Proliferation Of Tca8113 Cells Research Articles

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.

Relationship Between mir15b and Sal-Like Protein 4 and Biological Behavior of Oral Squamous Cell Carcinoma

miR15b and SALL4 are involved in a variety of tumor progression. The roles of miR15b and SALL4 in oral squamous cell carcinoma (OSCC) remains unclear. The tumors and normal mucosa of OSCC patients were collected to detect miR15b and SALL4 level by Real-time PCR and analyze their correlation with OSCC clinicopathological features. Oral cancer Tca8113 cells were separated into control group; miR15b mimics group and miR15b inhibitor group followed by analysis of SALL4 expression, cell survival by MTT assay; cell invasion by Transwell chamber assay, as well as expression of N-cadherin and Vimentin and correlated with TNM stage, tumor volume and metastasis, and positively with differentiation TGF-β by Western blot. miR15b expression was decreased and SALL4 expression was increased in OSCC tumor tissues. miR15b was negatively degree (P < 0.05), whereas, opposite correlation of SALL4 with the above parameters was found (P < 0.05). miR15b and SALL4 were negatively correlated. MiR15b mimics significantly up-regulated MiR15b, decreased SALL4 expression, inhibited Tca8113 cell proliferation and invasion, as well as reduced N-cadherin, Vimentin and TGF-βexpression (P < 0.05). Opposite results were found in MiR15b inhibitor group. MiR15b expression is decreased and SALL 4 is increased in OSCC tumor tissues. MiR15b and SALL4 is closely related to OSCC clinicopathological features. MiR15b regulates the expression of EMT-related genes and TGF-β, thereby altering the proliferation and invasion of OSCC cells.

LncRNA HOXA-AS2 Promotes Oral Squamous Cell Proliferation, Migration, and Invasion via Upregulating EZH2 as an Oncogene.

Oral squamous cell carcinoma (OSCC) is one of the most common types of cancer worldwide. Accumulating evidence has shown that long noncoding RNAs (lncRNAs) serve important roles in the development of OSCC. The purpose of this study was to investigate the biological function and underlying regulatory mechanism of lncRNA homeobox A cluster antisense RNA2 (HOXA-AS2) in OSCC. RT-qPCR was performed to analyze the HOXA-AS2 expressions in human immortalized oral epithelial cell (HIOEC) line, human OSCC cell lines, and plasma. The expression of HOXA-AS2 and enhancer of zeste 2 polycomb repressive complex 2 subunit (EZH2) in Tca-8113 cells were knocked down or overexpressed by transfection with shRNA-HOXA-AS2 or pcDNA-EZH2, respectively. The interaction between HOXA-AS2 and EZH2 was validated by RNA immunoprecipitation assay. In addition, cell proliferation was assessed by CCK-8 and EdU assays. Cell cycle distribution was analyzed by flow cytometry. Cell migration and invasion were detected using wound healing and Transwell assays, respectively. Apoptosis was detected by TUNEL staining. The protein expression levels of cell cycle and apoptosis-related proteins were measured by western blot analysis. Compared with HIOEC cells, HOXA-AS2 expression in OSCC cells was upregulated. HOXA-AS2 knockdown significantly inhibited Tca-8113 cell proliferation, blocked the cell cycle by arresting cells in the G0/G1 phase, promoted apoptosis, and suppressed migration and invasion. In addition, HOXA-AS2 was predicted to directly target EZH2 and positively regulate EZH2 expression. EZH2 overexpression could reverse the inhibitory effect of HOXA-AS2 knockdown on the proliferation, migration, and invasion of Tca-8113 cells. In summary, the findings suggested that HOXA-AS2 may inhibit cell proliferation, invasion, and migration, induce cell cycle arrest in the G0/G1 phase, and increase cell apoptosis by targeting EZH2. The research indicated that HOXA-AS2/EZH2 axis may play a key role in the development of OSCC.

