Primary Research Of WNT7B Dependent Crosstalk Between OSCC Cells And Macrophages

Oral squamous cell carcinoma is among primary oral malignant tumors,and always be related with chronical oral inflammation.Although much progress in OSCC treatment has been developed recent years,the 5 years survival rate of OSCC is still low due to delayed primary site diagnosis,frequent local recurrence,and cervical lymph node metastasis after treatment.Our previous sequencing data of OLP and OSCC tissues from same patients revealed that,compared with the normal tissue,the expression of WNT7 B,a WNT ligand,was gradually increased in OLP and OSCC.To investigate the involvement of WNT7 B in development of OLP and OSCC,this project focus on the communicative mechanisms between tumor cells and tumor associated macrophages,purpose to disclose whether WNT7 B participate in the crosstalk between OSCC cells and TAMs.In the first part,we validated the involvement of WNT7 B in OLP and OSCC with clinical samples.Firstly,combined analysis of our previous transcriptome data and three patients' transcriptome data downloaded from GEO was performed,then followed by WNT7 B mRNA detection.The results showed that the expression of WNT7 B was gradually increased in normal tissue,OLP tissue and OSCC tissue.This result suggests that WNT7 B may play a role in the development of oral inflammation to cancer.The second part reveals that OSCC cells may take part in the activation of TAMs by secreated molecules including WNT7 B.M0 macrophages induced from THP1 cells by PMA was prepared first.And supernatant of cultured OSCC cells,as well as supernatant of WNT7 B stable knock down OSCC cells was collected to stimulate M0 macrophages,then checked the activation direction of M0 macrophages.The results indicated that the supernatant of OSCC cells can induce activation of M0 macrophages into classically activated M1,while supernatant from WNT7 B stable knockdown OSCC cells showed a reduced induction ability.The result suggests that WNT7 B,as a secreted protein,may be a M1 macrophages activation factor.In the third part,we validated that inflammatory factors secreated by M1 macrophages can affect the physiology characters of tumor cells.Supernatant of M1 macrophages derived from M0 macrophages under the induction of INF-? and LPS was harvested,and OSCC tumor cells were stimulated with above M1 macrophagesupernatant,then physiology characters of OSCC cells like proliferation and migration was checked,as well as expression of WNT7 B and other WNT signaling genes.WNT ligands are reported as secreted proteins.In our experiment,we found that the protein and mRNA expression level of WNT7 B was partly inconsistent.Other WNT ligands such as WNT3 have been reported to be released and influence other cells in an exosome dependent manner.We speculate that WNT7 B may also mediate cell-cell communication in a WNT3 similar pattern.This part of the work is still ongoing.