[Effects of Osthole on Apoptosis and Autophagy of Human Tongue Cancer Tca8113 Cells].

Objective To investigate the effects of osthole on the proliferation,apoptosis,and autophagy of human tongue cancer Tca8113 cells and explore its possible mechanism of action. Methods Tca8113 cells were cultured in vitro and divided into a control group without drugs and the experimental groups with 40,80,120,and 160 µmol/L osthole.The inhibitory effect of osthole on the proliferation of Tca8113 cells was detected by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay and colony formation assay.Hoechst33342 staining method and annexin V-FITC/propidium iodide method were employed to detect the effect of osthole on the apoptosis of Tca8113 cells within 24 hours.Western blot was performed to detect the expression of apoptosis-related proteins(Bcl-2,Bax,and cleaved caspase-3)and autophagy-related proteins(LC3 and p62)in Tca8113 cells exposed to osthole. Results Osthole significantly inhibited the proliferation and induced the apoptosis of Tca8113 cells in a concentration-dependent manner,and it reduced the cell colony formation.Western blot results showed that osthole could up-regulate the expression of Bax and cleaved caspase-3 and down-regulate that of Bcl-2.At the same time,it increased the expression of LC3Ⅱ and P62 and reduced that of LC3Ⅰ. Conclusion Osthole may inhibit the proliferation of Tca8113 cells by promoting cell apoptosis and blocking autophagy flow to inhibit autophagy.

Apigenin Inhibits Proliferation, Migration, and Invasion of Human Tongue Carcinoma Tca8113 Cells Through Regulating the MAPK Signaling Pathways.

OBJECTIVE: This study aimed to examine the effects of apigenin (API) on the proliferation, migration, and invasion of human tongue squamous cell carcinoma Tca8113 cells and explore its probable mechanisms. METHODS: After treating Tca8113 cells with API, the cell proliferation, migration, and invasive capacities were identified by tetrazolium salt colorimetry (MTT) assay, cell scratch test, and Transwell chamber test. Cellular immunofluorescence staining was used to localize mitogen-activated protein kinase 1 (MAPK1) and extracellular regulated protein kinase (ERK) 1/2 proteins. Western blot was used to detect the variations of the related protein expression levels. RESULTS: 1)Through the MTT assay, API significantly inhibited cell proliferation (P<0.01). 2) In the cell scratch test, the distance of lateral migration after the API treatment was significantly shorter compared to the control group (P<0.01). 3) The invasion rate in the lower chamber of the Transwell chamber was lower in the API group (P<0.01). 4) Cellular immunofluorescence staining presented that the total-MEKK1 was localized in the cytoplasm, p-MEKK1 was localized in the nuclear membrane and cytoplasm, and p-ERK1/2 was localized in the cytoplasm and nucleus. ⑤ After API was applied to cells, the expressions of p-MEKK1 and p-ERK1/2 proteins significantly reduced (P<0.01). CONCLUSION: Apigenin (API) significantly inhibits the proliferation, migration, and invasion of Tca8113 cells and its mechanism may be associated with the MAPK signaling pathway.

Expression of miR-126 in oral squamous cell carcinoma tissues and its clinical significance / 中国肿瘤生物治疗杂志

@#[Abstract] Objective: To detect the expression of miR-126 in oral squamous cell carcinoma (OSCC) and to analyze its correlation with clinicopathological features and prognosis of patients, as well as to explore the effect of miR-126 over-expression on the malignant biological behaviors of Tca8113 cells. Methods: A total of 62 pairs of cancer and para-cancerous tissue specimens from OSCC patients who were surgically treated in the First Affiliated Hospital of Zhengzhou University from June 2016 to June 2018 were collected for this study; in addition, human tongue squamous carcinoma Tca8113 cell line and human mouth keratinocyte HOK cell line were also selected for this study. The expression of miR-126 in cancer tissues and cells was detected by qPCR, and the relationship between miR-126 expression and clinicopathological features and prognosis of the patients was analyzed. miR-126 mimics and miR-NC plasmids were respectively transfected into Tca8113 cells by liposome transfection technology. Cell proliferation, apoptosis, migration and invasion were detected by MTT method, Flow cytometry and Transwell chamber method, respectively; and the expressions of apoptosis, migration and invasion related proteins were detected by Western blotting. Results: The expression level of miR-126 in OSCC tissues and Tca8113 cells was significantly lower than that in para-cancerous tissues and HOK cells (all P<0.01). The expression of miR-126 was associated with TNM stage and lymph node metastasis (all P<0.05), and patients with high miR-126 expression had significantly better overall survival rate than patients with low expression (P<0.05). After transfection with miR-126 mimics, the cell proliferation, migration and invasion ability significantly decreased (P<0.05 or P<0.01) while the apoptosis rate significantly increased in Tca8113 cells (P<0.01), the expression levels of Bcl-2, N-cadherin and vimentin in Tca8113 cells significantly decreased (all P<0.01), and expression levels of Bax and E-cadherin significantly increased (all P<0.01). Conclusion: miR-126 is low expressed in OSCC tissues and Tca8113 cells. Up-regulation of miR-126 inhibits cell proliferation, migration and invasion and promotes apoptosis of Tca8113 cells.

