The Mechanism Of Chitinase 3 Like Protein 1 In Gallbladder Cancer

Part Ⅰ The expression of Chitinase 3 like protein 1 in gallbladder cancerObjective:To detect the expression of Chitinase 3 like protein 1 in gallbladder cancer samples. To analyze the relationship between Chitinase 3 like protein 1 expression and the pathological characteristics of gallbladder cancer.Methods:We test the expression of CHI3L1 in 65gallbladder cancer tissues and 20 cases of chronic cholecystitis gallbladder mucosal tissue. All calculations were performed with SPSS 17.0 for Windows. Pearson χ2 test or Fisher exact test was used to compare qualitative variables. Kaplan-Meier analysis was used to determine survival.Result:CHI3L1 was highly expressed in gallbladder cancer tissues in the cytoplasm. The positive rate was 87.7%, the expression of CHI3L1 protein in chronic cholecystitis was weak, and the positive rate was 11.1%. Clinic pathological analysis showed that positive CHI3L1 expression was significantly associated with growth pattern, the amount of transferred lymph nodes, distant metastasis and TNM stage (P< 0.05, respectively). Survival analysis showed that the positive expression of CHI3L1 protein in survival time was shorter than the negative CHI3L1 expression in patients with advanced carcinoma of gallbladder, the difference was statistically significant (P< 0.05).Conclusion:CHI3L1 was highly expressed in gallbladder cancer tissues, the expression of CHI3L1 was closely associated with the stage of cancer and lymph node metastases, and it may play an important role in the invasion and lymphatic metastases of the tumor. CHI3L1 was closely associated with poor prognosis in patients with gallbladder cancer.Part Ⅱ Influence of gallbladder cancer cells after restraining the expression of CHI3L1 by RNA interference in vitroSection Ⅰ Construction of RNA interference lentvirus vector targeting CHI3L1Objective:To study whether siRNA-CHI3L1 could inhibit the CHI3L1 gene expression in gallbladder cancer cells.Methods:Lentiviral-mediated CHI3L1 RNA interference was constructed. We examined the knockdown of CHI3L1 by siRNA CHI3L1 mRNA express by RT-PCR and the CHI3L1 protein express by Western-Blot in gallbladder cancer cells.Results:The CHI3L1mRNA express and its protein express decreased significantly in GBC-SD gallbladder cancer cells by RNA interference (P<0.05).Conclusion:RNA interference expressing vector targeting CHI3L1 was successfully constructed.Section Ⅱ Influence gallbladder cancer GBC-SD cells after CHI3L1 silencing in vitroObjective:To study the effect of RNA interference of CHI3L1 in the proliferative, invasion and migratory activities of human gallbladder cancer cells.Methods:Human gallbladder cancer cells GBC-SD were divided into 3 groups:GBC-SD group, siRNA-CHI3L1 group and siRNA-negative control group. The proliferative, migratory and invasion activities of every group were detected using cell active assay (CCK-8) and transwell assay; the MMP-9 was detected by Elisa test.Result:The CHI3L1 accelerate the cell proliferation and enhance the migration, invasion ability; it also enhances the secretion of MMP-9. They were decrease in migration, invasion activities in GBC-SD (siRNA-CHI3L1), also the secretion of MMP9 decreased in GBC-SD (siRNA-CHI3L1).Conclusion:The CHI3L1 protein enhanced the GBC-SD cells migratory and invasion behavior by secretion of MMP-9. And it also could enhance the cell proliferation of GBC-SD.Part Ⅲ The effect of CHI3L1 activation on the cellular signal pathwayObject:To study the cellular signal pathway activated by CHI3L1, and investigates the mechanism of CHI3L1 enhancing migration and invasion of GBC-SD cells.Methods:We collected the cells lyses of the three groups’cells, and tested the function expression of ERK1/ERK 2 and JNK by Western-Blot. And the invasion function of GBC-SD was test with U0126 (the inhibitor of MEK-ERK).Result:The functional ERK1/ERK2 and JNK reduced in the siRNA-CHI3L1 group cells. And the U0126 could inhibit migration and invasion function of GBC-SD cell.CHI3L1 induces IL-8 and MCP-1 secretion from gallbladder cancer cells may via ERK and JNK.