Phosphorylation of OCT4 Serine 236 Inhibits Germ Cell Tumor Growth by Inducing Differentiation

Kim, Dong-Keon; Song, Bomin; Han, Shuang; Jang, Hansol; Bae, Seung Hyun; Kim, Hee Yeon; Lee, Seon-Hyeong; Lee, Seungjin; Kim, Jong Kwang; Kim, Han Seong; Hong, Kyung Taek; Lee, Byung Il; Youn, Hong Duk; Kim, Soo Youl; Kang, Sang Won; Jang, Hyonchol

doi:10.3390/cancers12092601

Cited by 9 publications

(10 citation statements)

References 51 publications

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“…Cells were serially stained with the anti-SEZ6L2 antibody and goat anti-rabbit IgG (H+L) Cross-Adsorbed Secondary Antibody Alexa Fluor 488. Samples were analyzed at the Flow Cytometry Core Facility (National Cancer Center) using FACSVerse (BD Biosciences, San Jose, CA, USA), as described previously [ 25 ].…”

Section: Methodsmentioning

confidence: 99%

SEZ6L2 Is an Important Regulator of Drug-Resistant Cells and Tumor Spheroid Cells in Lung Adenocarcinoma

et al. 2020

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Many lung cancer deaths result from relapses in distant organs, such as the brain or bones, after standard chemotherapy. For cancer cells to spread to other organs, they must survive as circulating tumor cells (CTCs) in blood vessels. Thus, reducing distant recurrence after chemotherapy requires simultaneously inhibiting drug resistance and CTC survival. Here, we investigated the molecular pathways and genes that are commonly altered in drug-resistant lung cancer cells and lung tumor spheroid (TS) cells. First, RNA sequencing was performed in drug-resistant cells and TS cells originating from H460 and A549 lung cancer cells. Bioinformatic pathway analysis showed that cell cycle-related pathways were downregulated in drug-resistant cells, and cholesterol biosynthesis-related pathways were upregulated in TS cells. Seizure-related 6 homolog-like 2 (SEZ6L2) was selected as a gene that was commonly upregulated in both drug-resistant cells and TS cells, and that showed elevated expression in samples from lung adenocarcinoma patients. Second, the protein expression of SEZ6L2 was analyzed by flow cytometry. The proportions of SEZ6L2 positive cells among both drug-resistant cells and TS cells was increased. Finally, as SEZ6L2 is a transmembrane protein with an extracellular region, the function of SEZ6L2 was disrupted by treatment with an anti-SEZ6L2 antibody. Treatment with the anti-SEZ6L2 antibody reduced drug resistance and TS formation. Overall, our data showed that SEZ6L2 plays an important role in drug resistance and TS formation and may be a therapeutic target for reducing distant recurrence of lung adenocarcinoma.

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Section: Methodsmentioning

confidence: 99%

SEZ6L2 Is an Important Regulator of Drug-Resistant Cells and Tumor Spheroid Cells in Lung Adenocarcinoma

et al. 2020

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“…Next, 2TS22C cells were infected with retroviruses in the presence of 8 µg/ml polybrene (#H9268, Sigma). At 24 h after infection, Dox (1 μg/ml) was added to eliminate endogenous Sox2, and approximately 7 days later, ESC colonies were stained with alkaline phosphatase (AP) as described previously 25 .…”

Section: Retroviral Expression Of Sox2 Mutants and Self-renewal Assaymentioning

confidence: 99%

“…Animal experiments were performed as described previously 25 , with some modifications. E14 or E14-Sox2 TA/WT cells were dissociated and suspended in PBS supplemented with 50% Matrigel (#354234; BD Biosciences, USA) at a density of 1 × 10 7 cells/ml.…”

Section: Animal Experimentsmentioning

confidence: 99%

“…Teratomas were dissected and fixed in 4% paraformaldehyde overnight. After microdissection, sections were stained with hematoxylin and eosin (H&E) as described previously 25 at the Laboratory Animal Research Facility (National Cancer Center). H&E-stained slides were imaged at high resolution (200×) using the Phenochart 1.0.12 viewer from the Vectra Polaris 1.0 imaging system (PerkinElmer).…”

Section: Histological Analysis Of Teratomasmentioning

confidence: 99%

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O-GlcNAcylation of Sox2 at threonine 258 regulates the self-renewal and early cell fate of embryonic stem cells

