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N-(2-hydroxyphenyl)acetamide (NA-2) and Temozolomide synergistically induce apoptosis in human glioblastoma cell line U87.

Hanif F, Perveen K, Jawed H, Ahmed A, Malhi SM, Jamall S, Simjee SU - Cancer Cell Int. (2014)

Bottom Line: The combine administration of NA-2 (0.33 mM) and temozolomide (0.1 mM) significantly enhanced the cell growth inhibition and apoptosis.Furthermore RT-PCR and imunocytochemistry data revealed that cooperative apoptosis induction was associated with increased ratio of Bax to Bcl-2 and active Caspase-3 expression.Our findings support that NA-2 possesses strong apoptotic activity and the combined administration of NA-2 and TMZ may be therapeutically exploited for the management of GBM.

View Article: PubMed Central - PubMed

Affiliation: Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, 75270 Pakistan.

ABSTRACT

Background: Despite the modern therapies available for treating glioblastoma multiforme (GBM), it is still a deadly disease. The development of new therapeutic strategies for the management of gliomas is therefore crucial. The present study is designed to analyze the therapeutic potentials of synthetic compound N-(2-hydroxyphenyl)acetamide (NA-2) in the treatment of GBM as a single agent or in combination with Temozolomide (TMZ) on glioblastoma cells.

Methods: MTT and TUNEL assays were used to detect the growth inhibitory effect and apoptotic activity of NA-2 alone and in combination with TMZ. Synergy was assessed using combination Index method. The expression of apoptosis related markers Bax, Bcl-2 and caspase-3 were assessed by RT-PCR, whereas, the active caspase-3 protein expression was determined using imunocytochemistry.

Results: Both NA-2 and TMZ inhibited the growth of U87 in a dose dependent manner. The combine administration of NA-2 (0.33 mM) and temozolomide (0.1 mM) significantly enhanced the cell growth inhibition and apoptosis. Furthermore RT-PCR and imunocytochemistry data revealed that cooperative apoptosis induction was associated with increased ratio of Bax to Bcl-2 and active Caspase-3 expression.

Conclusion: Our findings support that NA-2 possesses strong apoptotic activity and the combined administration of NA-2 and TMZ may be therapeutically exploited for the management of GBM.

No MeSH data available.


Related in: MedlinePlus

(A) Representative photomicrographs presenting increased apoptosis in cells exposed to temozolomide in combination with NA-2 revealed by TUNEL assay. U87 cells were treated with TMZ and NA-2 as single agent or in combination as indicated. After 24 hrs TUNEL method was used to detect apoptotic cells and photographs were taken under the microscope. Note that the dark brown stain is indicative of apoptosis and is strongest in cells treated with TMZ and NA-2 in combination. (B) Bar graph showing percentage of apoptotic cells by TUNEL assay and each bar represents mean ± S.E.M of three independent experiments. Significant difference between vehicle control and treated cells is indicated by **P <0.01 and ***P <0.001. Whereas the significant difference between the individual drugs NA-2 or TMZ and their combination treatment is indicated by $$$P <0.001, ###P <0.001 respectively.
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Fig3: (A) Representative photomicrographs presenting increased apoptosis in cells exposed to temozolomide in combination with NA-2 revealed by TUNEL assay. U87 cells were treated with TMZ and NA-2 as single agent or in combination as indicated. After 24 hrs TUNEL method was used to detect apoptotic cells and photographs were taken under the microscope. Note that the dark brown stain is indicative of apoptosis and is strongest in cells treated with TMZ and NA-2 in combination. (B) Bar graph showing percentage of apoptotic cells by TUNEL assay and each bar represents mean ± S.E.M of three independent experiments. Significant difference between vehicle control and treated cells is indicated by **P <0.01 and ***P <0.001. Whereas the significant difference between the individual drugs NA-2 or TMZ and their combination treatment is indicated by $$$P <0.001, ###P <0.001 respectively.

Mentions: U87 cells were treated with NA-2 (0.33 mM) and (TMZ 0.1 mM) alone and in combination for 24 hrs and apoptosis was detected by TUNEL assay. We found that combination treatment was more effective to induce apoptosis than the individual treatment of the drug (Figure 3A). Only 31.7% ±1.81% and 20.5% ±1.46% of apoptosis was observed when cells were treated with NA-2 and TMZ alone. Whereas, the percentage of apoptotic cells was dramatically increased to 53.6% ±1.40% in case of combination treatment (Figure 3B).Figure 3


N-(2-hydroxyphenyl)acetamide (NA-2) and Temozolomide synergistically induce apoptosis in human glioblastoma cell line U87.

