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Apoptotic Effect of Tolfenamic Acid in KB Human Oral Cancer Cells: Possible Involvement of the p38 MAPK Pathway.

Kim JH, Jung JY, Shim JH, Kim J, Choi KH, Shin JA, Choi ES, Lee SO, Chintharlapalli S, Kwon KH, Leem DH, Cho NP, Cho SD - J Clin Biochem Nutr (2010)

Bottom Line: The results showed that tolfenamic acid does not alter the expression of the COX proteins, but it inhibits cell growth and induces apoptosis as evidenced by the annexin V positivity, sub-G(1) population, nuclear fragmentation and the cleavage of poly ADP-ribose polymerase.In addition, tolfenamic acid also leads to a loss of the mitochondrial membrane potential in KB cells.These effects are related to the activation of p38 mitogen-activated protein kinase (MAPK) pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral Pathology, School of Dentistry and Institute of Oral Bioscience, Brain Korea 21 project, Chonbuk National University, Jeonju 561-756, Republic of Korea.

ABSTRACT
Nonsteroidal anti-inflammatory drugs (NSAIDs) are known to inhibit cancer growth by inhibiting the activity of cyclooxygenase (COX). However, there is increasing evidence that the COX-independent pathway may be also involved in the inhibitory effect of NSAIDs against tumor progression. Tolfenamic acid is a NSAID that exhibits anticancer activity in pancreatic and colorectal cancer models. In the present study, the anti-tumor effect of tolfenamic acid in KB human oral cancer cells is investigated. The results showed that tolfenamic acid does not alter the expression of the COX proteins, but it inhibits cell growth and induces apoptosis as evidenced by the annexin V positivity, sub-G(1) population, nuclear fragmentation and the cleavage of poly ADP-ribose polymerase. In addition, tolfenamic acid also leads to a loss of the mitochondrial membrane potential in KB cells. These effects are related to the activation of p38 mitogen-activated protein kinase (MAPK) pathway. These results suggest that tolfenamic acid-induced apoptotic cell death inhibits cancer growth by activating the p38 MAPK pathway for cancer prevention.

No MeSH data available.


Related in: MedlinePlus

The effect of tolfenamic acid on apoptotic cell death in KB cells. The KB cells were treated with dimethyl sulfoxide (DMSO) or various concentrations of 75 and 100 µM of tolfenamic acid for 48 h. A, The sub-G1 fraction was assessed by propidium iodide (PI) staining and flow cytometry analysis, Bar graph represents two-independent experiments; B, Annexin V-PI staining was carried out to detect apoptosis, Bar graph represents two-independent experiments; C, Fluorescence microscopy images of the 4'-6-diamidino-2-phenylindole (DAPI)-stained KB cells showing the concentration-dependent appearance of an apoptotic morphology in the tolfenamic acid-treated KB cells. White arrows point nuclear condensation and fragmentation.
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Figure 3: The effect of tolfenamic acid on apoptotic cell death in KB cells. The KB cells were treated with dimethyl sulfoxide (DMSO) or various concentrations of 75 and 100 µM of tolfenamic acid for 48 h. A, The sub-G1 fraction was assessed by propidium iodide (PI) staining and flow cytometry analysis, Bar graph represents two-independent experiments; B, Annexin V-PI staining was carried out to detect apoptosis, Bar graph represents two-independent experiments; C, Fluorescence microscopy images of the 4'-6-diamidino-2-phenylindole (DAPI)-stained KB cells showing the concentration-dependent appearance of an apoptotic morphology in the tolfenamic acid-treated KB cells. White arrows point nuclear condensation and fragmentation.

Mentions: To determine if tolfenamic acid induced apoptosis of the KB cells, the level of poly ADP-ribose polymerase (PARP) cleavage was examined by Western blot anlaysis (Fig. 2D). After the tolfenamic acid treatment for 48 h, the cleaved PARP (85 kDa) was clearly detected in the cells exposed to 75 and 100 µM of tolfenamic acid whereas the cleaved PARP was not observed in DMSO-treated KB cells. To confirm the apoptotic activity of tolfenamic acid, other distinct features of apoptosis (e.g. Annexin V positivity, sub-G1 population and nuclear fragmentation using DAPI staining) were examined (Fig. 3). The sub-G1 peak was not detected in DMSO-treated KB cells. However, 48 h of exposure to 75 and 100 µM of tolfenamic acid in KB cells resulted in cell accumulation in the sub-G1 phase in a concentration-dependent manner. In addition, 75 and 100 µM of tolfenamic acid increased the number of annexin V-positive KB cells and nuclear fragmented cells when compared to the DMSO-treated KB cells. This suggests that apoptotic cell death event contributed to the growth-inhibitory effect of tolfenamic acid in KB cells.


