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Identification of carcinogenic potential-associated molecular mechanisms in CD133+ A549 cells based on microRNA profiles

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Tumor Biology

Abstract

This study aimed to identify carcinogenic potential-related molecular mechanisms in cancer stem cells (CSCs) in lung cancer. CD133+ and CD133− subpopulations were sorted from A549 cells using magnetic-activated cell sorting. The abilities to form sphere and clone, proliferate, migrate, and invade were compared between CD133+ and CD133− cells, as well as drug sensitivity. Thereafter, microRNA (miRNA) profiles were performed to identify differentially expressed miRNAs between CD133+ and CD133− subpopulation. Following, bioinformatic methods were used to predict target genes for differentially expressed miRNAs and perform enrichment analysis. Furthermore, the mammalian target of rapamycin (mTOR) signaling pathways and CSC property-associated signaling pathways were explored and visualized in regulatory network among competitive endogenous RNA (ceRNA), miRNA, and target gene. CD133+ subpopulation showed greater oncogenic potential than CD133− subpopulation. In all, 14 differentially expressed miRNAs were obtained and enriched in 119 pathways, including five upregulated (hsa-miR-23b-3p, -23a-3p, -15b-5p, -24-3p, and -4734) and nine downregulated (hsa-miR-1246, -30b-5p, -5096, -6510-5p, has-miR-7110-5p, -7641, -3197, -7108-5p, and -6791-5p). For mTOR signaling pathway, eight differential miRNAs (hsa-miR-23b-3p, -23a-3p, -15b-5p, -24-3p, -4734, -1246, -7641, and -3197) and 39 target genes (e.g., AKT1, AKT2, PIK3CB, PIK3CG, PIK3R1, PIK3CA, and PIK3CD) were involved, as well as some ceRNAs. Besides, for CSC property-related signaling pathways, six miRNAs (hsa-miR-1246, -15b-5p, -30b-5p, -3197, -4734, and -7110-5p) were dramatically enriched in Hedgehog, Notch, and Wnt signaling pathways via regulating 108 target genes (e.g., DVL1, DVL3, WNT3A, and WNT5A). The mTOR and CSC property-associated signaling pathways may be important oncogenic molecular mechanisms in CD133+ A549 cells.

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Acknowledgments

This study was supported by “334” High-tech Talent Training Program of Nanjing Military Command and Natural Science Foundation of Zhejiang Province of China (LY12H16001).

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    Authors and Affiliations

    1. The First Affiliated Hospital of Wenzhou Medical University, Wenzhou, 325000, Zhejiang Province, People’s Republic of China

      Qing-yong Chen

    2. Department of Respiratory Disease, The 117th Hospital of PLA, Hangzhou, 310013, Zhejiang Province, People’s Republic of China

      Qing-yong Chen, De-min Jiao, Huizhen Hu, Jian Wang, Xiali Tang & Jun Chen

    3. Department of Oncology, The 117th Hospital of PLA, Hangzhou, 310013, Zhejiang Province, People’s Republic of China

      Ya Zhu & Li Yan

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    1. Qing-yong Chen
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    Correspondence to Qing-yong Chen or Li Yan.

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    Highlights

    1. CD133+ A549 cells showed greater carcinogenic potential than CD133− subpopulation.

    2. Totally, 14 differentially expressed miRNAs were identified in CD133+ A549 cells.

    3. Totally, 119 signaling pathways were enriched by 14 differentially expressed miRNAs.

    4. The mTOR signaling pathway was enriched by eight dysregulated miRNAs via 39 targets.

    5. Stem cell property-related pathways were enriched by six dysregulated miRNAs.

    Qing-yong Chen, De-min Jiao and Ya Zhu contributed equally to this work.

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    Supplementary Fig. 1

    CD133 expression checked by flow cytometry. a, CD133 expression of harvested cells before magnetic activated cell sorting; b, CD133 expression of CD133+ fractions after magnetic activated cell sorting. (GIF 15 kb)

    (TIFF 522 KB)

    Supplementary Fig. 2

    Network of differentially expressed miRNA and CD133 (PROM1). Red circle represents target gene, and green triangle represents differentially expressed miRNA. miRNA, microRNA. (GIF 46 kb)

    (TIFF 159 kb)

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    Chen, Qy., Jiao, Dm., Zhu, Y. et al. Identification of carcinogenic potential-associated molecular mechanisms in CD133+ A549 cells based on microRNA profiles. Tumor Biol. 37, 521–530 (2016). https://doi.org/10.1007/s13277-015-3675-9

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    • DOI: https://doi.org/10.1007/s13277-015-3675-9

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