Small molecule tolfenamic acid and dietary spice curcumin treatment enhances antiproliferative effect in pancreatic cancer cells via suppressing Sp1, disrupting NF-kB translocation to nucleus and cell cycle phase distribution

Riyaz Mahammad Basha, Sarah F. Connelly, Umesh Tanaji Sankpal, Ganji Purnachandra Nagaraju, Hassaan Patel, Jamboor K. Vishwanatha, Sagar Shelake, Leslie Tabor-Simecka, Mamoru Shoji, Jerry Simecka, Bassel El-Rayes

Research output: Contribution to journalArticleResearchpeer-review

15 Citations (Scopus)

Abstract

Combination of dietary/herbal spice curcumin (Cur) and COX inhibitors has been tested for improving therapeutic efficacy in pancreatic cancer (PC). The objective of this study was to identify agent with low toxicity and COX-independent mechanism to induce PC cell growth inhibition when used along with Cur. Anticancer NSAID, tolfenamic acid (TA) and Cur combination were evaluated using PC cell lines. L3.6pl and MIA PaCa-2 cells were treated with Cur (5-25 μM) or TA (25-100 μM) or combination of Cur (7.5 μM) and TA (50 μM). Cell viability was measured at 24-72 h posttreatment using CellTiter-Glo kit. While both agents showed a steady/consistent effect, Cur + TA caused higher growth inhibition. Antiproliferative effect was compared with COX inhibitors, Ibuprofen and Celebrex. Cardiotoxicity was assessed using cordiomyocytes (H9C2). The expression of Sp proteins, survivin and apoptotic markers (western blot), caspase 3/7 (caspase-Glo kit), Annexin-V staining (flow cytometry), reactive oxygen species (ROS) and cell cycle phase distribution (flow cytometry) was measured. Cells were treated with TNF-α, and NF-kB translocation from cytoplasm to nucleus was evaluated (immunofluorescence). When compared to individual agents, combination of Cur + TA caused significant increase in apoptotic markers, ROS levels and inhibited NF-kB translocation to nucleus. TA caused cell cycle arrest in G0/G1, and the combination treatment showed mostly DNA synthesis phase arrest. These results suggest that combination of Cur + TA is less toxic and effectively enhance the therapeutic efficacy in PC cells via COX-independent mechanisms.

Original languageEnglish
Pages (from-to)77-87
Number of pages11
JournalJournal of Nutritional Biochemistry
Volume31
DOIs
StatePublished - 1 May 2016

Fingerprint

Spices
Curcumin
NF-kappa B
Pancreatic Neoplasms
Cell Cycle
Cells
Molecules
Flow cytometry
Celecoxib
Therapeutics
Reactive Oxygen Species
Flow Cytometry
Caspase 7
tolfenamic acid
Ibuprofen
Poisons
Annexin A5
Cell growth
Non-Steroidal Anti-Inflammatory Agents
Growth

Keywords

  • Combination treatment
  • Curcumin
  • Pancreatic cancer
  • Sp1
  • Tolfenamic acid

