The Effect of Novel 7-methyl-5-phenyl-pyrazolo[4,3-<i>e</i>]tetrazolo[4,5-<i>b</i>][1,2,4]triazine Sulfonamide Derivatives on Apoptosis and Autophagy in DLD-1 and HT-29 Colon Cancer Cells
The discovery of cytotoxic drugs is focused on designing a compound structure that directly affects cancer cells without an impact on normal cells. The mechanism of anticancer activity is mainly related with activation of apoptosis. However, recent scientific reports show that autophagy also plays a...
| Main Authors: | , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
MDPI AG
2020-07-01
|
| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/21/15/5221 |
| _version_ | 1797561548455018496 |
|---|---|
| author | Agnieszka Gornowicz Anna Szymanowska Mariusz Mojzych Krzysztof Bielawski Anna Bielawska |
| author_facet | Agnieszka Gornowicz Anna Szymanowska Mariusz Mojzych Krzysztof Bielawski Anna Bielawska |
| author_sort | Agnieszka Gornowicz |
| collection | DOAJ |
| description | The discovery of cytotoxic drugs is focused on designing a compound structure that directly affects cancer cells without an impact on normal cells. The mechanism of anticancer activity is mainly related with activation of apoptosis. However, recent scientific reports show that autophagy also plays a crucial role in cancer cell progression. Thus, the objective of this study was to synthesize 7-methyl-5-phenyl-pyrazolo[4,3-<i>e</i>]tetrazolo[4,5-<i>b</i>][1,2,4]triazine utilizing nucleophilic substitution reaction at the position N1. The biological activity of tested compounds was assessed in DLD-1 and HT-29 cell lines. The induction of apoptosis was confirmed by Annexin V binding assay and acridine orange/ethidium bromide staining. The loss of mitochondrial membrane potential and caspase-8 activity was estimated using cytometer flow analysis. The concentration of p53, LC3A, LC3B and beclin-1 was measured using the ELISA technique. Our study revealed that anticancer activity of 7-methyl-5-phenyl-pyrazolo[4,3-<i>e</i>]tetrazolo[4,5-<i>b</i>][1,2,4]triazine derivatives is related with initiation of apoptosis occur on the intrinsic pathway with mitochondrial membrane decrease and extrinsic with increase of activity of caspase-8. Moreover, a decrease in beclin-1, LC3A, and LC3B were observed in two cell lines after treatment with novel compounds. This study showed that novel 7-methyl-5-phenyl-pyrazolo[4,3-<i>e</i>]tetrazolo[4,5-<i>b</i>][1,2,4]triazine derivatives might be a potential strategy in colon cancer treatment. |
| first_indexed | 2024-03-10T18:16:29Z |
| format | Article |
| id | doaj.art-ee84de3ffee74516af2d3bf0127680a1 |
| institution | Directory Open Access Journal |
| issn | 1661-6596 1422-0067 |
| language | English |
| last_indexed | 2024-03-10T18:16:29Z |
| publishDate | 2020-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | International Journal of Molecular Sciences |
| spelling | doaj.art-ee84de3ffee74516af2d3bf0127680a12023-11-20T07:43:25ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672020-07-012115522110.3390/ijms21155221The Effect of Novel 7-methyl-5-phenyl-pyrazolo[4,3-<i>e</i>]tetrazolo[4,5-<i>b</i>][1,2,4]triazine Sulfonamide Derivatives on Apoptosis and Autophagy in DLD-1 and HT-29 Colon Cancer CellsAgnieszka Gornowicz0Anna Szymanowska1Mariusz Mojzych2Krzysztof Bielawski3Anna Bielawska4Department of Biotechnology, Medical University of Bialystok, 15-222 Bialystok, PolandDepartment of Biotechnology, Medical University of Bialystok, 15-222 Bialystok, PolandDepartment of Chemistry, Siedlce University of Natural Sciences and Humanities, 08-110 Siedlce, PolandDepartment of Synthesis and Technology of Drugs, Medical University of Bialystok, 15-222 Bialystok, PolandDepartment of Biotechnology, Medical University of Bialystok, 15-222 Bialystok, PolandThe discovery of cytotoxic drugs is focused on designing a compound structure that directly affects cancer cells without an impact on normal cells. The mechanism of anticancer activity is mainly related with activation of apoptosis. However, recent scientific reports show that autophagy also plays a crucial role in cancer cell progression. Thus, the objective of this study was to synthesize 7-methyl-5-phenyl-pyrazolo[4,3-<i>e</i>]tetrazolo[4,5-<i>b</i>][1,2,4]triazine utilizing nucleophilic substitution reaction at the position N1. The biological activity of tested compounds was assessed in DLD-1 and HT-29 cell lines. The induction