Intervening Effective Feature Of QI-boosting Toxin-Resolving Formula On Cellular Cycle Of CNE2NPC Cells To Inhibit Their Proliferating Activity And Its Association With The Expressive Activities Of Cancer Stem Cells Markers CD44and ABCG2

Objective To investigate the intervening effective feature of compound Chinese herbal medicine Qi-Boosting Toxin-Resolving Formula (QBTRF) on the cellular cycle of human nasopharyngeal carcinoma cell line CNE2to inhibit their proliferating activity and its association with the expressive activities of cancer stem cell markers CD44, ABCG2based on an experimental study.Methods1. Firstly done was to prepare the plasma containing the components of QBTRF (hebal medicine plasma) among rats, followed by determination on IC50concentation of hebal medicine plasma on target cells. Then, MTT assays were used to detect the inhibitory effects of QBTRF on the proliferating activity of CNE2cells,.2. The effective feature of QBTRF on cell cycle of CNE2cells were determined by flow cytometry with the most ideal effective concentration of hebal medicine plasma added into cultured target cells in vitro.3. At last, cell climbing slices of CNE2cells and Ha Ca cells were prepared for the intervening experiment of herbal medicine plasma on NPC cancer stem cells, with their expressive activities of biomarkers CD44and ABCG2detected by active cell immunohistochemistry, to explore the association of their expressive features with the intervening effect of QBTRF on the cell cycle and proliferation of CNE2.Results1. When compared with that of blank control group, the differences in cell proliferating activities of CNE2were statistically significant for those of herbal medicine plasma groups and cisplatin group (P<0.01). There were also statistical significance for differences in the proliferating activities among all the other groups (P<0.05) other than the group with10%herbal medicine plasma used, when compared with cisplatin group (P>0.05). It was shown in the experiment that the survival cells became less and less gradually along with the increase of medicine concentration in plasma, so as to show a concentration dependent feature. Moreover, the survival rate of CNE2cells was obviously lower for herbal medicine serum than that of herbal medicine plasma as compared at a same concentration level, but with no significantly statistical significance at the concentration of1%(P>0.05) between them.2. It was shown in the intervening experiment on cell cycle that the percentage of CNE2cells at G0/G1phase was elevated gradually with the increase of herbal medicine plasma concentration of QBTRF in a dose-dependent way when cultured with these cells for48hours as compared with that of blank control group (P<0.01). In contrast, the proportion of cells at S phase were markedly decreased (P<0.01) when compared with that of blank control group, in a dose dependent way also. However, there was no statistical significance in the proportion of cells at G2/M phase for groups intervened with herbal medicine plasma, though the percentage of cells at this phase decreased at a certain level, when compared with that of blank control group (P>0.05).3. The ratios of positive cells with CD44staining were0%,41.67%and25%for Ha Ca T group, CNE2group and herbal medicine plasma group respectively, while that of positive cells with ABCG2staining were2.13%,44.60%and32.78%for Ha Ca T group, CNE2group and herbal medicine plasma group respectively. There were statistical significance in their differences when compared among them (P<0.01).Conclusions1. Qi-Boosting Toxin-Resolving Formula holds significantly inhibitory effect on the proliferating activity of human nasopharyngeal carcinoma cell line CNE2, with such features as concentration-dependent and time-dependent effective ways.2. The intervening effect of Qi-Boosting Toxin-Resolving Fonnula on the proliferating activity of CNE2cells shows an obvious cell cycle specific feature leading to an arrest at GO/G1phase so as to promote these cells going into the progress of apoptosis rapidly.3. Qi-Boosting Toxin-Resolving Formula can give rise more significantly intervening effect on the expressive activities of cancer stem cell markers CD44and ABCG2on NPC cells CNE2. This kind of effect may be brought about by inhibiting cancer stem cells themselves. Then, arrest at GO/Gl phase will be taken place in cell population of CNE2, accompanying with an increase in apoptotic activity at the same time and a decrease in the proliferating ability among the cells further.