Induction of HT-29 colon cancer cells apoptosis by pyrogallol with growth inhibiting efficacy against drug-resistant Helicobacter pylori

Seemaisamy Revathi, Faruck L. Hakkim, Neelamegam R. Kumar, Hamid A. Bakshi, Luay Rashan, Mohammad Al-Balushi, Sidgi Syed Anwar Hasson, Muthukalingan Krishnan, Farideh Javid, Kayalvizhi Nagarajan

Research output: Contribution to journalArticle

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Abstract

Background: Colon cancer is the most aggressive form of cancers, that causes 0.5 million deaths per year around the globe. Targeting colon cancer by conventional therapeutic options elicits toxicity. Traditional medicines take a lead to alleviate the existing clinical challenges. Objective: To investigate antibacterial activity against Helicobacter Pylori and in vitro anti-colon cancer activity by Acacia nilotica extract (ACE) and its active constituent pyrogallol. Methods: Pyrogallol isolated from A. nilotica by column chromatography and HPLC and structure was elucidated by spectral analysis. Antibacterial activity was done by flow cytometry. Cytotoxicity was measured by MTT assay. Apoptotic morphology and nuclear fragmentation were assessed with AO/ethidium bromide and DAPI staining. DNA fragmentation was done by electrophoresis. Western blot used to analyze the molecular mechanism of apoptosis. Cell cycle arrest was determined using flow cytometry of propidium iodide stained cells. Cell migration was determined by wound healing assay. Results: ACE (20 µg/ml) and pyrogallol (10 µg/ml) treatment reduced the survival of H.pylori at 61% and 62%, respectively. MTT results show that HT-29 cells are more sensitive to pyrogallol with an IC 50 value of 35µg/ml compared to ACE. Pyrogallol treated HT-29 cells reached dead state i.e. late apoptotic state with severe nuclear fragmentation. Pyrogallol elicits dose dependent DNA fragmentation in HT-29 cells. Pyrogallol induced apoptosis by simultaneous down-regulation of Bcl-2 and up-regulation of BAX and cytochrome c. Pyrogallol arrested HT-29 cells in S and G2/M phase of cell cycle. Further pyrogallol exhibited marked antimetastatic potential by inhibiting the migration of HT-29 cells dose dependently. Conclusion: Both ACE and pyrogallol repressed the growth of H.pylori and as significant anti-colon cancer agent.

Original languageEnglish
Pages (from-to)1875-1884
Number of pages10
JournalAnti-Cancer Agents in Medicinal Chemistry
Volume18
Issue number13
DOIs
Publication statusPublished - Jan 1 2018

Fingerprint

Pyrogallol
Helicobacter pylori
Colonic Neoplasms
Apoptosis
HT29 Cells
Acacia
Growth
Pharmaceutical Preparations
Pylorus
DNA Fragmentation
Flow Cytometry
Ethidium
Propidium
G2 Phase
Traditional Medicine
Cell Cycle Checkpoints
Cytochromes c
Cell Division
Wound Healing
Cell Movement

Keywords

  • Acacia nilotica
  • Apoptosis
  • Colon cancer
  • Helicobacter pylori
  • HT-29
  • Pyrogallol

ASJC Scopus subject areas

  • Pharmacology
  • Drug Discovery

Cite this

Induction of HT-29 colon cancer cells apoptosis by pyrogallol with growth inhibiting efficacy against drug-resistant Helicobacter pylori. / Revathi, Seemaisamy; Hakkim, Faruck L.; Kumar, Neelamegam R.; Bakshi, Hamid A.; Rashan, Luay; Al-Balushi, Mohammad; Syed Anwar Hasson, Sidgi; Krishnan, Muthukalingan; Javid, Farideh; Nagarajan, Kayalvizhi.

In: Anti-Cancer Agents in Medicinal Chemistry, Vol. 18, No. 13, 01.01.2018, p. 1875-1884.

Research output: Contribution to journalArticle

Revathi, Seemaisamy ; Hakkim, Faruck L. ; Kumar, Neelamegam R. ; Bakshi, Hamid A. ; Rashan, Luay ; Al-Balushi, Mohammad ; Syed Anwar Hasson, Sidgi ; Krishnan, Muthukalingan ; Javid, Farideh ; Nagarajan, Kayalvizhi. / Induction of HT-29 colon cancer cells apoptosis by pyrogallol with growth inhibiting efficacy against drug-resistant Helicobacter pylori. In: Anti-Cancer Agents in Medicinal Chemistry. 2018 ; Vol. 18, No. 13. pp. 1875-1884.
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abstract = "Background: Colon cancer is the most aggressive form of cancers, that causes 0.5 million deaths per year around the globe. Targeting colon cancer by conventional therapeutic options elicits toxicity. Traditional medicines take a lead to alleviate the existing clinical challenges. Objective: To investigate antibacterial activity against Helicobacter Pylori and in vitro anti-colon cancer activity by Acacia nilotica extract (ACE) and its active constituent pyrogallol. Methods: Pyrogallol isolated from A. nilotica by column chromatography and HPLC and structure was elucidated by spectral analysis. Antibacterial activity was done by flow cytometry. Cytotoxicity was measured by MTT assay. Apoptotic morphology and nuclear fragmentation were assessed with AO/ethidium bromide and DAPI staining. DNA fragmentation was done by electrophoresis. Western blot used to analyze the molecular mechanism of apoptosis. Cell cycle arrest was determined using flow cytometry of propidium iodide stained cells. Cell migration was determined by wound healing assay. Results: ACE (20 µg/ml) and pyrogallol (10 µg/ml) treatment reduced the survival of H.pylori at 61{\%} and 62{\%}, respectively. MTT results show that HT-29 cells are more sensitive to pyrogallol with an IC 50 value of 35µg/ml compared to ACE. Pyrogallol treated HT-29 cells reached dead state i.e. late apoptotic state with severe nuclear fragmentation. Pyrogallol elicits dose dependent DNA fragmentation in HT-29 cells. Pyrogallol induced apoptosis by simultaneous down-regulation of Bcl-2 and up-regulation of BAX and cytochrome c. Pyrogallol arrested HT-29 cells in S and G2/M phase of cell cycle. Further pyrogallol exhibited marked antimetastatic potential by inhibiting the migration of HT-29 cells dose dependently. Conclusion: Both ACE and pyrogallol repressed the growth of H.pylori and as significant anti-colon cancer agent.",
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AU - Kumar, Neelamegam R.

