Squamous Carcinoma Cell Line Tca8113 Research Articles

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.

Upregulation of PTEN suppresses invasion in Tca8113 tongue cancer cells through repression of epithelial-mesenchymal transition (EMT).

We previously discovered that the expression of the tumor suppressor phosphatase and tensin homolog (PTEN) was downregulated in the majority patients with tongue squamous cell carcinoma (TSCC). The aim of this study was to investigate the role of PTEN overexpression in the regulation of epithelial-mesenchymal transition (EMT) of the tongue squamous carcinoma cell line Tca8113 as well as explore the underlying mechanism. GV230 (containing the PTEN gene) and empty vectors were transfected into Tca8113 cells. After stable transfection, the messenger RNA (mRNA) and protein levels of PTEN were validated using quantitative real-time PCR (qPCR) and Western blot analysis. The growth and cell cycle were analyzed using Cell Counting Kit-8 (CCK-8) and flow cytometry, respectively. The invasion ability was measured with a transwell assay. The effects of PTEN overexpression on EMT and Hedgehog signaling were assessed by comparing Tca8113-PTEN cells with control and negative control cell groups. We found that PTEN expression was significantly upregulated after transfection. Meanwhile, upregulated PTEN inhibited the proliferation and invasion of Tca8113 cells. In addition, we observed changes in the EMT- and Hedgehog-associated proteins. These data demonstrated that PTEN upregulation could reduce invasion by inhibiting the process of EMT in Tca8113 cells, which might be related to the Hedgehog signaling pathway.

Effects of sodium butyrate on proliferation of human oral squamous carcinoma cell line and expression of p27Kip1

To investigate the effect of sodium butyrate (NaB) on inhibition of human oral squamous carcinoma cell line. Human oral squamous carcinoma cell line Tca8113 was treated with different concentration of NaB. Light microscope was used to observe the morphological changes of the carcinoma cells. The cell cycle was analyzed by flow cytometry. Expression of p27(Kip1) was determined with immunocytochemical assay. NaB significantly inhibited the proliferation of Tca8113 in a time- and dose-dependent manner. Tca8113 cells treated with NaB was arrested in G(0)-G(1) phase. The fraction of cells in G(0)-G(1) were (63.2 ± 2.4)% and (77.2 ± 3.8)% after treated with NaB at the concentration of 2 and 4 mmol/L alternatively, whereas (48.1 ± 2.4)% in G(0)-G(1) phase were observed in control group (P < 0.05). The expression of p27(Kip1) was markedly up-regulated after being treated with NaB. NaB treatment can inhibit the growth of oral squamous carcinoma cell line in vitro and induce cell cycle arrest, which might be associated with the increased expression of p27(Kip1) protein.

Inhibitive effect of celecoxib on the adhesion and invasion of human tongue squamous carcinoma cells to extracellular matrix via down regulation of MMP-2 expression

The effect of cyclooxygenase 2 (COX-2) and its inhibitor on metastasis and invasion of tumor and its relationship with matrix metalloproteinase are getting more and more attention. Down regulation of matrix metalloproteinase (MMP) expression could inhibit the invasion and adhesion ability of cancer cells. Our study observed the inhibitive effect of selective COX-2 inhibitor, celecoxib, on adhesion and invasion ability of human tongue squamous carcinoma cell line Tca8113 and its relationship with MMP-2 secretion. COX-2 expression of Tca8113 cell was detected by Western blot analysis. Ratio of COX-2 positive Tca8113 cell was determined by flow cytometry analysis. MMP-2 and PGE2 secretion was measured by ELISA. Invasion of a Matrigel-coated membrane by Tca8113 cells was quantitatively assessed by counting migrated cells. Results showed that COX-2 expression was positive in Tca8113 cell and could be significantly inhibited by celecoxib. Celecoxib could inhibit PGE2 production of Tca8113 cell in a dose-dependent manner, down-regulate MMP-2 secretion of Tca8113 cell, and at the same time significantly inhibit invasion and adhesion ability of these cells. Adding extrinsic PGE2 could antagonize the inhibitive effect of celecoxib not only on MMP-2 secretion of Tca8113 cell, but also on invasion and adhesion ability of the cells. The results suggested that celecoxib had chemoprevention effect on invasion and metastasis of oral carcinoma.

