The Role And Mechanism Of Heparanase In Invasion And Metastasis Of Thyroid Papillary Carcinoma

Objective: To investigate the expression of heparanase(HPSE)in papillary thyroid carcinoma(PTC)and its effects on the proliferation,cell cycle,apoptosis,migration,and invasion of PTC cells as well as the growth of PTC tumors.The molecular mechanism of PTC was analyzed to provide new ideas for the diagnosis and treatment of PTC.Methods:(1)HPSE m RNA expression in PTC tissues and cells was detected by RT-q PCR,HPSE protein expression in PTC tissues and cells was detected by Western Blot and immunohistochemistry,and the correlation between HPSE expression in PTC tissues and clinical features of PTC was analyzed.(2)HPSE overexpression plasmid and HPSE sh RNA interference plasmid were transfected into PTC cells B-CPAP and TPC-1.The transfection efficiency was verified by RT-q PCR and Western Blot.Cell proliferation was detected by Edu assay,cell apoptosis and cell cycle were detected by flow cytometry,and cell migration was detected by scratches assay.Cell invasion was detected by Transwell assay.The cells were subcutaneously injected into nude mice to observe tumor growth.The expression of tumor Ki-67 was detected by immunofluorescence.The expression of WNT/β-catenin signaling pathway-related protein was detected by Western Blot.(3)After HPSE was knocked down by B-CPAP and TPC-1,β-catenin was overexpressed in PTC cells.Protein expression of WNT/β-catenin signaling pathway was detected by Western Blot,cell proliferation was detected by Edu assay,apoptosis and cell cycle were detected by flow cytometry.Cell migration was detected by scratch assay,cell invasion was detected by Transwell assay.The cells were subcutaneously injected into nude mice to observe tumor growth.The expression of tumor Ki-67 was detected by immunofluorescence.Results:(1)The expression levels of HPSE m RNA and protein in PTC patients’ cancer tissues are higher than those in adjacent tissues,and HPSE expression level is positively correlated with PTC tumor size,invasion,and lymph node metastasis.Compared with thyroid follicular cell HT-ORI3,HPSE m RNA and protein expression levels in PTC cells B-CPAP and TPC-1 were increased.(2)Overexpression of HPSE increased the percentages of B-CPAP and TPC-1 Ed U positive cells in PTC cells,the percentages of G1 phase to S phase transition cells,the rate of scratch healing and the number of Transwell cells,decreased apoptosis rat,and increased expression of β-catenin and p-GSK-3β protein.Overexpression of HPSE also increases the size,weight,and volume of transplanted tumor,increases the expression of Ki-67.On the contrary,HPSE knockdown reduces the percentage of B-CPAP and TPC-1 Ed U positive cells,the percentage of G1 phase to S phase transition cells,the rate of scratch healing and the number of Transwell penetrating cells,increased apoptosis ratio,decreased expression ofβ-catenin and p-GSK-3β protein.The tumor size,tumor weight and tumor volume of nude mice were decreased,the expression of tumor Ki-67 was decreased.(3)HPSE knockdown can reduce the percentage of B-CPAP and TPC-1 Ed U positive cells in PTC cells,the percentage of G1 phase to S phase transition cells,the rate of scratch healing,the number of Transwell piercing cells,increased apoptosis rate,decreased tumor size,tumor weight and tumor volume,decreased tumor Ki-67 expression.HPSE knockdown and β-catenin overexpression increased the percentage of Ed U positive cells in PTC cells,the percentage of G1 phase to S phase transition cells,scratch healing rate and Transwell cell number.The expression of Ki-67 in nude mice were increased.Conclusion:(1)HPSE is highly expressed in PTC tissues and cells,and its expression level is positively correlated with tumor size,invasion,and lymph node metastasis of PTC patients.(2)HPSE promotes PTC cell proliferation,migration,invasion and tumor growth,and inhibits apoptosis.(3)HPSE promotes PTC cell proliferation,migration,invasion and tumor growth through Wnt/β-catenin signaling pathway,and inhibits apoptosis.