Open Access

Novel triple‑positive markers identified in human non‑small cell lung cancer cell line with chemotherapy-resistant and putative cancer stem cell characteristics

  • Authors:
    • Nazilah Abdul Satar
    • Kamal Shaik Fakiruddin
    • Moon Nian Lim
    • Pooi Ling Mok
    • Norashikin Zakaria
    • Noor Atiqah Fakharuzi
    • Ahmad Zuhairi Abd Rahman
    • Zubaidah Zakaria
    • Badrul Hisham Yahaya
    • Puteri Baharuddin
  • View Affiliations

  • Published online on: May 24, 2018     https://doi.org/10.3892/or.2018.6461
  • Pages: 669-681
  • Copyright: © Satar et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

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). Ιnitially, 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.
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August-2018
Volume 40 Issue 2

Print ISSN: 1021-335X
Online ISSN:1791-2431

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Spandidos Publications style
Satar NA, Fakiruddin KS, Lim MN, Mok PL, Zakaria N, Fakharuzi NA, Abd Rahman AZ, Zakaria Z, Yahaya BH, Baharuddin P, Baharuddin P, et al: Novel triple‑positive markers identified in human non‑small cell lung cancer cell line with chemotherapy-resistant and putative cancer stem cell characteristics. Oncol Rep 40: 669-681, 2018.
APA
Satar, N.A., Fakiruddin, K.S., Lim, M.N., Mok, P.L., Zakaria, N., Fakharuzi, N.A. ... Baharuddin, P. (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, 669-681. https://doi.org/10.3892/or.2018.6461
MLA
Satar, N. A., Fakiruddin, K. S., Lim, M. N., Mok, P. L., Zakaria, N., Fakharuzi, N. A., Abd Rahman, A. Z., Zakaria, Z., Yahaya, B. H., Baharuddin, P."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 (2018): 669-681.
Chicago
Satar, N. A., Fakiruddin, K. S., Lim, M. N., Mok, P. L., Zakaria, N., Fakharuzi, N. A., Abd Rahman, A. Z., Zakaria, Z., Yahaya, B. H., Baharuddin, P."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, no. 2 (2018): 669-681. https://doi.org/10.3892/or.2018.6461