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Para-toluenesulfonamide induces tongue squamous cell carcinoma cell death through disturbing lysosomal stability.

Liu Z, Liang C, Zhang Z, Pan J, Xia H, Zhong N, Li L - Anticancer Drugs (2015)

Bottom Line: Para-toluenesulfonamide (PTS) has been implicated with anticancer effects against a variety of tumors.Next, the effects of PTS on cell viability, invasion, and cell death were determined.Lysosomal integrity assay and western blot showed that PTS increased lysosomal membrane permeabilization associated with activation of lysosomal cathepsin B.

View Article: PubMed Central - PubMed

Affiliation: aDepartment of Head and Neck Oncology bState Key Laboratory of Oral Diseases cDepartment of Oral and Maxillofacial Surgery, West China Hospital of Stomatology, Sichuan University, Chengdu dState Key Laboratory of Respiratory Disease, Guangzhou Institute of Respiratory Diseases, First Affiliated Hospital, Guangzhou Medical University, Guangzhou, China.

ABSTRACT
Para-toluenesulfonamide (PTS) has been implicated with anticancer effects against a variety of tumors. In the present study, we investigated the inhibitory effects of PTS on tongue squamous cell carcinoma (Tca-8113) and explored the lysosomal and mitochondrial changes after PTS treatment in vitro. High-performance liquid chromatography showed that PTS selectively accumulated in Tca-8113 cells with a relatively low concentration in normal fibroblasts. Next, the effects of PTS on cell viability, invasion, and cell death were determined. PTS significantly inhibited Tca-8113 cells' viability and invasive ability with increased cancer cell death. Flow cytometric analysis and the lactate dehydrogenase release assay showed that PTS induced cancer cell death by activating apoptosis and necrosis simultaneously. Morphological changes, such as cellular shrinkage, nuclear condensation as well as formation of apoptotic body and secondary lysosomes, were observed, indicating that PTS might induce cell death through disturbing lysosomal stability. Lysosomal integrity assay and western blot showed that PTS increased lysosomal membrane permeabilization associated with activation of lysosomal cathepsin B. Finally, PTS was shown to inhibit ATP biosynthesis and induce the release of mitochondrial cytochrome c. Therefore, our findings provide a novel insight into the use of PTS in cancer therapy.

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Para-toluenesulfonamide (PTS) selectively accumulates in Tca-8113 cells. (a) Tca-8113 were treated with PTS or dimethyl sulfoxide (control vehicle) and then subjected to high-performance liquid chromatography. Peak 1: antipyrine (internal standard), peak 2: PTS. (b) Plot of intracellular PTS concentration after 2.5–80 μmol/l PTS treatment for different periods of time in Tca-8113 cells. (c) Plot of intracellular PTS concentration after 2.5–80 μmol/l PTS treatment for different periods in human gingival fibroblast (HGF) cells. (d) Comparative evaluation of the intracellular PTS concentration under the same treatment condition in Tca-8113 and HGF cells. The 10 min time-point was chosen to observe the difference in the intracellular PTS concentration between Tca-8113 and HGF cells. Results are representative of three independent experiments. *P<0.05.
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Figure 1: Para-toluenesulfonamide (PTS) selectively accumulates in Tca-8113 cells. (a) Tca-8113 were treated with PTS or dimethyl sulfoxide (control vehicle) and then subjected to high-performance liquid chromatography. Peak 1: antipyrine (internal standard), peak 2: PTS. (b) Plot of intracellular PTS concentration after 2.5–80 μmol/l PTS treatment for different periods of time in Tca-8113 cells. (c) Plot of intracellular PTS concentration after 2.5–80 μmol/l PTS treatment for different periods in human gingival fibroblast (HGF) cells. (d) Comparative evaluation of the intracellular PTS concentration under the same treatment condition in Tca-8113 and HGF cells. The 10 min time-point was chosen to observe the difference in the intracellular PTS concentration between Tca-8113 and HGF cells. Results are representative of three independent experiments. *P<0.05.

Mentions: To examine the anticancer properties of PTS, we first measured the intracellular concentration of PTS in human tongue cancer Tca-8113 cells and gingival fibroblast HGF cells after PTS treatment. The HPLC method for the validation of PTS was performed with six calibration standards ranging from 10.0 to 320.0 μmol/l. The retention times of the internal standard and PTS were 2.0 and 12.6 min, respectively. As shown in Fig. 1a, PTS treatment increased the intracellular PTS concentration in Tca-8113 cells. We next assessed the accumulation of intracellular PTS after different doses and incubation periods of PTS treatment in Tca-8113 and HGF cells. Increased PTS treatment significantly induced a higher level of intracellular PTS accumulation in Tca-8113 and HGF cells. Prolonged PTS incubation also increased the intracellular PTS level. 80 μmol/l PTS treatment for 10 min induced maximal intracellular PTS accumulation in Tca-8113 and HGF cells (Fig. 1b and c). However, after 10 min of PTS treatment, intracellular PTS accumulation was significantly inhibited in HGF cells compared with that in Tca-8113 cells under the same PTS treatment condition (Fig. 1d). These results suggest that PTS selectively targets cancer cells, but exerts less effect on normal fibroblasts.