Effects of Osthole on Apoptosis and Autophagy of Human Tongue Cancer Tca8113 Cells / 中国医学科学院学报

Objective To investigate the effects of osthole on the proliferation,apoptosis,and autophagy of human tongue cancer Tca8113 cells and explore its possible mechanism of action. Methods Tca8113 cells were cultured

Study on mechanism of down-regulating ikca1 molecule affecting the increment of oral squamous cell carcinoma.

This research aimed to explore the mechanism of mediating the down-regulation of the calcium-activated potassium channel (IKCa1) gene expression in human oral squamous cell carcinoma Tca-8113 cells, thereby affecting cell proliferation and apoptosis. The expression level of IKCa1 in Tca-8113 cell line (oral squamous cell carcinoma) and HOEC cell line (human normal oral epithelial cell) was detected by RT-PCR. Then, after IKCa1 was knocked down in Tca-8113 cell line and HOEC cell line by RNA interference, and then cell proliferation levels were detected by cell counting kit 8 (CCK-8) method. Cell cycle distribution was detected by flow cytometry. Apoptosis was detected by membrane linked protein V-FITC/propidium iodide (PI) double-staining apoptosis detection kit. The protein expression level of IKCa1 was detected by Western Blot method. According to RT-PRC results, IKCa1 was significantly more expressed in Tca-8113 cell line than in HOEC cell line (P< 0.01).  In addition, the mRNA expression levels in the normal oral epithelium and oral squamous cell carcinoma showed the same trend. After knocking down IKCa1 in Tca-8113 cell line, the IKCa1siRNA group significantly inhibited cell proliferation compared with the siNC control group. The results of flow cytometry showed that the proportion of apoptotic Tca-8113 cells transfected with IKCa1siRNA was significantly increased. The ratio of early apoptosis and late apoptosis of Tca-8113 cells increased (P< 0.05). To investigate the effect of IKCa1 on apoptosis, we tested the expression levels of apoptosis-related proteins. The results showed that the mRNA level of IKCa1siRNA group was significantly decreased by 44.41% compared with the control group (p< 0.01). Meanwhile, the mRNA level of Bax was significantly increased by 36.0% (p< 0.05). Our results showed that knocking down IKCa1 in Tca-8113 cells could induce cell cycle arrest and apoptosis to produce an anti-proliferation effect, thus inhibiting the expression of IKCa1 has an anti-cancer effect in oral squamous cell carcinoma.

Identification, characterization and microRNA expression profiling of side population cells in human oral squamous cell carcinoma Tca8113 cell lines.

