Effects Of SiRNA Knock-down Of Annexin A1 On Biological Characteristics By Carcinoma Of Papillary Thyroid In TPC-1 Cells

Objective:(1)To explore the correlations between Annexin A1 protein expres-sion and clinicopathological characteristics in carcinoma of papillary thyroid.(2)Detect the transfection efficiency of special interference RNA(siRNA) of Annexin A1 in TPC-1 carcinoma of papillary thyroid cells.(3)After special siRNAof Annexin Al was successfully transfected into TPC-1 carcinoma of papillary thyroid cells. Detect the Annexin Al expression of all groupas form mRNA level. To validate the special siRNA successfully knock-down Annexin A1 in TPC-1 carcinoma of papillary thyroid cells(4)Analyze the apoptosis efficiency, proliferation, invasion and cell migration of TPC-1 carcinoma of papillary thyroid cell after Annexin Al successfully been knock-down.To research the relationship between the of Annexin A1 and TPC-1 carcinoma of papillary thyroid cells.Methods:(1) Using immunohistochemistry to investigate investigate the different expressions of Annexin A1 in papilary thyriod tissue and para-carinoma tissue.(2) To use siDirect software(Version2.0) to design 3 fragments of the specific small interference ANXA1 gene (siRNA), respectively ANXA1-siRNA1, ANXAl-siRNA2, ANXAl-siRNA3 and negative-siRNA. To transfect siRNA into papillary thyroid carcinoma TPC-1 cells by Lipfectamine2000, detect the change of mRNA expression before and after the ANXA1 gene silencing in TPC-1 by RT-PCR after calculate the transfection efficiency.(3) When ANXA1 was knockdown, to detect the apoptosis of TPC-1 cells by RNA interference flow cytometry after 72h; CCK-8 Kit Method to assay the proliferation of papillary thyroid carcinoma TPC-1 cells after RNA interference of 24h,48 h,72h and 96h cell; Transwell cell migration, invasion of TPC-1 cells, RNA interference detection after the migration and invasion ability.Results:(1)Among 69 sample tissuesof papillary thyriod carcinoma,the positive rate of Annexin Al was higher than that of 69 para-carcinoma tissues (tumor tissuee vs para-carcinoma tissures(88.41%:8.69%),there was significant different (p<0.0001). Furthermore,the expression of Annexin A1 is related with the lymph node metastasis and tumor size.(2)After transfection, fluorescence microscope was used to observe and record the sirRNA transfection, and to calculate the transfection efficiency, the direct calculation method (repeated three times), the transfection efficiency was about 90%.(3) Papillary carcinoma of thyroid TPC-1 cells transfected with 72h, experimental group ANXA1-siRNA1, ANXA1-siRNA2, ANXA1-siRNA3, and blank control group, negative control group, liposome group,3 ANXA1 mRNA gene specific siRNA group decreased levels of expression, the ANX-siRNA3 decreased significantly.(4) When ANXA1 knockdown, apoptosis in TPC-1 papillary thyroid cancer cells compared with control group, negative control group increased; cell proliferation compared with blank control group, negative control group weakened; invasion and metastasis compared with the blank group and negative control group.Conclusion:This study demonstrated that in human papillary thyroid carcinoma, the positive rate of ANXA1 protein was significantly higher than that in paracancerous tissues. In papillary thyroid carcinoma TPC-1 cells, while ANXA1 knockdown, down regulation of ANXA1 expression. At the same time, the apoptosis induced by thyroid cancer TPC-1 cells inhibited cell proliferation, reduce cell invasion and metastasis. Thus the effect of TPC-1 cells of papillary thyroid carcinoma, apoptosis, invasion and metastasis ability of appreciation by the expression of ANXA1. Using ANXA1 as a biological target, and provides a new idea for the diagnosis of papillary thyroid cancer, treatment, and prognosis.