Objective To investigate the effects of miR-200c on proliferation and apoptosis of tongue squamous cell carcino-ma (TCCS)Tca8113 cells.Methods The mimics of miR-200c were transfected into Tca8113 cells using liposome.The Tca8113 cell proliferation was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT)assay.The flow cytometry as-say was used to determine the cell cycle and the apoptosis rate of Tca8113 cell.The protein expression levels of Bcl-2 and Caspase-3 in Tca8113 cell was detected by Western-blot.Results The 20,40,80 nmol/L miR-200c mimics groups inhibited the growth of Tca8113 cells,the difference compared with the control group showing statistical significance(P <0.05).The greater the miR-200c mimics concentration and the longer duration of action,the more significant the inhibition effect(P <0.05).After 48h transfecting by miR-200c mimics,the Tca8113 cells were arrested in the G0/G1 phases of cell cycle,and the apoptosis rate of the miR-200c mim-ics groups was significantly increased,the difference compared with the control group showing statistical significance(P <0.05);Western blot verified that the expression amount of Bcl-2 protein in the 20,40,80 nmol/L miR-200c groups was significantly lower than that in the control group,while the expression amount of Caspase-3 protein was significantly higher than that in the control group(P <0.05).Conclusion The overexpression of miR-200c might inhibit the proliferation of Tca8113 cell and induces their ap-optosis.

Objective To understand non stomatology undergraduates'strategies for learning stomatology and to study the reform on this course.Methods The learning strategies of 560 undergraduates majoring in clinical medicine from grade 2007 were investigated s and ten related factors like learning attitudes were investigated by learning strategies scale.Correlation analysis and linear regression analysis were applied to deal with research data.Results Most undergraduates were lack of strategies in learning stomatology.Related coefficient between 10 factors and academic scores ranged from 0.197 to 0.401,existing positive correlation(P＜0.05).Determination coefficients(R2)of attitude,motivation,time management and learning auxiliary means were 0.146,0.167,0.223and 0.122 respectively,which can be used to predict the scores of examination.Condusions Non stomatology undergraduates'strategies for learning stomatology is a vital factor influencing their academic scores.It's necessary for teachers to improve their teaching methods considering students'professional characteristics and learning strategies.

Oral and maxillofacial organoids (OMOs), tiny tissues and organs derived from stem cells cultured through 3-d cell culture models, can fully summarize the cell tissue structure, physiological functions and biological characteristics of the source tissues in the body. OMOs are applied in areas such as disease modelling, developmental and regenerative medicine, drug screening, personalized treatment, etc. Although the construction of organoids in various parts of the oral and maxillofacial (OM) region has achieved considerable success, the existing cocktail formulae (construction strategies) are not widely applicable for tissues of various sources due to factors including the heterogeneity of the source tissues and the dependence on laboratory technology. Most of their formulae are based on growth factor niches containing expensive recombinant proteins with their efficiency remaining to be improved. In view of this, the cocktail formulae of various parts of the OM organs are reviewed with further discussion of the application and prospects for those OMOs to find some affordable cocktail formula with strong operability and high repeatability for various maxillofacial organs. The results may help improve the efficiency of organoid construction in the laboratory and accelerate the pace of the clinical use of organoid technology.

OBJECTIVETo establish subseries cell lines from single, cancer cell of Tca8113M1 cell line and detect the cancer stem cell markers in the different subseries cell lines.

METHODSThe subseries cell lines from single cancer cell of Tca8113M1 cell line were established by limiting dilution assay in vitro. The characteristic of tumorigenicity and CD44, CD184, extracellular soluble antigen (ESA) of the cancer stem cell markers were detected by xenotransplantation and flow cytometry respectively.

RESULTSTotal 192 single cells of Tca8113M1 cell line were cultured and were deposited as one cell per well. There were 12 subpopulations origin from 192 single cells spheroid cultures. The ratio was 6.25% (12/192). In the different subpopulations, the tumorigenicity and expression of CD44 and ESA were at high levels, but the expression of CD184 was in different level. There were three kinds morphology of colonies derived from single cancer cells, holoclone, meroclone and paraclone. Cell line could be derived from carcinoma cell holoclones by cell culture. Meroclone and paraclone did not exist in cell culture in vitro.

CONCLUSIONTongue cancer stem cell may exist in Tca8113M1 cell line, cell line can be established and holoclone is the origin of cell line. This is a novel approach to the identification and enrichment for cancer stem cell.

Biomarkers, Tumor ; Cell Line ; Cell Line, Tumor ; Flow Cytometry ; Humans ; Neoplastic Stem Cells ; Tongue Neoplasms

Objective:To explore the role of casein kinase 1 gamma 2 (CSNK1G2) in the development and progression of head and neck squamous cell carcinoma (HNSC).Methods:Based on the Cancer Genome Atlas (TCGA), LinkedOmics and UALCAN were used to analyze the relationship among the mRNA expression of CSNK1G2, methylation, copy number variation and clinical indicators in HNSC, as well as to analysis CSNK1G2 related co-expression genes and proteins. The expression of CSNK1G2 in HNSC was verified by RT-qPCR experiments of clinical samples. Protein interaction network analysis on CSNK1G2 expression-related proteins was performed using STRING database.Results:UALCAN analysis showed that the expression of CSNK1G2 mRNA in HNSC was higher than that in normal tissues ( P<0.001), and the expression of CSNK1G2 mRNA was up-regulated in lower differentiation and Human Papilloma Virus (HPV)-positive HNSC (all P<0.05). But in HNSC with different pathological stages, different age stages and different lymph node metastasis stages (N stage), there was no difference in the amount of CSNK1G2 mRNA expression (all P>0.05). The RT-qPCR experiment confirmed the increased expression of CSNK1G2 mRNA in HNSC. LinkedOmincs analysis results showed that CSNK1G2 mRNA expression was positively correlated with CSNK1G2 copy number variation ( P<0.001) and negatively correlated with methylation ( P<0.001). Survival analysis results showed that high CSNK1G2 mRNA expression and copy number mutations predicted better survival ( P=0.033, P=0.015), while methylation levels were not associated with survival ( P=0.458). Gene set enrichment analysis results showed that CSNK1G2-related co-expression genes were mainly in DNA replication. The STRING's protein interaction network analysis results showed that TP53, CHEK1, and CHEK2 may be key proteins. These proteins are significantly associated with high expression levels of CSNK1G2. Conclusions:CSNK1G2 may cooperate with TP53, CHEK1 and CHEK2 related proteins to promote the development of HNSC and tumor proliferation, but does not affect the metastasis and spread of HNSC. An increase in the expression of CSNK1G2 in HNSC may indicate a better survival prognosis.