Knockdown of Long Non-Coding RNA (lncRNA) Colon Cancer-Associated Transcript-1 (CCAT1) Suppresses Oral Squamous Cell Carcinoma Proliferation, Invasion, and Migration by Inhibiting the Discoidin Domain Receptor 2 (DDR2)/ERK/AKT Axis.

BackgroundEmerging evidence shows that lncRNAs are involved in carcinogenesis or suppression in diverse cancers. This study assessed the biological role of lncRNA CCAT1 in OSCC and explored the underlying molecule mechanism.Material/MethodsCCAT1 and DDR2 expression was measured by qRT-PCR. Colony formation assay and CCK-8 assay were performed to evaluate cell proliferation. Cell cycle was determined by flow cytometric analysis and Western blot analysis. In addition, wound healing and Transwell assay were used to assess cell migration and invasion, respectively. RNA immunoprecipitation (RIP) assay were employed to identify the interaction between DDR2 and CCAT1. Protein levels involved in DDR2/ERK/AKT pathway were estimated by Western blot assay.ResultsThe findings revealed that CCAT1expression was upregulated in OSCC cell lines. Knockdown of CCAT1 repressed cell proliferation, blocked the cell cycle, and suppressed the invasion and migration of TCA-8113 cells. Moreover, DDR2 expression in OSCC cell lines was downregulated and CCAT1 silencing repressed the expression of DDR2. RIP assays validated the binding of CCAT1 and DDR2 protein. Moreover, CCAT1 silencing suppressed the ERK/AKT signaling through DDR2 in TCA-8113 cells.ConclusionsDownregulation of CCAT1 suppressed TCA-8113 cell proliferation, invasion, and migration by inactivation of the ERK/AKT pathway via inhibition of DDR2, suggesting the value of CCAT1 in diagnosis and treatment of patients with OSCC.MeSH Keywords: MAP Kinase Signaling System/Mouth Neoplasms/RNA, Long Noncoding

Matrine from Vietnamese sophora root inhibits the growth of oral squamous cell carcinoma cells in vitro and in vivo

Abstract This study investigated the extraction of matrine from Vietnamese sophora root and its in vitro and in vivo inhibitory effects on oral squamous cell carcinoma (OSCC) cells. Matrine was extracted from Vietnamese sophora root using microwave-assisted extraction technology and purified using macroporous adsorption resin. Human OSCC Tca8113 cells were treated with matrine with concentration of 0, 0.5, 1, 2, 4 and 8 g/L. The proliferation and apoptosis of cells were detected. OSCC cells were transplanted in the nude mice, and then the nude mice were treated with normal saline, cyclophosphamide (CTX) (20 mg·kg-1) and matrine (100 mg·kg -1), respectively. The optimized extraction parameters of matrine were obtained. The in vitro experiments showed that, matrine inhibit the proliferation of Tca8113 cells and induced the cell apoptosis. The in vivo experiments found that, the tumor inhibition rate of matrine was basically equivalent to that of CTX. In addition, compared with CTX, matrine had no obvious toxic side effect on the liver and kidney function of nude mice. These results indicate that, matrine has obvious in vitro and in vivo inhibitory effects on OSCC cells, with no obvious toxic side effect on the liver and kidney function.

Study of As2O3 regulating proliferation and apoptosis of Tca8113 cells by inhibiting the expression of Id-1

