1.Differential Proteomic Analysis of Metastasis-associated Proteins in Mice Melanoma
Xinchao BAN ; Man LI ; Yanjun GU ; Dan LOU ; Xiuping WEI ; Xiulan ZHAO ; Baocun SUN
Chinese Journal of Clinical Oncology 2010;37(5):246-249
Objective: To investigate differentially expressed protein profiles in B16-F10 grafted melanoma and its metastasis in the lung in order to identify molecular markers of melanoma metastasis. Methods: Differentially expressed proteins in B16-F10 grafted melanoma and its metastatic lesion in the lung were isolated and identified by fluorescence two-dimensional differential gel electrophoresis(2D-DIGE)coupled with matrix assisted laser desorption ionisation time-of-flight mass spectrometry(MALDI-TOF-MS).Some of identified proteins were further confirmed by Real-time PCR analysis. Results: High resolutional images of differential gel electrophoresis were obtained and 9 of 30 differentially expressed proteins (IRatiol≥2,P<0.01)were identified by MALDI-TOF-MS.The expression of Myoglobin(MB),vimentin(VIM),phosphoglycerate kinase 1(PGK1),Triosephosphate isomerase(TPI or TIM),heavy-chain binding protein(BiP),α-enolase,β-actin,γ-actin,and laminin-binding protein were up-regulated in the experimental group compared with the control group.These proteins were involved in the cytoskeletal formation,glycolysis and so on.Real-time PCR analysis showed up-regulation of mRNA expression of PGK1 and TPI in the experimental group(P=0.001 and 0.003),which was in consistent with the resuits of proteomic analysis. Conclusion: A variety of abnormally expressed proteins contribute to the metastasis of mice melanoma.Glycolytic enzymes PGK1 and TPI may be involved in this process.
2.The Influence of SYT-SSX Fusion Gene, E-cadherin and β-Catenin on the Metastasis of Synovial Sarcoma
Xiuping WEI ; Yan SUN ; Xiulan ZHAO ; Wenjuan CAI ; Wenbin XIA ; Dan LOU ; Xinchao BAN ; Baocun SUN
Chinese Journal of Clinical Oncology 2010;37(4):205-208
Objective: To study the prognostic significance of the subtype of SYT-SSX fusion gene, E-cadherin, β-Catenin and clinicopathologicel parameters for the metastasis of synovial sarcomas. Methods: A total of 98 synovial sar-coma patients with complete clinical and follow-up data were reviewed. RT-PCR was used to detect the subtype of SYT-SSX fusion geneo The expression of E-cadherin and β-catenin was detected by immunohistochemistry. Univariate and multivariate analyses were performed to analyze the influence of the above factors and clinicopathological parameters on the metastasis free survival to explore the factors affecting the metastasis of synovial sarcoma. Results: Of all the pa-tients, 69.4% (68/98) had metastasis during follow-up. The median metastasis free survival was 48 months. The metastasis free 1-, 2-, 3-, 4-, and 5-year survival rate after surgery was 97.5%, 75.5%, 63.5%, 54.0%, and 48.5%, respectively; 31.6% (31/98) patients were found with SYT-SSX1 and 68.4% (67/98) patients with SYI-SSX2. The positive rate of E-cadherin ex-pression was 38.8% (38/98), the positive rate of β-catenin expression was 39.8% (39198) on cellular membrane and 53.1% (52/98) in cellular nucleus/cytoplasm. Univariate analysis showed that age (P=0.003), mitotic figure (P=0.002), histological grade (P=0.001), the subtype fusion gene of SYT-SSX (P=0.014), E-cadherin expression (P=0.015) and β-catenin expres-sion on cellular membrane (P=0.020) were significantly correlated with metastasis free survival of synovial sarcoma pa-tients. Sex (P=0.190), tumor location (P=0.105), tumor size (P=0.180), histological type (P=0.354), necrosis (P=0.451), β-catenin expression in cell nucleus/cytoplasm (P=0.911), radiotherapy (P=0.193), and chemotherapy (P=0.249) had no sig-nificant correlation with metastasis free survival of synovial sarcoma patients. Multivariate analysis revealed that the sub-type of SYT-SSX1 fusion gene (RR=2.505, P=0.003), negative expression of E-cadherin (RR=3.282, P=0.000), patient age (RR=2.157, P=0.004), and grade Ⅲ (RR=1.784, P=0.030) were independent risk factors for metastasis of synovial sarco-ma. Conclusion: The subtype of SYT-SSX, expression of E-cadherin, histological grade and the age of patients are impor-tant factors for evaluating the metastasis and prognosis of synovial sarcoma.
