Clones Morphological Observation Of Adriamycin-induced Human Tongue Cancer Drug-resistant Cell And Stuyd On The Biological Characteristics Of Clones

Objective In order to enrich the high proportion of tongue cancer stem cell-like cells,to detect the biological characteristics of adriamlycin-induced human tongue cancer drug-resistant cell clones based on the. resistance characteristics of cancer stem cells and unicellular culture.Methods In the first part:using unicellular culture for two weeks,Tca-8113/ADM-P4 and drug survival cells by 0.01?g/ml,0.05?g/ml adriamycin intervention 48 hours (named as 0.01?g/mlDSC,0.05?g/mlDSC)would be observed for the biological characteristics of clones, including the clone formation rate, clone constituent ratio and clone growth curve. In the second part:The holoclone cells and meroclone cells of Tca-8113/ADM-P4,0.01?g/mlDSC and 0.05?g/mlDSC would be continuously sub-cultured in vitro and then brought into immunodeficiency tumor formation in nude mice and tested by quantitative real-time PCR in order to detect clonal characteristics.Results The results of the first part : (1)Using unicellular culture,Tca-8113/ADM-P4,0.01?g/mlDSC and 0.05?g/mlDSC cells can generate holoclone,meroclone and paraclone cells.The total clone formation rate of Tca-8113/ADM-P4 cells was 70.75%,whice the constituent ratios of holoclone,meroclone and paraclone cells were 29.05%,48.60% and 22.35%.The total clone formation rate of 0.01?g/mlDSC cells was 58.27%,whice the constituent ratios of holoclone,meroclone and paraclone cells were 19.75%,27.16% and 53.09%.The total clone formation rate of 0.05?g/mlDSC cells was 58.27%,whice the constituent ratios of holoclone,meroclone and paraclone cells were 13.92%,31.65% and 54.43%.(2)The growth curve result of Tca-8113/ADM-P4,0.01 ?g/mlDSC and 0.05?g/mlDSC clone cells:the proliferation rate of holoclone cells was the fastest,and the pareclone cells was the slowest,the proliferation rate of the meroclone cells was between the two.The results of the second part:(1)Holoclone and meroclone cells of Tca-8113/ADM-P4,0.01 ?g/mlDSC and 0.05?g/mlDSC would be continuously sub-cultured in vitro.(2)The results of quantitative real-time PCR showed that the expression of ABCG2,CXCR4 in 0.05?g/mlDSC holoclone and meroclone cells were higher than Tca-8113/ADM-P4 and 0.01?g/mlDSC,the difference was statistically significant(P<0.05).(3) Holoclone cells of Tca-8113/ADM-P4,0.01?g/mlDSC and 0.05?g/mlDSC were able to form tumors in the nude mice,and the tumor formation rate was 100%. The time of tumor formation of 0.05?g/mlDSC holoclone cells was shortened than Tca-8113/ADM-P4 holoclone cells,the difference was statistically significant(P<0.01);Meroclone cells of Tca-8113/ADM-P4, 0.01?g/mlDSC and 0.05?g/mlDSC were able to form tumors in the nude mice, and the tumor formation rate was 100%,97.22%,94.44%. The time of tumor formation of 0.05?g/mlDSC meroclone cells was shortened than Tca-8113/ADM-P4 meroclone cells, the difference was statistically significant(P<0.05).Conclusions (1)Using unicellular culture,Tca-8113/ADM-P40.01?g/mlDSC and 0.05?g/mlDSC cells can form holoclone,meroclone and pareclone cells. (2)Meroclone cells of Tca-8113/ADM-P4 can be continuouslysub-cultured in vitro to form holoclone cells,it may contain cancer stem like cells,whice have self-renewal,proliferation and tumorigenicity characteristics.(3)Based on the resistance characteristics of cancer stem cells and unicellular culture,Tca-8113/ADM-P4 after adriamycin intervention can further enrich cancer stem cell-like cells.