Objective: Our study aims to investigate the effect of arsenic trioxide (As2O3) on proliferation and apoptosis of Tca8113 tongue squamous carcinoma cells.Methods: Cell proliferation and the expression of Id-1 mRNA in Tca8113 cells after treatment with different concentrations of As2O3 were detected by MTT and qRT-PCR, respectively. The expression of Id-1, cell proliferation and apoptosis in Id-1 silencing Tca8113 cells were detected by qRT-PCR, Western blot, MTT and flow cytometry, respectively. The pcDNA 3.1-Id-1 overexpression vector was transfected into Tca8113 cells combination with 3 μmol/L As2O3. The detection of cell proliferation, apoptosis and Caspase-3, Bax and Bcl-2 protein expression in transfected Tca8113 cells were performed by MTT, flow cytometry and Western blot assay, respectively.Results: As2O3 of different concentration could inhibit the proliferation of Tca8113 cells and IC50 value was 3.004 ± 0.2379 μmol/L. The expression of Id-1 mRNA was down-regulated in Tca8113 cells treated with 3 μmol/L As2O3 for 48 h. The results of qRT-PCR, Western blot, MTT and flow cytometry indicated that the expression level of Id-1 and cell proliferation ability were decreased while the apoptosis rate was increased in Tca8113 cells after transfection of Id-1 siRNA. Overexpression of Id-1 could attenuate the inhibition or promotion of As2O3 on proliferation, apoptosis and Caspase-3, Bax and Bcl-2 protein expression in Tca8113 cells.Conclusion: As2O3 could regulate the proliferation and apoptosis of Tca8113 cells by inhibiting the expression of Id-1.

Effect of brazilin on apoptosis and autophagy of tongue cancer Tca8113 cells and its molecular mechanism

To investigate the effects of brazilin on the proliferation, apoptosis and autophagy of human tongue squamous cell carcinoma Tca8113 cells in vitro and explore its molecular mechanism. The changes in the proliferation, morphology and apoptosis of Tca8113 cells in response to brazilin treatment were detected using MTT assay, Hoechst33342 staining, and Annexin V/PI double staining, respectively. The expressions of apoptosis-related protein Bax, Bcl-2, cleaved caspase-3 and autophagy-related proteins p-AMPK, p-mTOR, LC3B, and p62 in the treated cells were detected using Western blotting. The effect of treatment with both the AMPK pathway inhibitor and brazilin on the expressions of the pathway-related proteins p-AMPK, p-mTOR, and LC3B was assessed. MTT assay showed that brazilin significantly inhibited the proliferation of Tca8113 cells with an IC50 of 31.17 μmol/L at 24 h. Hoechst33342 staining showed that brazilin induced apoptotic morphological changes in Tca8113 cells in a concentration-dependent manner. Treatment with different concentrations of brazilin resulted in increased apoptosis in the cells. Brazilin obviously inhibited the expression of Bcl-2, p62 and p-mTOR and enhanced the expressions of Bax, cleaved caspase-3, LC3B and p-AMPK. The AMPK pathway inhibitor significantly inhibited the increase in p-AMPK and LC3B expressions and the decrease in p-mTOR expression induced by brazilin. Brazilin can inhibit the proliferation and promote apoptosis in Tca8113 cells and at the same time induces autophagy in the cells through the AMPK/mTOR pathway.

CXCL14 regulates cell proliferation, invasion, migration and epithelial-mesenchymal transition of oral squamous cell carcinoma

Oral squamous cell carcinoma (OSCC) is an aggressive malignancy which is the most common type of head and neck cancer. The study aimed to investigate the role of CXCL14 in cell proliferation, invasion, migration, and epithelial-mesenchymal transition (EMT) of OSCC and its potential mechanisms. In the present study, overexpression of CXCL14 was employed in TCA-8113 cells. Cell proliferation was detected using Cell Counting Kit-8 assay and flow cytometry. Scratch wound healing and Transwell assays were applied for the analysis of cell migration and invasion, respectively. Quantitative real-time polymerase chain reaction analysis was used to explore the mRNA expression of CXCL14 and PD-L1. Moreover, the protein expression levels of CXCL14, E-cadherin, N-cadherin, vimentin, PD-L1, NF-κB p65, and phospho-IκB-α (p-IκB-α) were examined by Western blot. The results revealed that the overexpression of CXCL14 was associated with suppressed proliferation of TCA-8113 cells coupled with decreasing the percentages of cells in the S phase. Moreover, the levels of cell migration and invasion were attenuated as well. Additionally, the expression level of EMT relative protein E-cad was increased notably, whereas N-cad and Vimentin were decreased. Importantly, the mRNA and protein expression of PD-L1, a crucial factor for OSCC, were lowered after overexpression of CXCL14. At the same time, the expression of NF-κB p65 and p-IκB-α was down-regulated significantly. Taken together, we speculate that CXCL14 inhibited OSCC cell proliferation, invasion, migration and EMT via suppressing PD-L1 expression and NF-κB mediated EMT, which indicated that CXCL14 may be an underlying target for OSCC treatment.

