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. 2020 Nov 23;12(11):3482. doi: 10.3390/cancers12113482

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© 2020 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).

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The CR sublines exhibit highly invasive and cancer stemness properties. (A) Morphological appearance of CR cells compared to the parental (PT) cells, as examined by microscopy. (B) Cell invasion of CR and PT cells, as determined by a Matrigel invasion assay. The numbers of cells that had invaded through the Matrigel to the bottom chamber were determined after 16 h. (C) Tumorsphere formation by CR and PT cells as examined by tumorsphere assay, as described in the Methods. The ability of tumorsphere formation was quantified by the determination of relative sphere numbers. (D) The expression of stemness regulators (Nanog and OCT4) were evaluated in SAS-CR and CGHNC8-CR cells by immunoblot analysis and quantified by the determination of relative band density. The expression level of GAPDH was used as an internal control. (*: p ≤ 0.05, **: p ≤ 0.01, ***: p ≤ 0.001).

The CR sublines exhibit highly invasive and cancer stemness properties. (A) Morphological appearance of CR cells compared to the parental (PT) cells, as examined by microscopy. (B) Cell invasion of CR and PT cells, as determined by a Matrigel invasion assay. The numbers of cells that had invaded through the Matrigel to the bottom chamber were determined after 16 h. (C) Tumorsphere formation by CR and PT cells as examined by tumorsphere assay, as described in the Methods. The ability of tumorsphere formation was quantified by the determination of relative sphere numbers. (D) The expression of stemness regulators (Nanog and OCT4) were evaluated in SAS-CR and CGHNC8-CR cells by immunoblot analysis and quantified by the determination of relative band density. The expression level of GAPDH was used as an internal control. (*: p ≤ 0.05, **: p ≤ 0.01, ***: p ≤ 0.001).