Enhancement of connexin30.3 expression in mouse embryonic stem cell line EB3 in response to cell–cell contacts

Tokunaga, Naruwa; Kishi, Ryota; Sasai, Tomoko; Saito, Mikako

doi:10.1007/s13577-018-00235-z

Cited by 2 publications

(1 citation statement)

References 27 publications

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“…Total RNA was prepared using ISOGEN II (Nippongene, Tokyo, Japan) according to the manufacturer’s instructions. According to a protocol described previously [ 6 ], the expression levels of Nanog , MMP2 , MMP9 , TGF-β1 mRNA were analyzed by quantitative (q)RT-PCR using a StepOnePlus™ Real-Time PCR System (Applied Biosystems) under the following conditions: 95 °C for 10 min, 45 cycles of a reaction set (95 °C denaturation for 15 s, 60 °C annealing for 1.0 min), and a reaction set for melt curve analysis (95 °C for 15 s and 60 °C for 1 min). Primer sets and expected product sizes of respective target RNAs are listed in a supplemental table (S1 Table).…”

Section: Methodsmentioning

confidence: 99%

Effect of Nanog overexpression on the metastatic potential of a mouse melanoma cell line B16-BL6

et al. 2021

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Nanog, a marker and regulator of the undifferentiated state in embryonic stem cells were anticipated to be an effective enhancer of cancer metastasis. We have developed a Nanog overexpressing mouse melanoma cell line B16-BL6 (BL6). BL6 was well recognized as a cell line with a high metastatic potential. In vitro tests revealed the enhancement of cell proliferation, wound healing activity, and matrix metalloproteinase 9 (MMP9) activity. Nanog-induced up- or down-regulated genes were comprehensively analyzed by transcriptome sequencing using Nanog+BL6 and wild-type BL6. Principally, up-regulated genes were involved in vesicle-aided glucose transport and oxidative phosphorylation, while down-regulated genes were associated with immunosuppression and apoptosis. A marked finding was that TGF-β1 was down-regulated, because TGF-β1 has been well discussed about its suppressive/progressive dual role in cancer. In vivo test showed that the number and volume of metastatic colonies of BL6 to lung were as high as 115 colonies/lung and 5.6 mm3/lung. Under this condition, Nanog overexpression caused a progressive effect (150 colonies/lung, p = 0.25; 9.2 mm3/lung, p = 0.13) rather than a suppressive effect on the metastasis. In this study, the effectiveness of Nanog overexpression in enhancing the metastatic potential of melanoma cell lines has been demonstrated for the first time.

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Section: Methodsmentioning

confidence: 99%