HIF-1α restricts proliferation and apoptosis of Tca8113 cells through up regulation of Hippo signaling pathway under hypoxic conditions.

The hypoxia-inducible factor-1α (HIF-1α) is a key factor for tumor cells adaptation to hypoxia. Studies have shown that under hypoxic conditions, HIF-1α expression was significantly increased in human tongue squamous cell carcinoma cells (Tca8113). This research aims to determine the exact mechanism of HIF-1α on the proliferation and apoptosis of Tca8113 cells. Tca8113 cells were cultured under normoxia and hypoxia. Real Time-PCR and Western blot were used to measure the expression levels of HIF-1α and TAZ. Under hypoxic condition, HIF-1α siRNA was transfected into Tca8113 cells. CCK8 was used to measure the proliferation of Tca8113 cells. Flow cytometry was used to detect apoptosis of Tca8113 cells. Under hypoxic condition, the expression levels of HIF-1α and TAZ at both mRNA and protein levels were significantly increased (p <0.05). The downregulation of HIF-1α by siRNA significantly inhibited Tca8113 cells proliferation, increased their apoptosis, and reduced the expression level of TAZ. Under hypoxic conditions, HIF-1α inhibits the proliferation and apoptosis of Tca8113 cells via the elevation of the Hippo signaling pathway.

Knockdown of IFI27 inhibits cell proliferation and invasion in oral squamous cell carcinoma

BackgroundThe development of oral squamous cell carcinoma (OSCC) involves genetic mutations, epigenetic gene expression modification, and other processes. It has been reported that IFI27 is upregulated in OSCC, but its function is unknown. The aim of this study was to investigate the role of IFI27 on OSCC cell proliferation and invasion.MethodsThe protein level of IFI27 in OSCC tissues and adjacent tissues was detected by immunohistochemistry. In the OSCC cell model, we designed the IFI27 siRNA to downregulate the expression of IFI27; gene and protein of IFI27 in those models were then detected by Q-PCR and Western blot. MTT assay was used to detect the effect of -IFI27 knockdown on cell proliferation; Annexin V-PI staining flow cytometry was used to detect the effect of IFI27 downregulation on apoptosis of cancer cells. The effect of IFI27 downregulation on oral cancer cell invasion was detected using Transwell assay.ResultsIFI27 was highly expressed in OSCC tissues by immunohistochemical assay. In the OSCC cell model, IFI27 siRNA could downregulate the mRNA and protein expression level of IFI27. As showed in MTT assay, Annexin V-PI assay, and Transwell assay, through the downregulation of IFI27, TSCCA and TCA8113 cell proliferation were inhibited, OSCC cell apoptosis was promoted, and its migration and invasion were inhibited.ConclusionIFI27 is involved in the development and progression of OSCC. Its high expression promotes cell proliferation and invasion and reduces apoptosis. These findings may provide new biomarkers and therapeutic targets for OSCC diagnosis and clinical treatment.

Association between integrin-linked kinase and hyperthermia in oral squamous cell carcinoma.