Para-toluenesulfonamide induces tongue squamous cell carcinoma cell death through disturbing lysosomal stability.

Liu Z, Liang C, Zhang Z, Pan J, Xia H, Zhong N, Li L - Anticancer Drugs (2015)

Para-toluenesulfonamide (PTS) selectively accumulates in Tca-8113 cells. (a) Tca-8113 were treated with PTS or dimethyl sulfoxide (control vehicle) and then subjected to high-performance liquid chromatography. Peak 1: antipyrine (internal standard), peak 2: PTS. (b) Plot of intracellular PTS concentration after 2.5–80 μmol/l PTS treatment for different periods of time in Tca-8113 cells. (c) Plot of intracellular PTS concentration after 2.5–80 μmol/l PTS treatment for different periods in human gingival fibroblast (HGF) cells. (d) Comparative evaluation of the intracellular PTS concentration under the same treatment condition in Tca-8113 and HGF cells. The 10 min time-point was chosen to observe the difference in the intracellular PTS concentration between Tca-8113 and HGF cells. Results are representative of three independent experiments. *P<0.05.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4588602&req=5

Figure 1: Para-toluenesulfonamide (PTS) selectively accumulates in Tca-8113 cells. (a) Tca-8113 were treated with PTS or dimethyl sulfoxide (control vehicle) and then subjected to high-performance liquid chromatography. Peak 1: antipyrine (internal standard), peak 2: PTS. (b) Plot of intracellular PTS concentration after 2.5–80 μmol/l PTS treatment for different periods of time in Tca-8113 cells. (c) Plot of intracellular PTS concentration after 2.5–80 μmol/l PTS treatment for different periods in human gingival fibroblast (HGF) cells. (d) Comparative evaluation of the intracellular PTS concentration under the same treatment condition in Tca-8113 and HGF cells. The 10 min time-point was chosen to observe the difference in the intracellular PTS concentration between Tca-8113 and HGF cells. Results are representative of three independent experiments. *P<0.05.
Mentions: To examine the anticancer properties of PTS, we first measured the intracellular concentration of PTS in human tongue cancer Tca-8113 cells and gingival fibroblast HGF cells after PTS treatment. The HPLC method for the validation of PTS was performed with six calibration standards ranging from 10.0 to 320.0 μmol/l. The retention times of the internal standard and PTS were 2.0 and 12.6 min, respectively. As shown in Fig. 1a, PTS treatment increased the intracellular PTS concentration in Tca-8113 cells. We next assessed the accumulation of intracellular PTS after different doses and incubation periods of PTS treatment in Tca-8113 and HGF cells. Increased PTS treatment significantly induced a higher level of intracellular PTS accumulation in Tca-8113 and HGF cells. Prolonged PTS incubation also increased the intracellular PTS level. 80 μmol/l PTS treatment for 10 min induced maximal intracellular PTS accumulation in Tca-8113 and HGF cells (Fig. 1b and c). However, after 10 min of PTS treatment, intracellular PTS accumulation was significantly inhibited in HGF cells compared with that in Tca-8113 cells under the same PTS treatment condition (Fig. 1d). These results suggest that PTS selectively targets cancer cells, but exerts less effect on normal fibroblasts.

Bottom Line: Para-toluenesulfonamide (PTS) has been implicated with anticancer effects against a variety of tumors.Next, the effects of PTS on cell viability, invasion, and cell death were determined.Lysosomal integrity assay and western blot showed that PTS increased lysosomal membrane permeabilization associated with activation of lysosomal cathepsin B.

View Article: PubMed Central - PubMed

Affiliation: aDepartment of Head and Neck Oncology bState Key Laboratory of Oral Diseases cDepartment of Oral and Maxillofacial Surgery, West China Hospital of Stomatology, Sichuan University, Chengdu dState Key Laboratory of Respiratory Disease, Guangzhou Institute of Respiratory Diseases, First Affiliated Hospital, Guangzhou Medical University, Guangzhou, China.

ABSTRACT
Para-toluenesulfonamide (PTS) has been implicated with anticancer effects against a variety of tumors. In the present study, we investigated the inhibitory effects of PTS on tongue squamous cell carcinoma (Tca-8113) and explored the lysosomal and mitochondrial changes after PTS treatment in vitro. High-performance liquid chromatography showed that PTS selectively accumulated in Tca-8113 cells with a relatively low concentration in normal fibroblasts. Next, the effects of PTS on cell viability, invasion, and cell death were determined. PTS significantly inhibited Tca-8113 cells' viability and invasive ability with increased cancer cell death. Flow cytometric analysis and the lactate dehydrogenase release assay showed that PTS induced cancer cell death by activating apoptosis and necrosis simultaneously. Morphological changes, such as cellular shrinkage, nuclear condensation as well as formation of apoptotic body and secondary lysosomes, were observed, indicating that PTS might induce cell death through disturbing lysosomal stability. Lysosomal integrity assay and western blot showed that PTS increased lysosomal membrane permeabilization associated with activation of lysosomal cathepsin B. Finally, PTS was shown to inhibit ATP biosynthesis and induce the release of mitochondrial cytochrome c. Therefore, our findings provide a novel insight into the use of PTS in cancer therapy.

Show MeSH
Related in: MedlinePlus