DK1 induces apoptosis via mitochondria-dependent signaling pathway in human colon carcinoma cell linesIn vitro

Extensive research has been done in the search for innovative treatments against colon adenocarcinomas; however, the incidence rate of patients remains a major cause of cancer-related deaths in Malaysia. Natural bioactive compounds such as curcumin have been substantially studied as an alternative t...

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Main Authors: Hussin, Yazmin, Aziz, Muhammad Nazirul Mubin, Che Rahim, Nurul Fattin, Yeap, Swee Keong, Mohamad, Nurul Elyani, Masarudin, Mas Jaffri, Nordin, Noraini, Nik Abd Rahman, Nik Mohd Afizan, Yong, Chean Yeah, Akhtar, Muhammad Nadeem, Zamrus, Siti Noor Hajar, Mohammed Alitheen, Noorjahan Banu
Format: Article
Language:English
Published: MDPI 2018
Online Access:http://psasir.upm.edu.my/id/eprint/72322/
http://psasir.upm.edu.my/id/eprint/72322/1/DK1%20Induces%20Apoptosis.pdf
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author Hussin, Yazmin
Aziz, Muhammad Nazirul Mubin
Che Rahim, Nurul Fattin
Yeap, Swee Keong
Mohamad, Nurul Elyani
Masarudin, Mas Jaffri
Nordin, Noraini
Nik Abd Rahman, Nik Mohd Afizan
Yong, Chean Yeah
Akhtar, Muhammad Nadeem
Zamrus, Siti Noor Hajar
Mohammed Alitheen, Noorjahan Banu
author_facet Hussin, Yazmin
Aziz, Muhammad Nazirul Mubin
Che Rahim, Nurul Fattin
Yeap, Swee Keong
Mohamad, Nurul Elyani
Masarudin, Mas Jaffri
Nordin, Noraini
Nik Abd Rahman, Nik Mohd Afizan
Yong, Chean Yeah
Akhtar, Muhammad Nadeem
Zamrus, Siti Noor Hajar
Mohammed Alitheen, Noorjahan Banu
author_sort Hussin, Yazmin
building UPM Institutional Repository
collection Online Access
description Extensive research has been done in the search for innovative treatments against colon adenocarcinomas; however, the incidence rate of patients remains a major cause of cancer-related deaths in Malaysia. Natural bioactive compounds such as curcumin have been substantially studied as an alternative to anticancer drug therapies and have been surmised as a potent agent but, nevertheless, remain deficient due to its poor cellular uptake. Therefore, efforts now have shifted toward mimicking curcumin to synthesize novel compounds sharing similar effects. A synthetic analog, (Z)-3-hydroxy-1-(2-hydroxyphenyl)-3-phenylprop-2-ene-1-one (DK1), was recently synthesized and reported to confer improved bioavailability and selectivity toward human breast cancer cells. This study, therefore, aims to assess the anticancer mechanism of DK1 in relation to the induction of in vitro cell death in selected human colon cancer cell lines. Using the3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay, the cytotoxicity of DK1 towards HT29 and SW620 cell lines were investigated. Acridine orange/propidium iodide (AO/PI) dual-staining assay and flow cytometry analyses (cell cycle analysis, Annexin/V-FITC and JC-1 assays) were incorporated to determine the mode of cell death. To further determine the mechanism of cell death, quantitative real-time polymerase chain reaction (qRT-PCR) and proteome profiling were conducted. Results from this study suggest that DK1 induced changes in cell morphology, leading to a decrease in cell viability and subsequent induction of apoptosis. DK1 treatment inhibited cell viability and proliferation 48 h post treatment with IC50 values of 7.5 ± 1.6 µM for HT29 cells and 14.5 ± 4.3 µM for SW620 cells, causing cell cycle arrest with increased accumulation of cell populations at the sub-G0/G1phaseof 74% and 23%, respectively. Flow cytometry analyses showed that DK1 treatment in cancer cells induced apoptosis, as indicated by DNA fragmentation and depolarization of the mitochondrial membrane. qRT-PCR results show significant upregulation in the expression of caspase-9 in both HT29 and SW620 cell lines, further supporting that cell death induction by DK1 is via an intrinsic pathway. These outcomes, therefore, demonstrate DK1 as a potential anticancer agent for colon adenocarcinoma due to its anti-apoptotic attributes.