The present study aimed to observe the effect of the biological functions of integrin-linked kinase (ILK) silencing combined with hyperthermia on Tca8113 cells. Lentivirus-mediated short hairpin RNA (shRNA)-targeting ILK was transfected into oral squamous cell carcinoma (OSCC) Tca8113 cells and, combined with hyperthermia, several experimental methods were used to detect their biological behavior in vitro. On the basis of in vitro experiments, Tca8113 cells were transplanted into nude mice models, and ILK-shRNA-lentivirus was injected into the nude mice transplanted tumor and combined with hyperthermia. Tumor morphology and the associated protein expression changes were determined. Subsequent to ILK silencing combined with hyperthermia, the growth, migration and proliferation of Tca8113 cells were significantly inhibited. Flow cytometry revealed that the cells were blocked in the S phase, and western blot analysis demonstrated that ILK, phosphorylated (p)-RAC-alpha serine/threonine-protein kinase (Akt), p-glycogen synthase kinase-3β and p-heat shock factor 1 protein expression levels were significantly decreased, while apoptosis-associated protein B-cell lymphoma-2-associated X protein expression and the efficacy of hypothermia were significantly increased. By ILK silencing combined with hyperthermia, a significant therapeutic effect on transplanted tumors was observed in nude mice. Immunohistochemistry revealed the same results as the in vitro experiments. ILK silencing combined with hyperthermia can inhibit the growth, proliferation and migration of Tca8113 cells, promote Tca8113 cell apoptosis, inhibit the phosphatidylinositol-3-kinase/Akt signaling pathway and increase hyperthermia sensitivity; the combination therapy exhibits a synergistic sensitizing effect. Therefore, ILK silencing combined with hypothermia may serve as a novel combination therapy strategy against OSCC.

The EP4 antagonist, L-161,982, induces apoptosis, cell cycle arrest, and inhibits prostaglandin E2-induced proliferation in oral squamous carcinoma Tca8113 cells.

Recent studies suggest that cyclooxygenase 2 (COX-2) inhibitors may enhance the toxic effects of anticancer drugs on tumor cells, including oral squamous cell carcinoma (OSCC), but its long-term use can cause side effects such as stomach ulcers and myocardial infarction. Our aim was to investigate proliferative effects of a downstream product of COX-2, prostaglandin E2 (PGE2), in human oral squamouscarcinoma cell line Tca8113 and explore the effects of PGE2 receptors, especially EP4 receptor, on the growth of Tca8113 cells. To evaluate the effects of PGE2 and EP receptors on Tca8113 cells, CCK8 assay, Western blotting, cell cycle analysis, and apoptosis assay were performed. We found that the EP4 receptor agonist, PGE1-OH, could mimick PGE2 rescued the inhibitory effect of celecoxib and induced cell growth via ERK phosphorylation, and the EP4 receptor antagonist, L-161,982, completely blocked PGE2-stimulated ERK phosphorylation and proliferation of Tca8113 cells. Furthermore, L-161,982 may induce apoptosis and block cell cycle progression at s phase by upregulating Bax and p21 protein levels and by downregulating Bcl-2, CDK2, and cyclin A2 protein levels. Our results indicate that EP4 receptor mediates PGE2-induced cell proliferation through ERK signaling, and inhibition of EP4 receptor may represent an alternative therapeutic strategy for the prevention and treatment of OSCC.

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.

Synthetic Isoliquiritigenin Inhibits Human Tongue Squamous Carcinoma Cells through Its Antioxidant Mechanism.

Isoliquiritigenin (ISL), a natural antioxidant, has antitumor activity in different types of cancer cells. However the antitumor effect of ISL on human tongue squamous carcinoma cells (TSCC) is not clear. Here we aimed to investigate the effects of synthetic isoliquiritigenin (S-ISL) on TSCC and elucidate the underlying mechanisms. S-ISL was synthesized and elucidated from its nuclear magnetic resonance spectrum and examined using high performance liquid chromatography. The effects of S-ISL on TSCC cells (Tca8113) were evaluated in relation to cell proliferation, apoptosis and adhesion, migration, and invasion using sulforhodamine B assay, fluorescence microscopy technique, flow cytometry (FCM) analysis, and Boyden chamber assay. The associated regulatory mechanisms were examined using FCM and fluorescence microscopy for intracellular reactive oxygen species (ROS) generation, Gelatin zymography assay for matrix metalloproteinase (MMP) activities, and Western blot for apoptosis regulatory proteins (Bcl-2 and Bax). Our data indicated that S-ISL inhibited Tca8113 cell proliferation, adhesion, migration, and invasion while promoting the cell apoptosis. Such effects were accompanied by downregulation of Bcl-2 and upregulation of Bax, reduction of MMP-2 and MMP-9 activities, and decreased ROS production. We conclude that S-ISL is a promising agent targeting TSCC through multiple anticancer effects, regulated by its antioxidant mechanism.