The present study aimed to evaluate the stem cell markers, characteristics and biological functions of cancer stem­like side population (SP) cells in human oral cancer. SP cells were isolated from the human oral squamous cell carcinoma Tca8113 cell line by Hoechst 33342 fluorescence dye and flow cytometry. The colony forming and proliferative capability of SP and non­SP cells were detected using a live­cell analysis system in vitro. The number of cells expressing stem cell markers was compared between SP cells and non­SP cells by flow cytometry. Reverse transcription­quantitative polymerase chain reaction and western blotting were used to detect the mRNA and protein expression levels of stem cell genes, respectively. Differential expression of microRNAs (miRNAs) in SP and non­SP cells was determined by microarray hybridization and an miRNA regulation network was produced. With regard to the proliferation capability, SP cells reached 60.0% confluence after 40 h of growth compared with 35.1% confluence for non­SP cells (P<0.05). The number of colonies in SP cells was 43.1±9.2 compared with 33.0±8.2 of non­SP cells (P<0.05). The aldehyde dehydrogenase­1 (ALDH1)­positive cell number in the SP cells was increased by 10 times compared with the non­SP cells (P<0.01). The mRNA and protein expression levels of ALDH1, SRY­box 2, POU class 5 homeobox 1 and Nanog homeobox in SP cells were significantly higher compared with non­SP cells (P<0.05). Microarray hybridization demonstrated that 21 miRNAs were upregulated and 13 miRNAs were downregulated in SP cells compared with non­SP cells. SP cells in Tca8113 demonstrated greater capability of proliferation and colony formation compared with non­SP cells in vitro. Stem cell markers were overexpressed in SP cells compared with non­SP cells.

Apoptosis of Tca8113 cell induced by antitumor component-I from Agkistrodon acutus venom through CHOP/Caspase-12 pathway / 中国临床药理学与治疗学

AIM: To investigate the effect of endoplasmic reticulum stress pathway on the apoptosis of tongue squamous cancer Tca8113 cells induced by antitumor component-I from Agkistrodon acutus venom (AAVC-I). METHODS: The in vitro experiments were performed on subculture tongue squamous cancer Tca8113 cells in their growth period. A normal control group, a DL-dithiothreitol (DTT) positive control group and different AAVC-I concentrations were set according to the experiment objective. MTT assay was used to detect the proliferation inhibition of Tca8113 cells after been treated with different concentrations of DTT and AAVC-I for 24 h. The results were used to choose appropriate concentrations of DTT and AAVC-I in DTT positive control group and AAVC-I treated group, respectively. HE staining and Annexin V-FITC/PI double fluorescence staining were used to monitor the apoptosis of Tca8113 cells. Western blot was used to identify the expression levels of apoptosis-related proteins including endoplasmic reticulum stress glucose-regulatory protein 78 (GRP78), enhance-binding protein-homologousprotein (CHOP), cysteine-containing aspartate specific protease-12 (Caspase-12), cysteine-containing aspartate specific protease-9 (Caspase-9) and cysteine-containing aspartate specific protease-3 (Caspase-3).RESULTS:The proliferation inhibition of Tca8113 cells increased with an increased concentration of AAVC-I concentration (P<0.05), causing cell shrinkage, increased cell gaps, cytonuclear condensation, cell fragmentation, the appearance of apoptotic bodies, and increased rate of apoptosis (P<0.05). In addition, the expression level of GRP78 protein, CHOP protein, proteins of Caspase-12, Caspase-9 and Caspase-3 were increased (P<0.05).CONCLUSION: Endoplasmic reticulum stress CHOP/Caspase-12 pathway plays an important role in AAVC-I induced Tca8113 cells apoptosis.

Effect of silencing of mediator of DNA damage checkpoint protein 1 on the growth of oral squamous cell carcinoma in vitro and in vivo.

Mediator of DNA damage checkpoint protein 1 (MDC1) is involved in DNA damage repair and has been linked to tumor invasion, metastasis, and prognosis. This study investigated the effects of MDC1 in oral squamous cell carcinoma (OSCC) in vitro and in vivo. RNA interference-mediated knockdown of MDC1 was performed in two OSCC cell lines (Tca-8113 and KB). Real-time PCR and western blotting were performed to determine expression of mRNA and protein, respectively, of MDC1. Cell viability was assessed using the MTT assay. Colony-formation assays were performed by staining with 0.5% crystal violet. Cell migration and invasion were detected by Transwell assays. The role of MDC1 in OSCC was examined in vivo via injection of Tca-8113 cells transfected with MDC1 small interfering (si)RNA or negative-control siRNA into a mouse xenograft model of OSCC. Our results showed that MDC1 knockdown decreased cell proliferation. Inhibition of MDC1 decreased colony formation of Tca-8113 and KB cells by 62% and 68%, respectively, and MDC1 knockdown reduced the number of migratory and invasive cells compared with the control group. Moreover, the xenograft mouse model of MDC1 knockdown showed reduced tumor growth. Our study suggests that MDC1 plays a role in tumorigenesis and might be a potential target for the treatment of patients with OSCC.