3.The Influence of Cell Proliferation and Microvessel Density on the Recurrence of Synovial Sarcoma
Wenbin XIA ; Yan SUN ; Xiulan ZHAO ; Xiuping WEI ; Wenjuan CAI ; Dan LOU ; Xinchao BAN ; Baocun SUN
Chinese Journal of Clinical Oncology 2010;37(4):201-204
Objective: To study the prognostic significance of cell proliferation and apoptosis, MVD and clinicopathologi-cal parameters for the recurrence of synovial sarcoma. Methods: We analyzed the clinical and follow-up data of 56 synovial sarcoma patients without metastasis. RT-PCR was used to detect the subtype of SYT-SSX fusion gene. The expression of Ki67 and MVD was detected by immunohistochemistry. Univariate analysis was employed to analyze the influence of the above factors and clinicopathological parameters on the recurrence free survival and to explore the influencing factors for the recurrence of synovial sarcoma. Results: Of all the patients, 73.2% (41/56) had recurrence during the follow-up. The median recurrence free survival was 19.5 months. The recurrence free 1-, 2-, 3-, 4-, and 5-year survival rates after surgery were 45.0%, 41.0%, 34.0%, 28.0%, and 28.0%, respectively. Ki-67 labeling index (LI) was 19.98%±11.64% and MVD was 51.83±21.92 per ×400. There was no significant difference in apoptotic index (AI) between the two groups (P=0.607). Χ~2 analysis showed that histological type (P=0.000) and MVD (P=0.045) were significantly correlated with the recurrence of sy-novial sarcoma. Univariate analysis showed that Ki67 LI (P=0.009), histological type (P=0.012) and radiotherapy (P= 0.014) were significantly correlated with the recurrence free survival of synovial sarcoma patients. Sex (P=0.015), tumor lo-cation (P=0.411), tumor size (P=0.801), necrosis (P=0.486), MVD (P=0.454), chemotherapy (P=0.272), and apoptotic grade (P=0.899) were not correlated with the recurrence free survival of synovial sarcoma patients. Multivariate analysis re-vealed that higher expression of Ki67 (RR=1.944, P=0.045), radiotherapy (RR=0.482, P=0.04), and histological type (RR= 0.207, P=0.031) were independent risk factors for the recurrence of synovial sarcoma. Conclusion: The expression of Ki67, radiotherapy and histological type are important factors for evaluating the recurrence and prognosis of synovial sarcoma.
4.Pilot Study on the Relationship of LPPCN with Neovascularization and Prognosis of Melanoma
Dan LOU ; Chunrong HAN ; Xiulan ZHAO ; Xiuping WEI ; Xinchao BAN ; Baocun SUN
Chinese Journal of Clinical Oncology 2010;37(2):80-84
Objective: To study the relationship of LPPCN with neovascularization and to analyze its clini-copathologic significance, in an effort to find a new target for anti-vascular therapies. Methods: Sixty-eight ma-lignant melanoma specimens were analyzed to observe the distribution of LPPCN and to examine the expres-sion of CD105 and TGFβ1 using immunohistochemistry. The distribution of vasculogenic mimicry (VM) was observed by immunohistochemical and histochemical double staining of CD31 and PAS. Results: (1) The tu-lines and networks. Of the 68 cases of melanoma, 55.89% (38/68) were recognized as having LPPCN. (2) In malignant melanoma specimens, the rate of vasculogenic mimicry density (VMD) and microvessel density (MVD) labeled by CD105 in LPPCN-positive group were higher than those in LPPCN-negative group, with sig-nificant differences (P<0.05). VMD and MVD were positively-correlated with the density of LPPCN. The posi-tive expression of TGFβ1 in LPPCN-positive group was higher than that in LPPCN-negative group and its ex-pression in the regions of LPPCN was obviously higher than that in circumambient tumor cells, with a signifi-cant difference (P<0.05). The expression of TGFβ1 was positively correlated with MVD labeled by CD105. (3) There was no relationship between LPPCN and gender, age, site, tumor embolus, lymph node metastasis or distant metastasis (P>0.05), but LPPCN was correlated with tumor size, mitosis figure count and Breslow depth (P<0.05). Kaplan-Meier survival analysis showed the survival rate of patients with LPPCN was lower than that of patients without LPPCN, with a statistical significance (P<0.05). The presence of LPPCN indicat-ed poor prognosis. Conclusion: LPPCN exists in malignant melanoma and is associated with VM and angio- genesis. Some tumor cells undergoing LPPCN have a spacial foundation for VM and angiogenesis. LPPCN can be an index for the evaluation of the prognosis of melanoma patients.
5.The classification and clinicopathological analysis of hepatocellular adenoma based on immunohisto-chemistry
Pengyan WANG ; Huilin SHAO ; Xinchao BAN ; Xiaoyan CHANG
Chinese Journal of Hepatobiliary Surgery 2019;25(7):509-512
Objective The subtypes of hepatocellular adenoma (HCA) were classified by immuno-histochemical study, and the clinicopathological characteristics of each subtype were analyzed. Methods From December 2003 to March 2018, 31 cases with HCA were retrieved from the archive files of the Depart-ment of Pathology, Peking Union Medical College Hospital, including 16 male patients and 15 female patients. The age ranged from 16 to 63 years. Hematoxylin and eosin ( HE) and immunohistochemical staining were performed with HCA samples. The subtypes were classified by immunohistochemical staining, and the clinicopathological characteristics of each subtype were analyzed. Results The HCA patients had no obvious and specific clinical symptoms, and most of them were diagnosed during the routine health checkup. All of the 31 patients were treated with surgery, and the complete resection was achieved in 26 cases. The adenomas were mainly in the right lobe of the liver (51. 6% , 16/31), and the solitary adenoma accounted for 54. 8% (17/31). 8 HCA were classified as H-HCA, 14 as I-HCA, 7 as β-HCA, and 2 as U-HCA by immunohistochemistry. H-HCA was characterized histologically by marked steatosis and lobulated contours, lacking L-FABP staining. I-HCA exhibited inflammatory infiltrate, telangiectasia, thickened arteries, more or less obvious ductular reaction, with the positive L-FABP/SAA/CRP staining. β-HCA all showed fibrous capsules, some of adenomas exhibited pseudoglandular structure and nodules in nodule. A strong homogeneous cytoplasmic overexpression of GS and nuclear β-catenin were observed in all β-HCA cases. The staining of SAA/CRP/GS was lacking in the U-HCA. Conclusions HCA is rare and difficult to diagnose in clinic. Final diagnosis relies on histological features, and immunohistochemical examinations need to be used for subtyping classification. Each of the four subtypes has characteristic pathological features.