Novel triple-positive markers identified in human non-small cell lung cancer cell line with chemotherapy-resistant and putative cancer stem cell characteristics
Through the specific identification and direct targeting of cancer stem cells (CSCs), it is believed that a better treatment efficacy of cancer may be achieved. Hence, the present study aimed to identify a CSC subpopulation from adenocarcinoma cells (A549) as a model of non-small cell lung cancer (N...
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| Format: | Article |
| Language: | English |
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Spandidos Publications
2018
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| Online Access: | http://psasir.upm.edu.my/id/eprint/75094/1/Novel%20triple-positive%20markers%20identified%20in%20human%20non-small%20cell%20lung%20cancer%20cell%20line%20with%20chemotherapy-resistant%20and%20putative%20cancer%20stem%20cell%20characteristics.pdf |
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| author | Abdul Satar, Nazilah Fakiruddin, Kamal Shaik Moon, Nian Lim Pooi, Ling Mok Zakaria, Norashikin Fakharuzi, Noor Atiqah Abd Rahman, Ahmad Zuhairi Zakaria, Zubaidah Yahaya, Badrul Hisham Baharuddin, Puteri |
| author_facet | Abdul Satar, Nazilah Fakiruddin, Kamal Shaik Moon, Nian Lim Pooi, Ling Mok Zakaria, Norashikin Fakharuzi, Noor Atiqah Abd Rahman, Ahmad Zuhairi Zakaria, Zubaidah Yahaya, Badrul Hisham Baharuddin, Puteri |
| author_sort | Abdul Satar, Nazilah |
| collection | UPM |
| description | Through the specific identification and direct targeting of cancer stem cells (CSCs), it is believed that a better treatment efficacy of cancer may be achieved. Hence, the present study aimed to identify a CSC subpopulation from adenocarcinoma cells (A549) as a model of non-small cell lung cancer (NSCLC). Initially, we sorted two subpopulations known as the triple-positive (EpCAM+/CD166+/CD44+) and triple-negative (EpCAM−/CD166−/CD44−) subpopulation using fluorescence-activated cell sorting (FACS). Sorted cells were subsequently evaluated for proliferation and chemotherapy-resistance using a viability assay and were further characterized for their clonal heterogeneity, self-renewal characteristics, cellular migration, alkaline dehydrogenase (ALDH) activity and the expression of stemness-related genes. According to our findings the triple-positive subpopulation revealed significantly higher (P<0.01) proliferation activity, exhibited better clonogenicity, was mostly comprised of holoclones and had markedly bigger (P<0.001) spheroid formation indicating a better self-renewal capacity. A relatively higher resistance to both 5-fluouracil and cisplatin with 80% expression of ALDH was observed in the triple-positive subpopulation, compared to only 67% detected in the triple-negative subpopulation indicated that high ALDH activity contributed to greater chemotherapy-resistance characteristics. Higher percentage of migrated cells was observed in the triple-positive subpopulation with 56% cellular migration being detected, compared to only 19% in the triple-negative subpopulation on day 2. This was similarly observed on day 3 in the triple-positive subpopulation with 36% higher cellular migration compared to the triple-negative subpopulation. Consistently, elevated levels of the stem cell genes such as REX1 and SSEA4 were also found in the triple-positive subpopulation indicating that the subpopulation displayed a strong characteristic of pluripotency. In conclusion, our study revealed that the triple-positive subpopulation demonstrated similar characteristics to CSCs compared to the triple-negative subpopulation. It also confirmed the feasibility of using the triple-positive (EpCAM+/CD166+/CD44+) marker as a novel candidate marker that may lead to the development of novel therapies targeting CSCs of NSCLC. |
| first_indexed | 2024-03-06T10:14:10Z |
| format | Article |
| id | upm.eprints-75094 |
| institution | Universiti Putra Malaysia |
| language | English |
| last_indexed | 2024-03-06T10:14:10Z |
| publishDate | 2018 |
| publisher | Spandidos Publications |
| record_format | dspace |