Inhibitory effect of asiatic acid on proliferation and apoptosis of TCA-8113 human tongue carel inoma cells / 中国药理学通报

Aim To investigate the effect of asiatic acid(AA) on the proliferation, apoptosis and cell cycle arrest of human tongue carcinoma TCA-8113 cells, and the possible mechanism. Methods The inhibitory effect of AA(0, 20, 30, 40, 50 pjnol • L"1) at different concentrations on TCA-8113 cells for 24 h was determined by MTT. The effect of AA on the colony formation rate of TCA-8113 cells was detected by colony formation test; The apoptosis of TCA-8113 cells by AA was detected by Hoechst 33342 staining and Annexin V-FITC/PI staining. Cell cycle changes were analyzed by flow cytometry. The expressions of protein in human tongue cancer cells, such as bcl-2, Bax, cleaved caspase-3, p53 and p21 proteins were detected by Western blot. Results The inhibitory rate of AA on TCA-8113 cells was concentration-dependent (P < 0.05), with the IC50 concentration of 42. 13 (xmol • L- 1, and the ability of cell colony formation decreased. The apoptosis rate increased with the increase of AA concentration (P < 0. 05). The concentration of AA 30 u.mol -L"1 gradually blocked the cell cycle in G2/M phase. Western blot analysis showed that the expressions of p53, p21 and Bax increased, while Bcl-2 decreased in a dose-dependent effect (P<0. 05). Conclusions AA can significantly inhibit the proliferation and apoptosis of human tongue carcinoma TCA-8113 cells, and its mechanism may be related to the regulation of p53 pathway related proteins p53, p21, Bax and the inhibition of Bcl-2 expression, and AA can arrest TCA-8113 cells cycle in G2/M phase.

lncRNA XIST/miR-34a-5p/SIRT6 axis regulates proliferation and metastasis of oral squamous cell carcinoma cells / 中国肿瘤生物治疗杂志

@# Objective: To investigate the molecular mechanisms of lncRNA XIST/miR-34a-5p/SIRT6 axis regulating the proliferation and metastasis of oral squamous cell carcinoma (OSCC) cells. Methods: Specimens of tumor tissues and paracancer tissues of 47 patients with OSCC, who visited the Qingdao Stomatological Hospital from March 2013 to March 2018, were enrolled in this study. qPCR was performed to measure the mRNA expressions of lncRNA XIST, miR-34a-5p and SIRT6 in OSCC tissues and cell lines. WB was performed to measure the protein levels of SIRT6, Ki67, pcDNA, cleaved-caspase3, cleaved-caspase8, E-cadherin and vimentin in OSCC tissues and cell lines. CCK-8 assay was performed to observe the effect of lncRNA XIST knockdown on proliferation of Cal-27 and Tca-8113 cells; Tanswell assay was performed to detect migration and invasion of Cal-27 and Tca-8113 cells; flow cytometry was performed to detect the apoptosis of Cal-27 and Tca-8113 cells; and dual luciferase reporter assay was performed to verify the relationship between lncRNAXIST and miR-34a, or miR-34a and SIRT6. Results: Expressions of lncRNAXIST and SIRT6 were up-regulated in OSCC tissues and cell lines (all P<0.05), reversely, miR-34a-5p was down-regulated in OSCC tissues and cell lines (P<0.01). lncRNA XIST knockdown significantly suppressed OSCC cells proliferation, migration and invasion, and induced apoptosis of OSCC cells (all P<0.01); however, simultaneous transfection with miR-34a-5p inhibitor or pcDNA-SIRT6 vector exerted opposite effect. lncRNA XIST knockdown significantly increased cell proliferation and metastasis related protein expression in OSCC cells (all P< 0.01), and simultaneous transfection with miR-34a-5p inhibitor or pcDNA-SIRT6 vector exerted opposite effect. In addition, lncRNA XIST directly targeted miR-34a-5p, and miR-34a-5p targeted SIRT6, lncRNA XIST upregulated SIRT6 expression via inhibiting miR34a-5p (P<0.01). Conclusion: Taken together, lncRNA XIST/miR-34a-5p/SIRT6 axis regulates the proliferation and metastasis of OSCC cells and provides potential therapeutic targets for OSCC.

