Compare to two groups in histopathology: There are a lot of eosinophlis were observed in the tumor tissues or around it of expression group. At the same time , the atrophy of the tumor cells can also be seen. The necrosis of the tumor cells were obviously. The red cytoplasm,pyknosis and karyolysis were taken as evidence of cell necrosis. Only a few of survival tumor cells were vacuole degeneration.Conclusion: The rate and level expression of sykmRNA and protein with lobaplatin as preoperative chemotherapy is significantly raise up.二,Expression of Syk in human pancreatic cancer tissue by Gemitabin preoperative chemotherapy and clinicopathological featuresObjective: To study the expression of Syk in human pancreatic cancer tissue by Gemitabin preoperative chemotherapy and clinicopathological features.Methods: To evaluate the expression of syk and p53 in human pancreatic cancer tissue by H-E methods and immunohistochemical staining by preoperative chemotherapy with Gemitabin and clinicopathological features was analysis.Results: The rate of positive expression of Syk is 92.3%,76.3% in the normal and cancer tissue of pancreatic patients in experimental groups respectively. That of in control is 89.2%,54.5%;It is significantly difference in the cancer tissue of two groups(P<0.01).The expression of p53 protein is 79.3%,59.9% in the experimental and control group respectively. Compare to the two groups in histopathology: The expression group is a lot of eosinophlis were observed in the tumor tissues or around it. At the same time, the atrophy of the tumor cells can also be seen. The necrosis of the tumor cells were obviously. The red cytoplasm,pyknosis and karyolysis were taken as evidence of cell necrosis. Only a few of survival tumor cells were vacuole degeneration. Compare to control, there is more positive Syk in experimental group , which is more deep and eribble in dyeing of Syk. The dyeing of cancer tissue of Syk is near to normal tissue.Conclusion: The rate of Syk expression is raised up by preoperative chemotherapy with Gemitabin in pancreatic cancer patient, improve the prognosis.Part 3 The mechanism study of Syk expression in human glandular epithelium malignant tumor tissue by preoperative chemotherapy一,The mechanism study of Syk methylation in breast cancerObjective: To study the mechanism of Syk methylation in breast cancer.Methods: The methylation in CpG of Syk promoter region is detected by using MSP methods and clinicopathological features was analysis. Results: There is no methylation in breast normal tissue, but 17 cases takes place methylation in 30 cases breast cancer tissue. The rate of methylation is 0,56.7% which is signficancely difference in breast normal and cancer tissue respectively(P<0.001).The rate of Syk methylation in stage of III is 41.6% which is more high than that of in stage II 22.2%(P<0.001). The methylation of Syk promoter is positive correlation with clinical stage of TNM, which is no correlation with tumor differentiation,size and tumor location.Conclusion: The methylation of CpG is one of reason of the Syk inactivation.二,The mechanism study of SykmRNA increasing with lobapltin by preoperation chemotherapy in breast cancerObjective: To study the mechanism of Syk methylation in breast cancer and the increasing of SykmRNA with lobaplatin by preoperation chemotherapy in breast cancer.Methods: To detect the methylation in CpG of Syk promoter region with MSP methods and to analysis clinicopathological features. To detect the expression of Syk,IL-2Rβ,CD4 and c-myc in two groups of breast cancer and study the relationship.Results: There is no methylation in breast normal tissue, but 17 cases take place methylation in breast cancer tissue. The rate of methylation (0,56.7%) is signficancely difference in normal and cancer tissue (P<0.001).The rate of Syk methylation in stage of III(41.6%) is more high than that of in stage II 22.2%(P<0.001). The rate of methylation of Syk promoter is positive correlation with clinical stage of TNM, non correlation with tumor differentiation,size and tumor location. The expression of Syk,IL-2Rβ,CD4 and c-myc is raised up in experimental groups, the rate of Syk and c-myc (86.7%,70%) is more high than that of in control groups. It is significationly difference in Syk and c-myc (P<0.05 );not in IL-2Rβand CD4 (P>0.05 ).Conclusion: The methylation of CpG is one of reason of the Syk inactivation. The increasing of Syk and c-myc expression can inhibited the tumor growth.