Detection Of Giant Cancer Cells In Human Oral Squamous Cell Carcinoma And Their Correlation With Clinicopathological Characteristics And Mechanism

Objective This study aims to detect giant cancer cells in human oral squamous cell carcinoma(OSCC)and elucidate their correlation with OSCC clinicopathological characteristics.And this study also aims to investigate the possible mechanism of the induction or purification of GCC and explore the role of GCC on cell proliferation,chemoresistance and migration in vitro.Methods 1.Tumor paraffin sections and clinicopathological information of 76 OSCC cases were collected from Tianjin Medical University Cancer Institute and Hospital.Morphological differences between giant cancer cells and normal-sized cancer cells were observed under H&E staining and Ki-67 immunochemistry staining,respectively.The number of giant cancer cells was counted under a microscope(×400).The association between the number of giant cancer cells and the biological behavior of OSCC was analyzed.2.The concentration of Co Cl2 added to OSCC cell line UM1 was decided by CCK-8 assay.The expression of stemness markers were detected by real-time PCR and western blotting.The effect of GCCs on cell proliferation,chemoresistance and migration abilities were detected by CCK-8 and wound-scratch assays.The expression of HIF-1?and EMT-related markers were detected by western blotting.Results 1.Giant cancer cells were observed in OSCC tissues,and these cells were positive for Ki-67 staining.The number of giant cancer cells was closely associated with clinical stage(P=0.02),histological grade(P<0.01),chemotherapy(P=0.01),and local recurrence(P=0.02).By contrast,the number of giant cancer cells was not associated with sex,age,and lymph and distant metastases in OSCC(P>0.05).Patients with more than 3/HP giant cancer cells retained poor prognosis(P<0.05).2.UM1 can be induced to form GCCs by Co Cl2.After Co Cl2 treatment,almost all regular-sized cells were dead,and several purified GCCs survived and recovered.The size of the GCCs were bigger than regular cancer cell and some GCCs contained multiple nuclei.One main asymmetric cell division pattern was observed in GCCs: budding.Budding of daughter cells from GCCs usually occurred from the branches of GCCs.Compared with UM1,GCCs with budding cells upregulated mi RNA of stemness markers(NANOG and SOX-2).The expression of HIF-1 ?,SOX-2,Vimentin and N-cadherin were upregulated and the expression of E-cadherin was decreased in GCCs with budding cells.The proliferation,chemoresistance and migration abilities were upregulated in GCCs with budding cells.Conclusions 1.The number of giant cancer cells was closely associated with clinical stage,histological grade,chemotherapy,and local recurrence.Giant cancer cells may play an important role in the progression,chemoresistance,and prognosis of OSCC.2.OSCC cell lines can be induced and purified to cancer stem cell-like cells GCCs by Co Cl2.GCCs with budding cells may upregulated cancer cell proliferation,chemoresistance and migration abilities by EMT and upregulating stemness markers.