| spelling | upm.eprints-750942020-04-20T14:24:32Z http://psasir.upm.edu.my/id/eprint/75094/ Novel triple-positive markers identified in human non-small cell lung cancer cell line with chemotherapy-resistant and putative cancer stem cell characteristics Abdul Satar, Nazilah Fakiruddin, Kamal Shaik Moon, Nian Lim Pooi, Ling Mok Zakaria, Norashikin Fakharuzi, Noor Atiqah Abd Rahman, Ahmad Zuhairi Zakaria, Zubaidah Yahaya, Badrul Hisham Baharuddin, Puteri Through the specific identification and direct targeting of cancer stem cells (CSCs), it is believed that a better treatment efficacy of cancer may be achieved. Hence, the present study aimed to identify a CSC subpopulation from adenocarcinoma cells (A549) as a model of non-small cell lung cancer (NSCLC). Initially, we sorted two subpopulations known as the triple-positive (EpCAM+/CD166+/CD44+) and triple-negative (EpCAM−/CD166−/CD44−) subpopulation using fluorescence-activated cell sorting (FACS). Sorted cells were subsequently evaluated for proliferation and chemotherapy-resistance using a viability assay and were further characterized for their clonal heterogeneity, self-renewal characteristics, cellular migration, alkaline dehydrogenase (ALDH) activity and the expression of stemness-related genes. According to our findings the triple-positive subpopulation revealed significantly higher (P<0.01) proliferation activity, exhibited better clonogenicity, was mostly comprised of holoclones and had markedly bigger (P<0.001) spheroid formation indicating a better self-renewal capacity. A relatively higher resistance to both 5-fluouracil and cisplatin with 80% expression of ALDH was observed in the triple-positive subpopulation, compared to only 67% detected in the triple-negative subpopulation indicated that high ALDH activity contributed to greater chemotherapy-resistance characteristics. Higher percentage of migrated cells was observed in the triple-positive subpopulation with 56% cellular migration being detected, compared to only 19% in the triple-negative subpopulation on day 2. This was similarly observed on day 3 in the triple-positive subpopulation with 36% higher cellular migration compared to the triple-negative subpopulation. Consistently, elevated levels of the stem cell genes such as REX1 and SSEA4 were also found in the triple-positive subpopulation indicating that the subpopulation displayed a strong characteristic of pluripotency. In conclusion, our study revealed that the triple-positive subpopulation demonstrated similar characteristics to CSCs compared to the triple-negative subpopulation. It also confirmed the feasibility of using the triple-positive (EpCAM+/CD166+/CD44+) marker as a novel candidate marker that may lead to the development of novel therapies targeting CSCs of NSCLC. Spandidos Publications 2018 Article PeerReviewed text en http://psasir.upm.edu.my/id/eprint/75094/1/Novel%20triple-positive%20markers%20identified%20in%20human%20non-small%20cell%20lung%20cancer%20cell%20line%20with%20chemotherapy-resistant%20and%20putative%20cancer%20stem%20cell%20characteristics.pdf Abdul Satar, Nazilah and Fakiruddin, Kamal Shaik and Moon, Nian Lim and Pooi, Ling Mok and Zakaria, Norashikin and Fakharuzi, Noor Atiqah and Abd Rahman, Ahmad Zuhairi and Zakaria, Zubaidah and Yahaya, Badrul Hisham and Baharuddin, Puteri (2018) Novel triple-positive markers identified in human non-small cell lung cancer cell line with chemotherapy-resistant and putative cancer stem cell characteristics. Oncology Reports, 40 (2). 669 - 681. ISSN 1021-335X; ESSN: 1791-2431 10.3892/or.2018.6461 |
| spellingShingle | Abdul Satar, Nazilah Fakiruddin, Kamal Shaik Moon, Nian Lim Pooi, Ling Mok Zakaria, Norashikin Fakharuzi, Noor Atiqah Abd Rahman, Ahmad Zuhairi Zakaria, Zubaidah Yahaya, Badrul Hisham Baharuddin, Puteri Novel triple-positive markers identified in human non-small cell lung cancer cell line with chemotherapy-resistant and putative cancer stem cell characteristics |
| title | Novel triple-positive markers identified in human non-small cell lung cancer cell line with chemotherapy-resistant and putative cancer stem cell characteristics |
| title_full | Novel triple-positive markers identified in human non-small cell lung cancer cell line with chemotherapy-resistant and putative cancer stem cell characteristics |
| title_fullStr | Novel triple-positive markers identified in human non-small cell lung cancer cell line with chemotherapy-resistant and putative cancer stem cell characteristics |
| title_full_unstemmed | Novel triple-positive markers identified in human non-small cell lung cancer cell line with chemotherapy-resistant and putative cancer stem cell characteristics |
| title_short | Novel triple-positive markers identified in human non-small cell lung cancer cell line with chemotherapy-resistant and putative cancer stem cell characteristics |
| title_sort | novel triple positive markers identified in human non small cell lung cancer cell line with chemotherapy resistant and putative cancer stem cell characteristics |
| url | http://psasir.upm.edu.my/id/eprint/75094/1/Novel%20triple-positive%20markers%20identified%20in%20human%20non-small%20cell%20lung%20cancer%20cell%20line%20with%20chemotherapy-resistant%20and%20putative%20cancer%20stem%20cell%20characteristics.pdf |
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