The Clinical And Experimental Research Of The Function And Significance About Th17、 CD40、 PCNA In Sacral Chordomas

Purpose: To discuss the effect of Th1、Th2、Treg、Th17 in the occurrence and development of sacral chordoma, and the relationship with biological behaviour of sacral chordoma, and provide some theoretical basis for the diagnosis, prognostic evaluation and immunological targeted therapy of sacral chordoma.Methods: To collect the fresh tumor tissue of 27 patients with sacral tumors who received surgery in our hospital from January 2002 to December 2008. Among them, there were 11 patients with sacral chordoma, 9 with giant cell tumor of the sacrum, 7 with sacral nerve sheath tumor. Using Real-time PCR to detect the expression of FoxP3 mRNA, ROR-γt mRNA、T-bet mRNA and GATA-3 mRNA, using mean±standard error to represent the expression of all genes, and using Wilcoxon rank test to compare the expression of genes. It has statistical significance if P <0.05.Results:①The relative expression of FoxP3 in sacral chordoma, giant cell tumor of the sacrum and nerve sheath tumor were 8.5±7.6%、1.6±0.9%、0.7±0.5%. The expression of FoxP3 in sacral chordoma was higher than that in giant cell tumor of the sacrum and nerve sheath tumor. The difference has statistical significance (P<0.05).②The The relative expression of T-bet in sacral chordoma, giant cell tumor of the sacrum and nerve sheath tumor were 0.7±0.3%、1.7±1.2%、1.6±1.3% . The expression of T-bet in sacral chordoma was lower than that in giant cell tumor of the sacrum and nerve sheath tumor. The difference has statistical significance (P<0.05).③The relative expression of GATA-3 in sacral chordoma, giant cell tumor of the sacrum and nerve sheath tumor were 2.9±2.3%、0.6±0.5%、0.3±0.1%. The expression of GATA-3 in sacral chordoma was higher than that in giant cell tumor of the sacrum and nerve sheath tumor. The difference has statistical significance (P<0.05).④The relative expression ROR-γt of in sacral chordoma, giant cell tumor of the sacrum and nerve sheath tumor were 4.1±2.9%、1.6±0.8%、0.2±0.1%.The expression of ROR-γt in sacral chordoma was obviously higher than that in giant cell tumor of the sacrum and nerve sheath tumor .The difference has statistical significance (P<0.05).Conclusion: The high-expression of FoxP3 mRNA、GATA3 mRNA and low- expression of T-bet mRNA in the primary sacral chordoma patients indicated that the local tissue of the tumors appeared migration phenomenon of Th2, and it may be one of the reasons for the immunologic escape. The increased expression of ROR-γt mRNA indicated that there was advantage phenomenon of Th17 in the primary sacral chordoma tissue, and it may cause the recurrence of chordoma by upexpressing the angiogenic factors of the tumors.Part two The study of Th17 and its relationship with prognosis in primary sacral chordomaPurpose: To discuss the correlation between Th17 and the recurrence after operation, survival time without tumor, overall survival time. And to provide some reference for prevent or delay the recurrence of sacral chordoma after operation, prolong the survival time without tumor after operation in the future.Methods: To follow up the 28 patients with sacral chordoma who received the first operation in our orthopaedic department from January 1998 to December 2008. Using the immunohistochemisty with the pathology wax. Besides, using the immunity study for the 16 fresh tissue of giant cell tumor of the sacrum and nerve sheath tumor which were stored in the liquid nitrogen. Using Kaplan-Meier to get the single factors analysis of variance and draw the curve of CDFS, and using the Log-Rank to check the difference of recurrence. It has statistical significance if P <0.05.Results:①The positive expression of Th17 in sacral chordoma was 60.7%,and weakly positive expressed in group of giant cell tumor of the sacrum, no expression in group of nerve sheath tumor.②There were no statistical significance for the expression of Th17 in sacral chordoma between different age and gender (P>0.05).③The positive expression percentage of Th17 in the group of tumor diameter≥10cm was 83.33%,for the group of tumor diameter<10cm was 40.75%,there was statistical significance (P>0.05).④The positive expression percentage of Th17 in the group of positive incisal margin was 69.23%, which was higher than that of negative incisal margin(53.33%), but no statistical significance(P>0.05).⑤The positive expression percentage of Th17 in the group of recurrence was 78.57% and 42.86% in the group of no-recurrence. There was statistical significance (P<0.05). The survival rate of 5-year no recurrence in the group of positive Th17 was (32.4±12.8)%, overall survival rate of 5-year was (86.9±8.7) %. The survival rate of 5-year no recurrence in the group of negative Th17 was (81.8±11.6)%, overall survival rate of 5-year was 100 %. Conclusions: Th17 played an important role in the local recurrence of sacral tumors.The high expression of Th17 indicated the high recurrence and short CDFS after operation. The high expression of TH17 was an important sign for the progress and recurrence,and could be one of the indexs to determine the recurrence of the sacral chordoma. Purpose: To study the expression level of CD40、PCNA in sacral chordoma and their relationship with recurrence, and to provide a new target for the immunization therapy of sacral chordoma.Methods: To collect tumor tissue of 28 cases with sacral chordoma who had full information of clinical and pathology for the wax. To detect the expression of CD40 and PCNA by using immunohistochemisty, another 9 cases with normal notochordal tissue were set as control group and count the MVD marked by CD31. All the data was showed by mean±standard error and using T-test to compare two groups. It has statistical significance if P <0.05.Results:①The strong positive expression of CD40、PCNA in the sacral chordoma were 66.1% and 64.3%, obviously higher than that in normal notochordal tissue. There was statistical significance (P<0.05). The MVD marked by CD31 in sacral chordoma was (24.26±18.68)/mm2 and (4.63±2.19)/mm2 in normal notochordal tissue. There was statistical significance (P<0.05).②Among the 28 sacral chordoma, 14 had recurrence and 14 had no recurrence, the positive expression of CD40 were 14.29% and 85.71%.For the PCNA, the positive expression were 92.86% and 50.55%. MVD marked by CD31 were (30.66±23.75)/mm2 and (17.88±8.52)/mm2. All had statistical significance (P<0.05).③There were no statistical correlation between the expression of CD40、PCNA and the value of MVD with the age, gender and the size of tumor.④The MVD in the group of positive expression of PCNA was (28.92±20.27) /mm2, significantly higher than the negative group (12.64±3.70) /mm2. And there was statistical significance(P<0.05). The MVD in the group of negative expression of CD40 was (31.68±22.81 /mm2, significantly higher than the positive group (16.86±9.28) /mm2. And there was statistical significance (P<0.05).Conclusions: CD40 and PCNA had vital role in the tumor invasion, recurrence and metastasis. Strong expression of PCNA and weak expression of CD40 was important sign for the progress and recurrence of sacral chordoma. The expression level of CD40 and PCNA in the sacral chordoma was one of the index to determine the progress and recurrence of the tumor.