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. 2021 May 3;11:581172. doi: 10.3389/fonc.2021.581172

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Copyright © 2021 Rasti, Madjd, Saeednejad Zanjani, Babashah, Abolhasani, Asgari and Mehrazma

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Clonogenicity and Sphere Formation Potential of Parental Cells (PCs) and Spheroid-Derived Cells (SDCs). (A) Sphere formation capacity of SDCs was significantly higher compared to PCs and was decreased in SDCs after transfection with SiSMAD4 (P < 0.01). The colony and sphere numbers were counted under a dissection microscope. (B) Three types of colonies termed holo, mero, and paraclones were identified during the colony formation assay in 3 cell populations, including PCs and SDCs before and after transfection with SiSMAD4. (C, D) The potential of clonogenicity was significantly higher in SDCs than PCs and was significantly decreased in SDCs after transfection with SiSMAD4. (E) The final results of cell invasion assay showed that SDCs were significantly more invasive (2.27-fold) than PCs, and invasive potential was significantly decreased (1.96-fold) in SDCs after transfection of PCs with SiSMAD4. Data are represented as mean ± SD (n = 3 each). **P < 0.01. (n = 3 each) and *P < 0.05.