Kim

Lee

et al. 2021

Exp Mol Med

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Sox2 is a core transcription factor in embryonic stem cells (ESCs), and O-GlcNAcylation is a type of post-translational modification of nuclear-cytoplasmic proteins. Although both factors play important roles in the maintenance and differentiation of ESCs and the serine 248 (S248) and threonine 258 (T258) residues of Sox2 are modified by O-GlcNAcylation, the function of Sox2 O-GlcNAcylation is unclear. Here, we show that O-GlcNAcylation of Sox2 at T258 regulates mouse ESC self-renewal and early cell fate. ESCs in which wild-type Sox2 was replaced with the Sox2 T258A mutant exhibited reduced self-renewal, whereas ESCs with the Sox2 S248A point mutation did not. ESCs with the Sox2 T258A mutation heterologously introduced using the CRISPR/Cas9 system, designated E14-Sox2TA/WT, also exhibited reduced self-renewal. RNA sequencing analysis under self-renewal conditions showed that upregulated expression of early differentiation genes, rather than a downregulated expression of self-renewal genes, was responsible for the reduced self-renewal of E14-Sox2TA/WT cells. There was a significant decrease in ectodermal tissue and a marked increase in cartilage tissue in E14-Sox2TA/WT-derived teratomas compared with normal E14 ESC-derived teratomas. RNA sequencing of teratomas revealed that genes related to brain development had generally downregulated expression in the E14-Sox2TA/WT-derived teratomas. Our findings using the Sox2 T258A mutant suggest that Sox2 T258 O-GlcNAc has a positive effect on ESC self-renewal and plays an important role in the proper development of ectodermal lineage cells. Overall, our study directly links O-GlcNAcylation and early cell fate decisions.

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“…The tumor sphere formation assay was carried out according to the previous method [26,27]. To put it simply, the transfected single cell suspension was inoculated in a 24-well plate with low adhesion for 2 weeks, and the spherical imaging was performed using Cytation 3 (Biotek, Winooski, VT, USA).…”

Section: Tumor Sphere Formation Assaymentioning

confidence: 99%

The expression of circ_0090049 in hepatocellular carcinoma and the molecular regulation mechanism of other biological functions

et al. 2021

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Hepatocellular carcinoma (HCC) is one of the most common malignant tumors in liver cancer. Circular RNA_0090049 (circ_0090049) has been shown to be involved in the advance of HCC. However, the interaction between circ_0090049 and microRNA (miRNA) in HCC has not been studied. Quantitative real-time PCR was used to detect the expression of related genes. Through detection of cell proliferation, migration, invasion, and rate of tumor sphere formation, the capping experiment was carried out to verify the regulatory relationship between miRNA and circ_0090049 or circ_0090049 and ubiquitin-conjugating enzyme E2 T (UBE2T). The expression of related proteins was detected by Western blotting. The interaction of miRNA with circ_0090049 or UBE2T was notarized by Dual-luciferase reporter assay. Xenotransplantation experiments confirmed the function of circ_0090049 in vivo. Circ_0090049 and UBE2T were upregulated in liver cancer. Silencing circ_0090049 reduced the proliferation, migration, invasion, and tumor spheroid formation rate of Huh7 and HCCLM3 cells. MiR-605-5p and miR-548c-3p were identified as targets of circ_0090049, and UBE2T was the target of miR-605-5p and miR-548c-3p. Anti-miR-605-5p, anti-miR-548c-3p or UBE2T overexpression restored the inhibitory effect of circ_0090049 knockdown on HCC cells. Animal experiments confirmed the antitumor effect of silence circ_0090049. Circ_0090049 regulates the expression of UBE2T by regulating miR-605-5p or miR-548c-3p, thereby promoting the development of HCC cells.

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Phosphorylation of OCT4 Serine 236 Inhibits Germ Cell Tumor Growth by Inducing Differentiation

Cited by 9 publications

References 51 publications

SEZ6L2 Is an Important Regulator of Drug-Resistant Cells and Tumor Spheroid Cells in Lung Adenocarcinoma

SEZ6L2 Is an Important Regulator of Drug-Resistant Cells and Tumor Spheroid Cells in Lung Adenocarcinoma

O-GlcNAcylation of Sox2 at threonine 258 regulates the self-renewal and early cell fate of embryonic stem cells

The expression of circ_0090049 in hepatocellular carcinoma and the molecular regulation mechanism of other biological functions

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