Effect of TET1 regulating MGMT on chemotherapy resistance of oral squamous cell carcinoma stem cells.

The study was to evaluate the effect of ten-eleven translocation 1 (TET1) regulating o6-methylguanine-DNA methyltransferase (MGMT) in chemotherapy resistance of oral squamous cell carcinoma (OSCC) stem cells. OSCC stem cells were divided into the blank, negative control (NC), TET1-siRNA, TET1-siRNA + MGMT-OE, and MGMT-OE groups. Methylation-specific polymerase chain reaction (MSP), qRT-PCR and Western blotting were conducted to detect the methylation status of MGMT, expressions of TET1, MGMT, ABCG2, and Oct-4. Cell proliferation, cisplatin chemosensitivity, and cell cycle and apoptosis, were detected using CCK8 and flow cytometry. A chromatin immunoprecipitation (ChIP) assay was employed for detecting the link between TET1 and MGMT gene promoters. In comparison to the NC group, the TET1-siRNA group exhibited increased levels of MGMT methylation, the number of apoptotic cells and cisplatin chemosensitivity consisting of varying concentrations, however, decreased levels of mRNA and protein expressions of TET1 as well as MGMT, cell viability, the number of cells in the S phase, and protein expressions of ABCG2 and Oct-4 were all have diminished amounts. The TET1-siRNA + MGMT-OE and MGMT-OE groups had higher MGMT mRNA and protein expression, as well as increased protein expressions of ABCG2 and Oct-4, greater cell activity, higher number of cells in the S phase, decreased apoptotic rates in cells and decreased cisplatin chemosensitivity with different concentrations. Our study provided evidence that low-expression of TET1 in OSCC stem cells may stimulate MGMT promoter methylation, while inhibiting MGMT mRNA expression, this ultimately strengthens the sensitivity of OSCC stem cells in regards to chemotherapeutics.

Analysis on gene profile of over-expression WWOX gene in human oral squamous cell carcinoma Tca8113 cells and tumor inhibition mechanism of WWOX gene / 吉林大学学报(医学版)

Objective: To analyze the changes of gene profile of over-expression WWOX gene in the human oral squamous cell carcinoma (OSCC) Tca8113 cells, and to explore the tumor inhibition mechanism of WWOX gene. Methods: The Tca8113 cells were divided into WWOX over-expression group and control group. In WWOX over-expression group, the lentiviral plasmid carrying the full-length cDNA fragment of WWOX gene was infected into the Tca8113 cells, while in control group, the lentiviral plasmid carrying the empty pGV287-LV vector was infected into the Tca8113 cells. The quantitative real time PCR (qRT-PCR) and Western blotting methods were used to detect the expressions of WWOX mRNA and protein. The gene chip technique was used to screen the differientially expressed genes and biological analysis was performed. The quantitative PCR was used to detect the relative mRNA expression levels of differientially expressed genes involving in the mitogen-activated protein kinase (MAPK) signaling pathway. Results: After infection, the relative mRNA expression level of WWOX gene in the Tca8113 cells in WWOX over-expression group was higher than that in control group (P<0. 05), and the expression of WWOX protein in the Tca8113 cells in WWOX over-expression group was higher. A total of 347 differientially expressed genes with a 1. 5-fold-change (FC) were screened by gene chip, in which 171 genes showed up-regulated expression, and 176 genes showed down-regulated expression. The Bioinformation analysis results showed that these different genes distributed in several pathways, which were related to cancer, p53, MAPK, and interleukin-2 (IL-2) pathways and so on. Compared with control group, the relative expression levels of MAP2K, NR4A1, DUSP5, and DUSP6 mRNA in MAPK signaling pathway in WWOX over-expression group were increased (P<0. 05), and the relative expression level of fibroblast growth factor receptor 2 (FGFR2) mRNA was decresed (P<0. 05). Conclusion: The over-expression of WWOX gene in the Tca8113 cells can induce the changes of gene profile of Tca8113 cells. The tumor inhibition mechanism of WWOX gene may be related to regulating the expressions of some genes involving in MAPK signaling pathway.