Cite this

@article{f12b7ca0ed964e88b44f1b7682cba4e9,
title = "Small molecule tolfenamic acid and dietary spice curcumin treatment enhances antiproliferative effect in pancreatic cancer cells via suppressing Sp1, disrupting NF-kB translocation to nucleus and cell cycle phase distribution",
abstract = "Combination of dietary/herbal spice curcumin (Cur) and COX inhibitors has been tested for improving therapeutic efficacy in pancreatic cancer (PC). The objective of this study was to identify agent with low toxicity and COX-independent mechanism to induce PC cell growth inhibition when used along with Cur. Anticancer NSAID, tolfenamic acid (TA) and Cur combination were evaluated using PC cell lines. L3.6pl and MIA PaCa-2 cells were treated with Cur (5-25 μM) or TA (25-100 μM) or combination of Cur (7.5 μM) and TA (50 μM). Cell viability was measured at 24-72 h posttreatment using CellTiter-Glo kit. While both agents showed a steady/consistent effect, Cur + TA caused higher growth inhibition. Antiproliferative effect was compared with COX inhibitors, Ibuprofen and Celebrex. Cardiotoxicity was assessed using cordiomyocytes (H9C2). The expression of Sp proteins, survivin and apoptotic markers (western blot), caspase 3/7 (caspase-Glo kit), Annexin-V staining (flow cytometry), reactive oxygen species (ROS) and cell cycle phase distribution (flow cytometry) was measured. Cells were treated with TNF-α, and NF-kB translocation from cytoplasm to nucleus was evaluated (immunofluorescence). When compared to individual agents, combination of Cur + TA caused significant increase in apoptotic markers, ROS levels and inhibited NF-kB translocation to nucleus. TA caused cell cycle arrest in G0/G1, and the combination treatment showed mostly DNA synthesis phase arrest. These results suggest that combination of Cur + TA is less toxic and effectively enhance the therapeutic efficacy in PC cells via COX-independent mechanisms.",
keywords = "Combination treatment, Curcumin, Pancreatic cancer, Sp1, Tolfenamic acid",
author = "Basha, {Riyaz Mahammad} and Connelly, {Sarah F.} and Sankpal, {Umesh Tanaji} and Nagaraju, {Ganji Purnachandra} and Hassaan Patel and Vishwanatha, {Jamboor K.} and Sagar Shelake and Leslie Tabor-Simecka and Mamoru Shoji and Jerry Simecka and Bassel El-Rayes",
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journal = "Journal of Nutritional Biochemistry",
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Small molecule tolfenamic acid and dietary spice curcumin treatment enhances antiproliferative effect in pancreatic cancer cells via suppressing Sp1, disrupting NF-kB translocation to nucleus and cell cycle phase distribution. / Basha, Riyaz Mahammad; Connelly, Sarah F.; Sankpal, Umesh Tanaji; Nagaraju, Ganji Purnachandra; Patel, Hassaan; Vishwanatha, Jamboor K.; Shelake, Sagar; Tabor-Simecka, Leslie; Shoji, Mamoru; Simecka, Jerry; El-Rayes, Bassel.

In: Journal of Nutritional Biochemistry, Vol. 31, 01.05.2016, p. 77-87.

Research output: Contribution to journalArticleResearchpeer-review

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T1 - Small molecule tolfenamic acid and dietary spice curcumin treatment enhances antiproliferative effect in pancreatic cancer cells via suppressing Sp1, disrupting NF-kB translocation to nucleus and cell cycle phase distribution

AU - Basha, Riyaz Mahammad

AU - Connelly, Sarah F.

AU - Sankpal, Umesh Tanaji

AU - Nagaraju, Ganji Purnachandra

AU - Patel, Hassaan

AU - Vishwanatha, Jamboor K.

AU - Shelake, Sagar

AU - Tabor-Simecka, Leslie

AU - Shoji, Mamoru

AU - Simecka, Jerry

AU - El-Rayes, Bassel

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AB - Combination of dietary/herbal spice curcumin (Cur) and COX inhibitors has been tested for improving therapeutic efficacy in pancreatic cancer (PC). The objective of this study was to identify agent with low toxicity and COX-independent mechanism to induce PC cell growth inhibition when used along with Cur. Anticancer NSAID, tolfenamic acid (TA) and Cur combination were evaluated using PC cell lines. L3.6pl and MIA PaCa-2 cells were treated with Cur (5-25 μM) or TA (25-100 μM) or combination of Cur (7.5 μM) and TA (50 μM). Cell viability was measured at 24-72 h posttreatment using CellTiter-Glo kit. While both agents showed a steady/consistent effect, Cur + TA caused higher growth inhibition. Antiproliferative effect was compared with COX inhibitors, Ibuprofen and Celebrex. Cardiotoxicity was assessed using cordiomyocytes (H9C2). The expression of Sp proteins, survivin and apoptotic markers (western blot), caspase 3/7 (caspase-Glo kit), Annexin-V staining (flow cytometry), reactive oxygen species (ROS) and cell cycle phase distribution (flow cytometry) was measured. Cells were treated with TNF-α, and NF-kB translocation from cytoplasm to nucleus was evaluated (immunofluorescence). When compared to individual agents, combination of Cur + TA caused significant increase in apoptotic markers, ROS levels and inhibited NF-kB translocation to nucleus. TA caused cell cycle arrest in G0/G1, and the combination treatment showed mostly DNA synthesis phase arrest. These results suggest that combination of Cur + TA is less toxic and effectively enhance the therapeutic efficacy in PC cells via COX-independent mechanisms.

KW - Combination treatment

KW - Curcumin

KW - Pancreatic cancer

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DO - 10.1016/j.jnutbio.2016.01.003

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