Celecoxib enhances the lethal effects of bleomycin in human tongue squamous carcinoma cell line Tca8113

To investigate the role of COX-2 inhibitor celecoxib in enhancing the lethal effects of bleomycin in Tca8113 cell line. Tca8113 cells were treated with different concentrations of celecoxib and bleomycin for 24, 48, 72 h. Methyl thiazolyl tetrazolium assay was used to calculate cell growth inhibition rate and Jin Zheng Jun's method was used to evaluate the interaction of celecoxib and bleomycin on Tca8113 cells. Flow cytometry was used to evaluate the effects of combined use of celecoxib and bleomycin on cell cycle progress and apoptosis. Low dose of celecoxib (10 micromol/L, < IC(50)) combined with bleomycin showed synergism or additive lethal effect on Tca8113 cell line. Celecoxib could notably enhance the inhibitory effect of bleomycin on Tca8113 cells by blocking cell cycle progress and thus resulting in the increasing G(0)/G(1) cells [(60.93 +/- 0.32)%] distribution and inducing apoptosis [(1.87 +/- 0.11)%]. Low doses of celecoxib could significantly enhance the lethal effect of bleomycin on Tca8113 cells by inhibiting cell growth and proliferation through blocking cell cycle progress and inducing apoptosis. The ways of these interactions on inhibiting Tca8113 cell growth were synergistic or/and additive.

Effect of using RNA interference to alter iNOS gene expression on the proliferation of tongue squamous cell carcinoma cell line Tca8113

This study used RNA interference (RNAi) to explore the effect of NO and inducible nitric oxide synthase (iNOS) on apoptosis and proliferation in the tongue squamous carcinoma cell line Tca8113. Tca8113 cells were transfected with the plasmid pGenesil-1, which expresses iNOS short hairpin RNA (shRNA), or the negative control plasmid pSilencer-HK, and the transfected cells were compared with untransfected cells. The expression of iNOS was detected by histochemistry, and apoptosis was detected by flow cytometry. The expression of iNOS was significantly lower in the pSilencer-iNOS group than in the pSilencer-HK and empty control groups. The apoptosis rate was significantly higher in the pSilencer-iNOS group than in the pSilencer-HK and empty control groups. Growth monitoring showed that proliferation was also inhibited in cells transfected with pSilencer-iNOS. RNAi gene silencing decreased iNOS gene expression, induced apoptosis, and suppressed proliferation in Tca8113 cells.

Effects of rAAV-CD151 and rAAV-antiCD151 on the migration of human tongue squamous carcinoma cell line Tca8113.

This study was designed to determine the effects of the recombinant adeno-associated virus vector containing sense CD151 gene (rAAV-CD151) and antisense CD151 gene (rAAV-antiCD151) on the migration of Tca8113 cell. Functional fragment of CD151 gene was amplified by RT-PCR, and inserted into the vector pAAV in the sense direction and antisense direction, respectively. The rAAV-CD151 and rAAV-antiCD151 were produced and the titers were determined by dot blot. The CD151, at protein level, was detected by Western blot. The Transwell chamber was used to detect the effects of the rAAV-CD151 and rAAV-antiCD151 on the tumor cell migration. The titers of the rAAV-CD151 and rAAV-antiCD151 were 2 x 10(11) pfu/ml and 1.0 x 10(11) pfu/ml, respectively. The expression of CD151 was increased by 108% in the cells transfected with rAAV-CD151 and decreased by 79% in the cells transfected with rAAV-antiCD151, as compared with non-transfected cells, respectively. The number of the migrating cells was significantly increased in the cells transfected with rAAV-CD151 (93.56 +/- 11.59) and decreased in the cells transfected with rAAV-antiCD151 (24.00 +/- 4.36) as compared with non-transfected and rAAV-GFP transfected cells (53.00 +/- 6.56 and 46.00 +/- 7.00, P<0.05). It is an important molecular mechanism of the tumor metastasis that the overexpression of CD151 promotes the migration of the tumor cells. The rAAV-antiCD151 is a novel tool, which can reduce the expression of CD151 and inhibit the migration of the tumor cells, and brings us a new approach of anti-sene gene therapy targeted at CD151 in human carcinoma.