The Expressions Of Cancer Stem Cells Related Markers On Tongue Cancer Cells In Different Environments

Objecitve: To reasearch the expressions of cancer stem cells(CSCs) related markers on tongue cancer cells in different environments to provide more evidence of identification of CSCs markers of tongue cancer.Methods: To detect the expressions of cancer stem cell related markers(CD44, ESA, CXCR4, Nestin and Pan-CK) on tongue cancer cells(TCA8113) in standard culture medium (RPM1640+10%FBS) and"sphereround"culture medium(human Embryo stem cell basic midum +B27+bFGF+EGF ) with immunochemistry stainning and flow cytometry(FCM). And to inject TCA8113 cells into nude mice's subcutaneous of axillary fossa to establish subcutaneous transplanted tumor model; into foot pad of lower limb to establish lymphatic metastatic model and into peritoneal cavity to estabilish peritoneal cavity transplanted tumor model.then, inspecting the markers'expressions on tongue cancer cells.Results: Immunochemistry staining showed CD44, ESA, CXCR4, PAN-CK were highly expressed, yet nestin had low expresssion or negative on TCA8113 cells in standard culuture medium, FCM showed the frequencies are 98.2±0.49%(CD44), 99.7±0.21%(ESA) and 35.6±7.07%(CXCR4). After 22 days being cultured in"sphereround"culture medium, TCA8113 cells formed The"sphereround", the frequencies of"sphereround"cells with FCM were 99.9±0.03%(CD44), 98.7±1.19%(ESA) and 2.75±1.67%(CXCR4).After being cultured in standard culture medium, the morphology and growth pattern of "sphereround"turned like the cancer cells cultrued in standard culture medium, FCM showed the frequencies as fllows: 99.7±0.26 % (CD44), 99.0±1.14%(ESA), 36.7±4.63%(CXCR4) . The frequencys with FCM of lyTCA caner cells were 98.9±0.90% (CD44), 99.3±0.77%(ESA), 35.1±3.29%(CXCR4). All three models have been established successfully. The cancer cells in subcutaneous transplanted tumor highly expressed CD44, ESA, CXCR4, yet Pan-CK and nestin were lowly expressed. FCM indicated the frequencies were 84.3±5.17%(CD44), 94.0±3.22%(ESA), 85.6±6.37%(CXCR4). Immunoch- emistry staining showed CD44, ESA, CXCR4 and nestin were highly expressed in the cancer cells in metastatic lymph nodes of popliteal fossa, yet Pan-CK was lowly expressed. 1 month after cells injection into peritoneal cavity. A great many bloody ascites formed, FCM showed the frequencies of cancer cells were 95.3±4.59%(CD44), 99.9±0.05%(ESA), 79.4±3.93%(CXCR4).Conclusions: In different environments, CD44 and ESA are highly expressed on tongue cancer cells, yet the expressions of CXCR4, Pan-CK, Nestin vary.