AU - Bakshi, Hamid A.

AU - Rashan, Luay

AU - Al-Balushi, Mohammad

AU - Syed Anwar Hasson, Sidgi

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AU - Javid, Farideh

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N2 - Background: Colon cancer is the most aggressive form of cancers, that causes 0.5 million deaths per year around the globe. Targeting colon cancer by conventional therapeutic options elicits toxicity. Traditional medicines take a lead to alleviate the existing clinical challenges. Objective: To investigate antibacterial activity against Helicobacter Pylori and in vitro anti-colon cancer activity by Acacia nilotica extract (ACE) and its active constituent pyrogallol. Methods: Pyrogallol isolated from A. nilotica by column chromatography and HPLC and structure was elucidated by spectral analysis. Antibacterial activity was done by flow cytometry. Cytotoxicity was measured by MTT assay. Apoptotic morphology and nuclear fragmentation were assessed with AO/ethidium bromide and DAPI staining. DNA fragmentation was done by electrophoresis. Western blot used to analyze the molecular mechanism of apoptosis. Cell cycle arrest was determined using flow cytometry of propidium iodide stained cells. Cell migration was determined by wound healing assay. Results: ACE (20 µg/ml) and pyrogallol (10 µg/ml) treatment reduced the survival of H.pylori at 61% and 62%, respectively. MTT results show that HT-29 cells are more sensitive to pyrogallol with an IC 50 value of 35µg/ml compared to ACE. Pyrogallol treated HT-29 cells reached dead state i.e. late apoptotic state with severe nuclear fragmentation. Pyrogallol elicits dose dependent DNA fragmentation in HT-29 cells. Pyrogallol induced apoptosis by simultaneous down-regulation of Bcl-2 and up-regulation of BAX and cytochrome c. Pyrogallol arrested HT-29 cells in S and G2/M phase of cell cycle. Further pyrogallol exhibited marked antimetastatic potential by inhibiting the migration of HT-29 cells dose dependently. Conclusion: Both ACE and pyrogallol repressed the growth of H.pylori and as significant anti-colon cancer agent.

AB - Background: Colon cancer is the most aggressive form of cancers, that causes 0.5 million deaths per year around the globe. Targeting colon cancer by conventional therapeutic options elicits toxicity. Traditional medicines take a lead to alleviate the existing clinical challenges. Objective: To investigate antibacterial activity against Helicobacter Pylori and in vitro anti-colon cancer activity by Acacia nilotica extract (ACE) and its active constituent pyrogallol. Methods: Pyrogallol isolated from A. nilotica by column chromatography and HPLC and structure was elucidated by spectral analysis. Antibacterial activity was done by flow cytometry. Cytotoxicity was measured by MTT assay. Apoptotic morphology and nuclear fragmentation were assessed with AO/ethidium bromide and DAPI staining. DNA fragmentation was done by electrophoresis. Western blot used to analyze the molecular mechanism of apoptosis. Cell cycle arrest was determined using flow cytometry of propidium iodide stained cells. Cell migration was determined by wound healing assay. Results: ACE (20 µg/ml) and pyrogallol (10 µg/ml) treatment reduced the survival of H.pylori at 61% and 62%, respectively. MTT results show that HT-29 cells are more sensitive to pyrogallol with an IC 50 value of 35µg/ml compared to ACE. Pyrogallol treated HT-29 cells reached dead state i.e. late apoptotic state with severe nuclear fragmentation. Pyrogallol elicits dose dependent DNA fragmentation in HT-29 cells. Pyrogallol induced apoptosis by simultaneous down-regulation of Bcl-2 and up-regulation of BAX and cytochrome c. Pyrogallol arrested HT-29 cells in S and G2/M phase of cell cycle. Further pyrogallol exhibited marked antimetastatic potential by inhibiting the migration of HT-29 cells dose dependently. Conclusion: Both ACE and pyrogallol repressed the growth of H.pylori and as significant anti-colon cancer agent.

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