of apoptosis was confirmed by Annexin V binding assay and acridine orange/ethidium bromide staining. The loss of mitochondrial membrane potential and caspase-8 activity was estimated using cytometer flow analysis. The concentration of p53, LC3A, LC3B and beclin-1 was measured using the ELISA technique. Our study revealed that anticancer activity of 7-methyl-5-phenyl-pyrazolo[4,3-<i>e</i>]tetrazolo[4,5-<i>b</i>][1,2,4]triazine derivatives is related with initiation of apoptosis occur on the intrinsic pathway with mitochondrial membrane decrease and extrinsic with increase of activity of caspase-8. Moreover, a decrease in beclin-1, LC3A, and LC3B were observed in two cell lines after treatment with novel compounds. This study showed that novel 7-methyl-5-phenyl-pyrazolo[4,3-<i>e</i>]tetrazolo[4,5-<i>b</i>][1,2,4]triazine derivatives might be a potential strategy in colon cancer treatment.https://www.mdpi.com/1422-0067/21/15/5221colorectal cancerapoptosisautophagyanticancer agentsroscovitine5-fluorouracil |
| spellingShingle | Agnieszka Gornowicz Anna Szymanowska Mariusz Mojzych Krzysztof Bielawski Anna Bielawska The Effect of Novel 7-methyl-5-phenyl-pyrazolo[4,3-<i>e</i>]tetrazolo[4,5-<i>b</i>][1,2,4]triazine Sulfonamide Derivatives on Apoptosis and Autophagy in DLD-1 and HT-29 Colon Cancer Cells International Journal of Molecular Sciences colorectal cancer apoptosis autophagy anticancer agents roscovitine 5-fluorouracil |
| title | The Effect of Novel 7-methyl-5-phenyl-pyrazolo[4,3-<i>e</i>]tetrazolo[4,5-<i>b</i>][1,2,4]triazine Sulfonamide Derivatives on Apoptosis and Autophagy in DLD-1 and HT-29 Colon Cancer Cells |
| title_full | The Effect of Novel 7-methyl-5-phenyl-pyrazolo[4,3-<i>e</i>]tetrazolo[4,5-<i>b</i>][1,2,4]triazine Sulfonamide Derivatives on Apoptosis and Autophagy in DLD-1 and HT-29 Colon Cancer Cells |
| title_fullStr | The Effect of Novel 7-methyl-5-phenyl-pyrazolo[4,3-<i>e</i>]tetrazolo[4,5-<i>b</i>][1,2,4]triazine Sulfonamide Derivatives on Apoptosis and Autophagy in DLD-1 and HT-29 Colon Cancer Cells |
| title_full_unstemmed | The Effect of Novel 7-methyl-5-phenyl-pyrazolo[4,3-<i>e</i>]tetrazolo[4,5-<i>b</i>][1,2,4]triazine Sulfonamide Derivatives on Apoptosis and Autophagy in DLD-1 and HT-29 Colon Cancer Cells |
| title_short | The Effect of Novel 7-methyl-5-phenyl-pyrazolo[4,3-<i>e</i>]tetrazolo[4,5-<i>b</i>][1,2,4]triazine Sulfonamide Derivatives on Apoptosis and Autophagy in DLD-1 and HT-29 Colon Cancer Cells |
| title_sort | effect of novel 7 methyl 5 phenyl pyrazolo 4 3 i e i tetrazolo 4 5 i b i 1 2 4 triazine sulfonamide derivatives on apoptosis and autophagy in dld 1 and ht 29 colon cancer cells |
| topic | colorectal cancer apoptosis autophagy anticancer agents roscovitine 5-fluorouracil |
| url | https://www.mdpi.com/1422-0067/21/15/5221 |
| work_keys_str_mv | AT agnieszkagornowicz theeffectofnovel7methyl5phenylpyrazolo43ieitetrazolo45ibi124triazinesulfonamidederivativesonapoptosisandautophagyindld1andht29coloncancercells AT annaszymanowska theeffectofnovel7methyl5phenylpyrazolo43ieitetrazolo45ibi124triazinesulfonamidederivativesonapoptosisandautophagyindld1andht29coloncancercells AT mariuszmojzych theeffectofnovel7methyl5phenylpyrazolo43ieitetrazolo45ibi124triazinesulfonamidederivativesonapoptosisandautophagyindld1andht29coloncancercells AT krzysztofbielawski theeffectofnovel7methyl5phenylpyrazolo43ieitetrazolo45ibi124triazinesulfonamidederivativesonapoptosisandautophagyindld1andht29coloncancercells AT annabielawska theeffectofnovel7methyl5phenylpyrazolo43ieitetrazolo45ibi124triazinesulfonamidederivativesonapoptosisandautophagyindld1andht29coloncancercells AT agnieszkagornowicz effectofnovel7methyl5phenylpyrazolo43ieitetrazolo45ibi124triazinesulfonamidederivativesonapoptosisandautophagyindld1andht29coloncancercells AT annaszymanowska effectofnovel7methyl5phenylpyrazolo43ieitetrazolo45ibi124triazinesulfonamidederivativesonapoptosisandautophagyindld1andht29coloncancercells AT mariuszmojzych effectofnovel7methyl5phenylpyrazolo43ieitetrazolo45ibi124triazinesulfonamidederivativesonapoptosisandautophagyindld1andht29coloncancercells AT krzysztofbielawski effectofnovel7methyl5phenylpyrazolo43ieitetrazolo45ibi124triazinesulfonamidederivativesonapoptosisandautophagyindld1andht29coloncancercells AT annabielawska effectofnovel7methyl5phenylpyrazolo43ieitetrazolo45ibi124triazinesulfonamidederivativesonapoptosisandautophagyindld1andht29coloncancercells |