Inhibitory effect and mechanism of Nultin-3 combined with cisplatin on oral squamous cell carcinoma / 实用肿瘤学杂志

Objective The objective of this study was to investigate the inhibitory effect of MDM2 inhibitor Nultin-3 com-bined with cisplatin on human oral squamous cell carcinoma(OSCC)Tca8113 cells and its mechanism. Methods Human OSCC Tca8113 cells were treated with Nultin-3,cisplatin,Nultin-3 combined with cisplatin,or vehicle control groups. The proliferation of Tca8113 cells was determined by thiazolyl blue(MTT)assay. The expression of MDM2,P53,Caspase-9 and Caspase-3 protein was determined by Western blot. Results The proliferative rate of OSCC ca8133 cells treated with Nultin-3 combined with cisplatin was significantly lower than that of other groups(P<0. 05). The relative expression of Caspase-9,Caspase-3 and P53 protein in the Nultin-3 combined with cisplatin was significantly higher than those of the Nultin-3 and cisplatin alone groups(P<0. 05). In addi-tion,the relative expression of MDM2 protein in the Nultin-3 combined with cisplatin group was significantly lower than that of the cisplatin and Nultin-3 alone groups(P<0. 05). Conclusion Nultin-3 combined with cisplatin has synergistic effect on oral squa-mous cell carcinoma Tca8113 cells. Nultin-3 regulates the MDM2-p53 signaling pathway and up-regulates the expression of pro-apoptotic proteins Caspase-9 and Casapase-3 to enhance the inhibition of cisplatin on oral squamous cell carcinoma,providing a solid theoretical basis for clinical research and treatment.

Stromal interaction molecule 1 regulates growth, cell cycle, and apoptosis of human tongue squamous carcinoma cells.

Oral tongue squamous cell carcinoma (OTSCC) is the most common type of oral carcinomas. However, the molecular mechanism by which OTSCC developed is not fully identified. Stromal interaction molecule 1 (STIM1) is a transmembrane protein, mainly located in the endoplasmic reticulum (ER). STIM1 is involved in several types of cancers. Here, we report that STIM1 contributes to the development of human OTSCC. We knocked down STIM1 in OTSCC cell line Tca-8113 with lentivirus-mediated shRNA and found that STIM1 knockdown repressed the proliferation of Tca-8113 cells. In addition, we also showed that STIM1 deficiency reduced colony number of Tca-8113 cells. Knockdown of STIM1 repressed cells to enter M phase of cell cycle and induced cellular apoptosis. Furthermore, we performed microarray and bioinformatics analysis and found that STIM1 was associated with p53 and MAPK pathways, which may contribute to the effects of STIM1 on cell growth, cell cycle, and apoptosis. Finally, we confirmed that STIM1 controlled the expression of MDM2, cyclin-dependent kinase 4 (CDK4), and growth arrest and DNA damage inducible α (GADD45A) in OTSCC cells. In conclusion, we provide evidence that STIM1 contributes to the development of OTSCC partially through regulating p53 and MAPK pathways to promote cell cycle and survival.

Effects of PTEN gene silencing on invasion and EMT in oral squamous carcinoma Tca8113 cells.

AIM: To investigate the impact of silencing of the PTEN gene using siRNA on the invasion, proliferation, cell cycle, and epithelial-mesenchymal transition of the Tca8113 cell line. METHODS: The established Tca8113 cell model with siRNA interference to silence the PTEN gene was used. The transfection efficiency was examined by RT-qPCR and Western blot analysis. CCK-8 assay was utilized to analyze the proliferation of Tca8113 cells and cell invasion was evaluated using a transwell assay. The cell cycle distribution was analyzed by flow cytometry. The protein expression levels of the epithelial-mesenchymal transition (EMT) markers E-cadherin and Vimentin and the EMT-related proteins ß-catenin and TGF-ß1 were analyzed by Western blot. RESULTS: The expression level of PTEN was significantly reduced in the PTEN-siRNA group. The invasiveness and proliferation rate of Tca8113 cells in the PTEN-siRNA group were significantly greater than those of the control and negative control groups. The expression levels of E-cadherin and ß-catenin were reduced, whereas the expression levels of vimentin and TGFß-1 were elevated in the PTEN-siRNA group compared with those of control and negative groups. These results were significantly different. CONCLUSION: The silencing of PTEN by siRNA increased the proliferation and promoted cell invasion of Tca8113 cells. PTEN gene silencing may accelerate the EMT in Tca8113 cells.