Hanif F, Perveen K, Jawed H, Ahmed A, Malhi SM, Jamall S, Simjee SU - Cancer Cell Int. (2014)

(A) Representative photomicrographs presenting increased apoptosis in cells exposed to temozolomide in combination with NA-2 revealed by TUNEL assay. U87 cells were treated with TMZ and NA-2 as single agent or in combination as indicated. After 24 hrs TUNEL method was used to detect apoptotic cells and photographs were taken under the microscope. Note that the dark brown stain is indicative of apoptosis and is strongest in cells treated with TMZ and NA-2 in combination. (B) Bar graph showing percentage of apoptotic cells by TUNEL assay and each bar represents mean ± S.E.M of three independent experiments. Significant difference between vehicle control and treated cells is indicated by **P <0.01 and ***P <0.001. Whereas the significant difference between the individual drugs NA-2 or TMZ and their combination treatment is indicated by $$$P <0.001, ###P <0.001 respectively.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4319240&req=5

Fig3: (A) Representative photomicrographs presenting increased apoptosis in cells exposed to temozolomide in combination with NA-2 revealed by TUNEL assay. U87 cells were treated with TMZ and NA-2 as single agent or in combination as indicated. After 24 hrs TUNEL method was used to detect apoptotic cells and photographs were taken under the microscope. Note that the dark brown stain is indicative of apoptosis and is strongest in cells treated with TMZ and NA-2 in combination. (B) Bar graph showing percentage of apoptotic cells by TUNEL assay and each bar represents mean ± S.E.M of three independent experiments. Significant difference between vehicle control and treated cells is indicated by **P <0.01 and ***P <0.001. Whereas the significant difference between the individual drugs NA-2 or TMZ and their combination treatment is indicated by $$$P <0.001, ###P <0.001 respectively.
Mentions: U87 cells were treated with NA-2 (0.33 mM) and (TMZ 0.1 mM) alone and in combination for 24 hrs and apoptosis was detected by TUNEL assay. We found that combination treatment was more effective to induce apoptosis than the individual treatment of the drug (Figure 3A). Only 31.7% ±1.81% and 20.5% ±1.46% of apoptosis was observed when cells were treated with NA-2 and TMZ alone. Whereas, the percentage of apoptotic cells was dramatically increased to 53.6% ±1.40% in case of combination treatment (Figure 3B).Figure 3

Bottom Line: The combine administration of NA-2 (0.33 mM) and temozolomide (0.1 mM) significantly enhanced the cell growth inhibition and apoptosis.Furthermore RT-PCR and imunocytochemistry data revealed that cooperative apoptosis induction was associated with increased ratio of Bax to Bcl-2 and active Caspase-3 expression.Our findings support that NA-2 possesses strong apoptotic activity and the combined administration of NA-2 and TMZ may be therapeutically exploited for the management of GBM.

View Article: PubMed Central - PubMed

Affiliation: Dr. Panjwani Center for Molecular Medicine and Drug Research, International Center for Chemical and Biological Sciences, University of Karachi, Karachi, 75270 Pakistan.

ABSTRACT

Background: Despite the modern therapies available for treating glioblastoma multiforme (GBM), it is still a deadly disease. The development of new therapeutic strategies for the management of gliomas is therefore crucial. The present study is designed to analyze the therapeutic potentials of synthetic compound N-(2-hydroxyphenyl)acetamide (NA-2) in the treatment of GBM as a single agent or in combination with Temozolomide (TMZ) on glioblastoma cells.

Methods: MTT and TUNEL assays were used to detect the growth inhibitory effect and apoptotic activity of NA-2 alone and in combination with TMZ. Synergy was assessed using combination Index method. The expression of apoptosis related markers Bax, Bcl-2 and caspase-3 were assessed by RT-PCR, whereas, the active caspase-3 protein expression was determined using imunocytochemistry.

Results: Both NA-2 and TMZ inhibited the growth of U87 in a dose dependent manner. The combine administration of NA-2 (0.33 mM) and temozolomide (0.1 mM) significantly enhanced the cell growth inhibition and apoptosis. Furthermore RT-PCR and imunocytochemistry data revealed that cooperative apoptosis induction was associated with increased ratio of Bax to Bcl-2 and active Caspase-3 expression.

Conclusion: Our findings support that NA-2 possesses strong apoptotic activity and the combined administration of NA-2 and TMZ may be therapeutically exploited for the management of GBM.

No MeSH data available.


Related in: MedlinePlus