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

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. 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 β-catenin and TGF-β1 were analyzed by Western blot. 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 β-catenin were reduced, whereas the expression levels of vimentin and TGFβ-1 were elevated in the PTEN-siRNA group compared with those of control and negative groups. These results were significantly different. 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.

HMME combined with green light-emitting diode irradiation results in efficient apoptosis on human tongue squamous cell carcinoma.

Hematoporphyrin monomethyl ether (HMME) is a novel and promising porphyrin-related photosensitizer for photodynamic therapy (PDT). This study aimed to investigate the efficacy and potential mechanism of HMME-PDT under irradiation of green light-emitting diode (LED) with wavelength of 530 ± 20nm in treating human tongue squamous cell carcinoma Tca8113 cells in vitro. The HMME concentrations were 1.25, 2.5, and 5μg/ml while the energy densities were 0.6, 1.2, 1.8, 2.4, and 3.0J/cm(2). MTT assay demonstrated that HMME-PDT significantly inhibited the proliferation of Tca8113 cells, and the cytotoxicity was improved with increased HMME concentration and light intensity. The amount of cells decreased significantly and the morphology of cells changed drastically after HMME-PDT. Flow cytometry analysis revealed that HMME-PDT induced both apoptosis and necrosis, but apoptosis was the main form of cell death. Apoptotic morphology was confirmed by Hoechst 33342 staining. Laser scanning confocal microscopy observation showed that HMME was mainly localized in mitochondria. The production of intracellular reactive oxygen species increased remarkably after PDT treatment, and both sodium azide (the singlet oxygen quencher) and D-mannitol (the hydroxyl radical scavenger) could protect Tca8113 cells from death induced by HMME-PDT. Additionally, the activity of caspase-3 also increased markedly in treated groups, and the cell death could be rescued by a reversible inhibitor (Ac-DEVD-CHO) of caspase-3. These results demonstrated that HMME combined with green LED significantly induced apoptosis of Tca8113 cells, suggesting that HMME-PDT using green LED might be a potential therapeutic strategy for human tongue squamous cell carcinoma.

Down-Regulating Receptor Interacting Protein Kinase 1 (RIP1) Promotes Oxaliplatin-Induced Tca8113 Cell Apoptosis.

BackgroundOxaliplatin is a crucial chemotherapy drug that plays an important role in colorectal cancer and oral cancer treatment. However, the molecular mechanism of oxaliplatin in killing tongue squamous cell cancer cells is still unknown. This paper investigates the mechanism of by which oxaliplatin regulates tongue squamous cell carcinoma Tca8113 cell survival and death.Material/MethodsTca8113 was treated with 1 μmol/L oxaliplatin for 24 h. Tca8113 cell proliferation and apoptosis were determined by MTT method and flow cytometry, respectively. Western blot was applied to detect receptor-interacting protein kinase 1 (RIP1) level. Tca8113 was transfected with siRNA RIP1 and then treated with 1 μmol/L oxaliplatin, and the cell apoptosis was detected.ResultsWe found that 1 μmol/L oxaliplatin could inhibit Tca8113 cell growth (cell survival rate was 19.3%), reduce mitochondrial membrane potential (reduce 82.3%) and phosphatidylserine eversion (positive rate was 62.7%), and activate caspase-3 (increased 2.6 times). We also found that 1 μmol/L oxaliplatin treatment could increase RIP1 expression in Tca8113 cells. Cell apoptosis rate increased after siRNA RIP1 and 1 μmol/L oxaliplatin treatment (apoptosis rate was 90.2%).ConclusionsDown-regulating RIP1 promotes oxaliplatin induced Tca8113 cells apoptosis.