Contrastive Analysis of three transplanted tumor models in chick embryo chorioallantoic membrane / 中国比较医学杂志

Objective To compare the growth of three different cancer cell lines on chick chorioallantoic membrane (CAM), to select the best transplanted cancer cell line for establishing a transplanted tumor model and to observe the biological characteristics.Methods The human lung cancer cell line A549, human tongue cancer cell line TCA8113 and human liver cancer cell line QGY7703 were respectively inoculated into CAM at the 7th day of age.The chick embryo survival rate, tumor survival rate, tumor formation rate and induced angiogenesis were detected and the growth characteristics of the transplanted tumor model were observed.Results Compared with the groups inoculated with A549 cells and QGY7703 cells, the tumor formation rate of TCA8113 cells was the highest (P < 0.05), to be the best cancer cell line for transplanted tumor.The optimal inoculated number of cells was 8.0×106/chick embryo, the optimal growth period of the tumor was 4~8 d, and the best experiment time was 7 d after inoculation.Conclusion The TCA-CAM transplanted tumor model of tongue squamous cell cancer is successfully established for further study of the biological characteristics and mechanisms of tumor growth, angiogenesis, invasion and metastasis, and provide a good experimental animal model for anti-tumor drug screening.

Sorafenib inhibits the proliferation of human oral cancer TCA8113 cells through the activation of p38MAPK / 实用口腔医学杂志

Objective:To investigate the effect of sorafenib on the proliferation of human oral cancer TCA8113 cells and to explore the underlying mechanisms.Methods:Mter treated with sorafenib at 2.5,5,10,20 μg/ml respectively for48 h,TCA8113 cell proliferation was examined by MTT and colony formation assay.Western blotting was employed to examine the p38MAPK expression in the cells.TCA8113 cells were pretreated with 10 μmol/L of SB203580 (a specific inhibitor of p38MAPK) for 30 min,and then by different concentrations of sorafenib for 48 h,cell proliferation was tested by MTT assay.Results:Sorafenib significantly inhibited the proliferation of TCA8113 cells in a concentration dependent fashion.Sorafenib also remarkably promoted the activation of p38MAPK of the cells.SB203580 significantly alleviated soiafenib induced TCA8113 cell viability decrease.Conclusion:Sorafenib can inhibit the proliferation of TCA8113 cells,which may be related to the activation of p38MAPK.

Effect of Curcumin Combined with Cisplatin on Human Tongue Squamous Cell Carcinoma Tca?8113 Cells / 中国医科大学学报

Objective To observe the effect of curcumin combined with cisplatin on the proliferation of human tongue squamous cell carcinoma Tca?8113 cells,and explore the possible mechanism of their anti?tumor effect in vitro. Methods In this study,the samples were divided into four groups:curcumin,cisplatin,curcumin combined with cisplatin,and control. Detection of rate of inhibition of Tca?8113 cell proliferation was carried out by MTT assay. This was followed by observation of the morphological changes of the nuclei by Hoechst 33258 fluorescence staining. Subse?quently,cellular apoptosis was assayed by flow cytometry,and expression of Notch1 and epidermal growth factor receptor(EGFR)was examined by Western blotting. Results Results of MTT assay showed that curcumin combined with cisplatin inhibited the proliferation of tongue squamous cell carcinoma to a greater extent than either curcumin or cisplatin alone(P<0.05). Flow cytometry analysis indicated that,unlike curcumin or cisplatin alone,curcumin combined with cisplatin noticeably induced apoptosis(P<0.05). Results of Western blotting revealed that the expres?sion of Notch1 was upregulated,whereas the expression of EGFR was downregulated to a greater extent in the control group than in cells treated with curcumin or cisplatin alone. Unlike curcumin and cisplatin groups,the combined group revealed statistically significant expressions of Notch1 and EGFR(P<0.05). Conclusion Curcumin and cisplatin have a combined effect on inhibition of proliferation of human tongue squamous cell carcinoma Tca?8113 cells. The mechanism may be related to upregulation of the Notch1 signaling pathway and downregulation of the EGFR signal?ing pathway.