The Mechanism Of ANXA1 Expression And TPF Induction Chemotherapy In Oral Squamous Cell Carcinoma

Background and PurposePrevious experiments have shown that the ANXA1 plays an important role on progression of OSCC,and a retrospective study has found that patients with locally advanced OSCC could benefit from TPF induction chemotherapy in moderate/poorly pathologic differentiation grade and low expression of ANXA1.Howerver,the mechanism of ANXA1 in OSCC is still not clear.This study aims to clarify the role of ANXA1 in OSCC tumorigenesis,through intervention of ANXA1 expression in OSCC cell lines.Another aim is to explore the relationship of ANXA1 expression and the sensitivity of oral cancer cell lines to the chemoagents of docetaxel,cisplatin,and 5-fluorouracil,in order to clarify whether ANXA1 expression could be used as a predictive biomarker for TPF induction chemotherapy.Methods:1.The role of ANXA1 on the proliferation and cell cycle in OSCC lines was detected by siRNA interference and gene cloning technology;2.Western blot analysis was used to test the influence of ANXA1 level on EGFR/PI3 K/Akt signaling pathway transduction.3.CCK8 assay was used to detect the sensitivity of OSCC cells to docetaxel,cisplatin and 5-fluorouracil drugs with ANXA1 interference.Western blot analysis was used to detect the apoptosis proteins of cleaved PARP and cleaved casepase-3.The results were analyzed using the software of Graphpad Prism 5.0,all hypothesis-generating tests were 2-sided at a significant level of P value less than 0.05.Results1.ANXA1 downregulation increased the cell proliferation in the HB96,HN4 cells as well as the percentage of S phase in the cell cycle;ANXA1 overexpression inhibited the CAL27 cell proliferation and the percentage of S phase in the cell cycle;2.In the HB96 and HN4 cells with downregulation of ANXA1,increased phosphorylation of EGFR and Akt,PKD1,c-Raf,which were associated with the EGFR/PI3K/Akt signaling pathway,and decreased level of p27kip was also found in these cells.On the contrary,in the CAL27 cells with ANXA1 overexpression,decreased phosphorylation of EGFR,Akt,PKD1,and c-Raf were found with increased p27kip expression;3.Compared with the control,the HN4 and HB96 cells were more sensitive to TPF drugs,and the expression of cleaved PARP and cleaved caspase-3 was significantly up-regulated with downexpression of ANXA1.Compared with the control,the CAL27 cells was less sensitive to TPF drugs,and the expression of cleaved PARP and cleaved caspase-3 was significantly decreased with overexpression of ANXA1.Conclusions:1.ANXA1,like a tumor suppressor gene,inhibited the transduction of EGFR/PI3K/Akt signaling pathway to supress the cell cycle and cell proliferation;2.The expression level of ANXA1 was related to the sensitivity of oral cancer cell lines to TPF drugs by influenced the ability of chemoreagent to induce the oral cancer cell apoptosis;3.ANXA1 might be used as a potential biomarker to predict the prognosis of OSCC patients,who could benefit from TPF induction chemotherapy.