| Backgroud:Pancreatic cancer occupy only1%to2%of whole body of the malignant tumor, but became10kinds of one of the most common malignancies, malignant tumor in the fourth to the cause of death.in spite of some consensus were be make in medical treatment of pancreatic cancer, including surgery surgical treatment, drug therapy and other auxiliary treatment, but the effect of treatment remained poor, and the mortality is so high that the5-year survival rate is about5%, be name by "the21st century stubborn fortress". There are still has many puzzle in the occurrence and development mechanism of Pancreatic cancer, if we can find the the target of cancer happens, and interfere with the target to suppress the occurrence or development speed of pancreatic cancer, to provide the theory basis of pancreatic cancer treatment.Mir-146a is a22nt non-coding RNA, there are widely exists in plants and animals of somatic cells, it choose effect to mircroRNA target genes, restrain target genes transcription or degradation target genes. Many studies show that, Mir-146a expression Obvious relation with breast cancer, ovarian cancer, pancreatic cancer and was associated with oral cancer. In Targetscan have found that smad4and timp3are both target genes of Mir-146, and my mircroRNA research team at chip analysis discovered that, Mir-146a in pancreatic cancer cells express higher than in normal pancreatic cells, this signal cause our attention, smad4and timp3are both take an important role in the development of tumors,Smad4usually low expression in pancreatic cancer, colon cancer, some scholars repair its expression, observed that Smad4main control tumor cell proliferation and invaded and metastasis. Timp3in liver cancer, small cell lung cancer, breast cancer plays a role of inhibiting tumor cell invaded and apoptosis. In the basis of our research, preliminary validation smad4and timp3whether or not a target gene of Mir-146a, and combined with clinical pathologic features, observed the relation of smad4or timp3expression with clinical pathologic features, whether have some clinical significance.Objective:This experiment verified through the pancreatic cancer Timp3and Smad4protein expression, and indirect test Timp3and Smad4gene is a target genes of Mir-146a, and through the molecular expression of Timp3and Smad4, analysis the correlation of pathologic characteristics and prognosis of the patient, in order to close the research work of pancreatic cancer and clinical application.Method:By our research team selection14cases specimens of ductal carcinoma of the pancreas and6cases of normal pancreas tissue, send to Shanghai biological chips for MircroRNA genetic analysis, discover that mir-146a expression in pancreatic cancer groups5.6times higher than normal pancreas tissue. Use Targetscan gene database predicted that Smad4and TIMP3may be both target genes of mir-146a, mir-146a through3’UTR extremity don’t fully complementary combined with Smad4or TIMP3, restrain expression of Smad4or TIMP3.We use immunohistochemical method, According to0-3level to comprehensive description of the dyeing strength and dyeing rate.0:no dyeing or positive cells<10%,1:>10%of tumor cells positive or10-40%of the tumor cells dyeing weak positive,2:>40%tumor cells dyeing, weak positive or10-40%tumor cells strong dyeing,3:>40%of tumor cells strong dyeing.half quantitative detection Smad4and Timp3molecular’s expression of pancreatic cancer and normal pancreas, indirect test and verify Smad4and Timp3weather or no a target gene of mir-146a, and by increasing8cases of pancreatic cancer immunohistochemical (22cases of pancreatic specimens), through the regular long-term follow-up datas, analysis the relationship between Smad4and Timp3protein expression and tumor of pathologic characteristics and the survival in Pancreatic cancer, finding each factor of the influence for the prognosis of pancreatic cancer surgery. All statistical analyses were performed using the SPSS13.0statistical software package. The Chi-square test, Fisher’s exact test and Mann-Whitney U test were used to assess the relation between the proteins expression and clinicopathologic parameters. Overall survival curves between subgroups divided according to the proteins expression levels were drawn by using the Kaplan-Meier method, and significant differences among subgroups were compared by using the log-rank test. Multivariate analyses were performed using Cox proportional hazards model to identify independent prognostic factors. a=0.05(two-sided) as the difference level, and P<0.05was considered indicative of statistical significance.Result:1. According to14cases of pancreatic cancer and6cases of normal pancreatic tissue immunohistochemical, Smad4and Timp3expression in normal pancreatic tissue obviously higher than in pancreatic cancer (P=0.002, P=0.008), mir-146a and Smad4protein’s expression significant correlation (r=0.741, P<0.001), but the negatively related. mir-146a and Timp3protein’s expression significant correlation (r=0.485, P=0.03), and negatively related.2. Smad4protein’s expression have no significant relationship with pancreatic cancer patients gender (P=0.662), age (P=0.091), smoking or/and drinking (P=0.386), PET’s SUVmax value (P=0.07), tumor size (P=0.183), but have significant relationship with tumor’s differentiation degree (U= 30.0, P=0.015), T stage (P=0.024), N stage (P=0.006) and the clinical stage (U=22.5, P=0.012).3. Timp3protein’s expression have no significant relationship with pancreatic cancer patients’ gender (P=0.666), age (P=0.67), smoking or/and drinking (P=0.378), PET’s SUVmax value (P=0.179), tumor size (P=0.67), cancer pathological differentiation degree (U=45.0,P=0.177), T stage (P=1.000), N stage (P=0.391) and the clinical stage (U=49.0, P=0.403).4. The survival of the single factor analysis showed that the survival have no significant relationship with gender (χ2=1.744, P=0.187), age (χ2=2.563, P=0.109), smoking or/and drinking (χ2=0.077, P=0.781), Timp3protein’s expression (χ2=1.015, P=0.314). but have significant relationship with PET SUVmax value (χ2=11.836, P=0.001), tumor size (χ2=9.736, P=0.002) pathology differentiation degree (χ2=5.21, P=0.022), Smad4protein’s expression (χ2=5.04, P=0.025), T stage (χ2=5.854, P=0.016), N stage (χ2=4.43, P=0.035) and the clinical stage (χ2=5.854, P=0.016) in pancreatic cancer postoperation patients.5. Cox regression analysis found that pancreatic cancer pathological differentiation degree (R=1.063, P=0.016) and the tumor size (R=0.155, P=0.019) are the independent factors affecting the prognosis.Conclusion:1. Mir-146a have significant correlation with Smad4expression, Smad4may be a target gene of mir-146a, but have no significant correlation with Timp3expression,Timp3may not a target gene of mir-146a.2. Smad4protein expression have significant relationship with pancreatic cancer pathology differentiation degree, T stage, N stage and the clinical stage, detection Smad4protein expression in pancreatic cancer patients, which can provide a judgment of important clinical indicators of pancreatic cancer like pathological differentiation degree, T stage, N stage, clinical stage, prognosis, etc.3. The survival have obvious relationship with PET’s SUVmax value, tumor size, pathological differentiation degree, Smad4protein expression, T stage, N stage and the clinical stage in pancreatic cancer postoperation patients. independent predictors of survival were tumor size and tumor differentiation. |
PDF Full Text Request