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spelling upm-723222020-05-04T07:49:04Z http://psasir.upm.edu.my/id/eprint/72322/ DK1 induces apoptosis via mitochondria-dependent signaling pathway in human colon carcinoma cell linesIn vitro Hussin, Yazmin Aziz, Muhammad Nazirul Mubin Che Rahim, Nurul Fattin Yeap, Swee Keong Mohamad, Nurul Elyani Masarudin, Mas Jaffri Nordin, Noraini Nik Abd Rahman, Nik Mohd Afizan Yong, Chean Yeah Akhtar, Muhammad Nadeem Zamrus, Siti Noor Hajar Mohammed Alitheen, Noorjahan Banu Extensive research has been done in the search for innovative treatments against colon adenocarcinomas; however, the incidence rate of patients remains a major cause of cancer-related deaths in Malaysia. Natural bioactive compounds such as curcumin have been substantially studied as an alternative to anticancer drug therapies and have been surmised as a potent agent but, nevertheless, remain deficient due to its poor cellular uptake. Therefore, efforts now have shifted toward mimicking curcumin to synthesize novel compounds sharing similar effects. A synthetic analog, (Z)-3-hydroxy-1-(2-hydroxyphenyl)-3-phenylprop-2-ene-1-one (DK1), was recently synthesized and reported to confer improved bioavailability and selectivity toward human breast cancer cells. This study, therefore, aims to assess the anticancer mechanism of DK1 in relation to the induction of in vitro cell death in selected human colon cancer cell lines. Using the3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide(MTT) assay, the cytotoxicity of DK1 towards HT29 and SW620 cell lines were investigated. Acridine orange/propidium iodide (AO/PI) dual-staining assay and flow cytometry analyses (cell cycle analysis, Annexin/V-FITC and JC-1 assays) were incorporated to determine the mode of cell death. To further determine the mechanism of cell death, quantitative real-time polymerase chain reaction (qRT-PCR) and proteome profiling were conducted. Results from this study suggest that DK1 induced changes in cell morphology, leading to a decrease in cell viability and subsequent induction of apoptosis. DK1 treatment inhibited cell viability and proliferation 48 h post treatment with IC50 values of 7.5 ± 1.6 µM for HT29 cells and 14.5 ± 4.3 µM for SW620 cells, causing cell cycle arrest with increased accumulation of cell populations at the sub-G0/G1phaseof 74% and 23%, respectively. Flow cytometry analyses showed that DK1 treatment in cancer cells induced apoptosis, as indicated by DNA fragmentation and depolarization of the mitochondrial membrane. qRT-PCR results show significant upregulation in the expression of caspase-9 in both HT29 and SW620 cell lines, further supporting that cell death induction by DK1 is via an intrinsic pathway. These outcomes, therefore, demonstrate DK1 as a potential anticancer agent for colon adenocarcinoma due to its anti-apoptotic attributes. MDPI 2018 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/72322/1/DK1%20Induces%20Apoptosis.pdf Hussin, Yazmin and Aziz, Muhammad Nazirul Mubin and Che Rahim, Nurul Fattin and Yeap, Swee Keong and Mohamad, Nurul Elyani and Masarudin, Mas Jaffri and Nordin, Noraini and Nik Abd Rahman, Nik Mohd Afizan and Yong, Chean Yeah and Akhtar, Muhammad Nadeem and Zamrus, Siti Noor Hajar and Mohammed Alitheen, Noorjahan Banu (2018) DK1 induces apoptosis via mitochondria-dependent signaling pathway in human colon carcinoma cell linesIn vitro. International Journal of Molecular Sciences, 19 (4). pp. 1-16. ISSN 1661-6596; ESSN: 1422-0067 https://www.mdpi.com/1422-0067/19/4/1151 10.3390/ijms19041151
spellingShingle Hussin, Yazmin
Aziz, Muhammad Nazirul Mubin
Che Rahim, Nurul Fattin
Yeap, Swee Keong
Mohamad, Nurul Elyani
Masarudin, Mas Jaffri
Nordin, Noraini
Nik Abd Rahman, Nik Mohd Afizan
Yong, Chean Yeah
Akhtar, Muhammad Nadeem
Zamrus, Siti Noor Hajar
Mohammed Alitheen, Noorjahan Banu
DK1 induces apoptosis via mitochondria-dependent signaling pathway in human colon carcinoma cell linesIn vitro
title DK1 induces apoptosis via mitochondria-dependent signaling pathway in human colon carcinoma cell linesIn vitro
title_full DK1 induces apoptosis via mitochondria-dependent signaling pathway in human colon carcinoma cell linesIn vitro
title_fullStr DK1 induces apoptosis via mitochondria-dependent signaling pathway in human colon carcinoma cell linesIn vitro
title_full_unstemmed DK1 induces apoptosis via mitochondria-dependent signaling pathway in human colon carcinoma cell linesIn vitro
title_short DK1 induces apoptosis via mitochondria-dependent signaling pathway in human colon carcinoma cell linesIn vitro
title_sort dk1 induces apoptosis via mitochondria-dependent signaling pathway in human colon carcinoma cell linesin vitro
url http://psasir.upm.edu.my/id/eprint/72322/
http://psasir.upm.edu.my/id/eprint/72322/
http://psasir.upm.edu.my/id/eprint/72322/
http://psasir.upm.edu.my/id/eprint/72322/1/DK1%20Induces%20Apoptosis.pdf