AS-miR-222 transfection target puma to induce cell apoptosis in oral squamous cell carcinoma

Objective To investigate the effect of overexpression of p53 upregulated modulator of apoptosis (PUMA) gene induced by As-miR-222 transfection on apoptosis of oral squamous cell carcinoma(OSCC). Methods Tca8113 was treated with As-miR-222 and As-miR-222 NC respectively , the experiment was randomly divided into 3 groups : blank group, As-miR-222 transfection group and As-miR-222 NC transfection group. After the recombinants were transfected into Tca8113 with Lipofectamine TM MAX, the expression of PUMA on mRNA and protein level was analyzed by RT-PCR and Western blot, respectively. The proliferation of Tca8113 cells was detected by MTT assay. Cell cycle and apoptosis were detected by flow cytometry. Results Compared with blank group and As-miR-222 NC transfection group, miR-222 expression was down-regulated obviously (P 0.01) meanwhile the expression of PUMA at both mRNA level and protein level was up-regulated as validated by RT-PCR and Western blot (P 0.05). MTT assay indicated that cell proliferation was inhibited by As-miR-222 transfection. Cell apoptosis ability was significantly increased after treatment with As-miR-222 . Conclusion As-miR-222 transfection could up-regulate the expression of PUMA to induce apoptosis in human oral squamous cell carcinoma.

Stable gene-silence of Kif2a synergistic with 5-fluorouracil suppresses oral tongue squamous cell carcinoma growth in vitro and in vivo

Squamous cell carcinoma of the oral tongue (SCCOT) is one of the most common malignant carcinomas in the head and neck. Recurrence and/or metastasis often results in failure of treatment and decreases the survival of the patients. The purpose of this study is to investigate the effect of gene-silence of Kif2a on SCCOT in viro and in vivo. Plasmid-mediated expression of Kif2a-siRNA (pGPU6/GFP/Kif2a) was employed to silence the expression of Kif2a in Tca8113 cells at both mRNA and protein levels. Tca8113 cell proliferation was measured by 3-(4,5-dimethylthiazol-2-yl)-2, 5-diphenyltetrazolium bromide (MTT) assay and growth of Tca8113 tumors was determined by intra-tumor injection of pGPU6/GFP/Kif2a in nude mice. Gene-silence of Kif2a suppressed Tca8113 cell proliferation. pGPU6/GFP/Kif2a synergized the tumor suppression effect of 5-Fluorouracil (5-Fu) on Tca8113 cells. Our data support that Kif2a is a potential molecular target for the therapeutics of recurrent and metastatic SCCOT.

Knockdown of Cdc6 Inhibits Proliferation of Tongue Squamous Cell Carcinoma Tca8113 Cells

The present study aimed at evaluating the effects of Cdc6 downregulation on the proliferation of Tca8113 cells. Two lentiviral vectors (KD1 and KD2) expression cdc6 siRNA were constructed and then infected into Tca8113 cells. Real-time PCR and Western blot analysis were performed to detect the mRNA and protein expression of Cdc6. MTT assays were employed to delineate the growth curves, and flow cytometry was performed to assess cell-cycle progression and apoptosis in Tca8113 cells. Following infection with the lentiviral vectors, real-time PCR and Western blot analysis revealed that Cdc6 expression was markedly suppressed in Tca8113 cells. When compared with the negative control group, the mRNA expression of Cdc6 was reduced by 50% and 65% and the protein expression by 65.87% and 79.38% in cells harboring KD1 or KD2, respectively. Cell growth was slowed, and the growth inhibition rate was 25.84% and 30.34% in Tca8113 cells following infection with KD1 or KD2, respectively. In addition, cell-cycle progression was altered. In KD- infected Tca8113 cells, the proportion of cells in the S phase was markedly reduced, but the proportion in the G1 phase was significantly increased; this was accompanied by an increase in cell apoptosis. Downregulation of Cdc6 effectively inhibited the proliferation of Tca8113 cells.