Apoptotic Effect of Tolfenamic Acid in KB Human Oral Cancer Cells: Possible Involvement of the p38 MAPK Pathway.

Kim JH, Jung JY, Shim JH, Kim J, Choi KH, Shin JA, Choi ES, Lee SO, Chintharlapalli S, Kwon KH, Leem DH, Cho NP, Cho SD - J Clin Biochem Nutr (2010)

The effect of tolfenamic acid on apoptotic cell death in KB cells. The KB cells were treated with dimethyl sulfoxide (DMSO) or various concentrations of 75 and 100 µM of tolfenamic acid for 48 h. A, The sub-G1 fraction was assessed by propidium iodide (PI) staining and flow cytometry analysis, Bar graph represents two-independent experiments; B, Annexin V-PI staining was carried out to detect apoptosis, Bar graph represents two-independent experiments; C, Fluorescence microscopy images of the 4'-6-diamidino-2-phenylindole (DAPI)-stained KB cells showing the concentration-dependent appearance of an apoptotic morphology in the tolfenamic acid-treated KB cells. White arrows point nuclear condensation and fragmentation.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC2901767&req=5

Figure 3: The effect of tolfenamic acid on apoptotic cell death in KB cells. The KB cells were treated with dimethyl sulfoxide (DMSO) or various concentrations of 75 and 100 µM of tolfenamic acid for 48 h. A, The sub-G1 fraction was assessed by propidium iodide (PI) staining and flow cytometry analysis, Bar graph represents two-independent experiments; B, Annexin V-PI staining was carried out to detect apoptosis, Bar graph represents two-independent experiments; C, Fluorescence microscopy images of the 4'-6-diamidino-2-phenylindole (DAPI)-stained KB cells showing the concentration-dependent appearance of an apoptotic morphology in the tolfenamic acid-treated KB cells. White arrows point nuclear condensation and fragmentation.
Mentions: To determine if tolfenamic acid induced apoptosis of the KB cells, the level of poly ADP-ribose polymerase (PARP) cleavage was examined by Western blot anlaysis (Fig. 2D). After the tolfenamic acid treatment for 48 h, the cleaved PARP (85 kDa) was clearly detected in the cells exposed to 75 and 100 µM of tolfenamic acid whereas the cleaved PARP was not observed in DMSO-treated KB cells. To confirm the apoptotic activity of tolfenamic acid, other distinct features of apoptosis (e.g. Annexin V positivity, sub-G1 population and nuclear fragmentation using DAPI staining) were examined (Fig. 3). The sub-G1 peak was not detected in DMSO-treated KB cells. However, 48 h of exposure to 75 and 100 µM of tolfenamic acid in KB cells resulted in cell accumulation in the sub-G1 phase in a concentration-dependent manner. In addition, 75 and 100 µM of tolfenamic acid increased the number of annexin V-positive KB cells and nuclear fragmented cells when compared to the DMSO-treated KB cells. This suggests that apoptotic cell death event contributed to the growth-inhibitory effect of tolfenamic acid in KB cells.

Bottom Line: The results showed that tolfenamic acid does not alter the expression of the COX proteins, but it inhibits cell growth and induces apoptosis as evidenced by the annexin V positivity, sub-G(1) population, nuclear fragmentation and the cleavage of poly ADP-ribose polymerase.In addition, tolfenamic acid also leads to a loss of the mitochondrial membrane potential in KB cells.These effects are related to the activation of p38 mitogen-activated protein kinase (MAPK) pathway.

View Article: PubMed Central - PubMed

Affiliation: Department of Oral Pathology, School of Dentistry and Institute of Oral Bioscience, Brain Korea 21 project, Chonbuk National University, Jeonju 561-756, Republic of Korea.

ABSTRACT
Nonsteroidal anti-inflammatory drugs (NSAIDs) are known to inhibit cancer growth by inhibiting the activity of cyclooxygenase (COX). However, there is increasing evidence that the COX-independent pathway may be also involved in the inhibitory effect of NSAIDs against tumor progression. Tolfenamic acid is a NSAID that exhibits anticancer activity in pancreatic and colorectal cancer models. In the present study, the anti-tumor effect of tolfenamic acid in KB human oral cancer cells is investigated. The results showed that tolfenamic acid does not alter the expression of the COX proteins, but it inhibits cell growth and induces apoptosis as evidenced by the annexin V positivity, sub-G(1) population, nuclear fragmentation and the cleavage of poly ADP-ribose polymerase. In addition, tolfenamic acid also leads to a loss of the mitochondrial membrane potential in KB cells. These effects are related to the activation of p38 mitogen-activated protein kinase (MAPK) pathway. These results suggest that tolfenamic acid-induced apoptotic cell death inhibits cancer growth by activating the p38 MAPK pathway for cancer prevention.

No MeSH data available.


Related in: MedlinePlus