Study On The Mechanism Of JAZF1 On The Proliferation,Apoptosis,Invasion And Migration TPC-1 Thyroid Cancer Cell Line

Objective: To investigate the expression of JAZF1 in normal and papillary thyroid carcinoma cells,the effect and mechanism of JAZF1 on the proliferation,apoptosis,invasion and migration of papillary thyroid carcinoma TPC-1 cells.Methods: 1.Use real-time fluorescence quantitative polymerase chain reaction(q RTPCR)to detect m RNA,and protein expression levels of JAZF1 in normal thyroid cells and papillary cancer cells are tested by Western blotting(WB).Set up Adv-JAZF1-GFP recombinant adenovirus and infect the thyroid papillary carcinoma TPC-1 cell as the test group.Compare it with the blank control group(NC)and no-load group(Adv-GFP);2.Use CCK-8 method to detect the proliferation of TPC-1 cells in each group.3.Effects of JAZF1 on the invasion and migration of TPC-1 cells are tested by scratch experiment and Transwell.4.Measure the expression of related protein indicators in each TPC-1 cells group;Results:1.JAZF1 has the same expression trend in normal thyroid epithelial cells Nthyori3-1 and thyroid papillary cancer TPC-1 and BCPAP cells.The relative expression of BCPAP protein is 67% lower compared to Nthy-ori3-1,and the relative expression of TPC-1 protein it was down-regulated by 43%(P <0.05);the relative expression of BCPAP m RNA was down-regulated by 68%.The relative expression of TPC-1 m RNA was downregulated by 45,compared with Nthy-ori-3 %(P<0.05).2.The TPC-1 cell line with JAZF1 gene over-expression was successfully constructed.WB test results showed that the relative expression of JAZF1 protein in the Adv-JAZF1-GFP group of TPC-1 cells was 2.15 times higher than that in the NC group and 2.05 times higher than that in the Adv-GFP group Times(P<0.05);3.CCK-8 experimental results show that compared with the other two groups,the over-expressed JAZF1 can inhibit the proliferation of TPC-1 cells compared with the other two groups(P<0.05);4.Cell scratch test results show that the scratch distance of the Adv-JAZF1-GFP group measured at 12 h and 24 h after the scratch is significantly larger than that of the NC and Adv-GFP groups(P<0.05);5.Transwell results showed that the invasion and migration ability of the Adv-JAZF1-GFP group was significantly weaker than that of the NC group and the Adv-GFP group(P<0.05);6.WB detection and expression of cell proliferation-related proteins: the relative expression of tm TOR in the Adv-JAZF1-GFP group was reduced by 41% and 40% respectively compared with the NC group the Adv-GFP group(P <0.05);The relative expression in the AdvJAZF1-GFP group was reduced by 51% compared with the NC group and 48% compared with the Adv-GFP group(P <0.05);the relative expression of Raptor protein was not significantly different among the three groups(P>0.05);7.WB detection and expression of apoptosis-related proteins: the relative expression of Bcl-2 in the Adv-JAZF1-GFP group was reduced by 37% compared with the NC group and 30% compared with the AdvGFP group(P <0.05);The expression of Bax in the Adv-JAZF1-GFP group increased 1.4-fold compared to the NC group and 1.86-fold(P <0.05)compared to the Adv-GFP group;the expression of Caspase-9 in the Adv-JAZF1-GFP group increased 1.67 times compared with the NC group,and 1.58 times compared with the Adv-GFP group(P <0.05).The relative expression of Cyto-C in the Adv-JAZF1-GFP group increased 1.61 times compared with NC group,and it increased 1.47 times compared with Adv-GFP group(P <0.05);8.WB detected expression of MMP2 and MMP9 which are related to invasion and migration: The relative expression of MMP2 protein in the Adv-JAZF1-GFP group was decreased by 51% compared with the NC group and the Adv-GFP group(P <0.05);The relative expression of MMP9 protein in Adv-JAZF1-GFP group was reduced by 38% compared with NC group and Adv-GFP group(P<0.05)Conclusion: The expression of JAZF1 in papillary thyroid carcinoma cells is lower than that of normal thyroid epithelial cells.Overexpression of JAZF1 can inhibit the proliferation of TPC-1 cells,promote apoptosis,and inhibit cell invasion and migration.The mechanism may be related to the down-regulation of t-m TOR,Rictor,and Bcl-2.It is also related to the up-regulation of Bax,Caspase-9,and Cyto-C,while the weakening of invasion and migration ability may be related to the down-regulation of MMP2 and MMP9.