Journal: Journal of Oncology

Article Title: GEMIN6 Overexpression Correlates with the Low Immune Cell Infiltration and Poor Prognosis in Lung Adenocarcinoma

doi: 10.1155/2022/1930604

Figure Lengend Snippet: GEMIN6 expression in LUAD and other types of human cancers from TCGA data. (a) GEMIN6 expression in different tumor types; (b) the expression of GEMIN6 in LUAD and its paired adjacent tissues; (c) the expression of GEMIN6 in LUAD and normal tissues; (d) the association of GEMIN6 expression and T stages in LUAD; (e) the association of GEMIN6 expression and pathologic stages in LUAD; (f) the association of GEMIN6 expression and N stages in LUAD; (g) relative expression of GEMIN6 in human bronchial epithelial cells (BEAS-2B) and LUAD cell lines (A549, H1299, SK-MES-1, PC-9, and NCI-H23); (h) receiver operating characteristic analysis (ROC) of GEMIN6 in LUAD.

Article Snippet: The A549, H1299, SK-MES-1, PC-9, and NCI-H23 LUAD cell lines and the normal human bronchial epithelial cell line BEAS-2B were purchased from the iCell Bioscience Inc. (China).

Techniques: Expressing

Journal: Oncotarget

Article Title: Dual-strand tumor-suppressor microRNA-145 ( miR-145-5p and miR-145-3p ) coordinately targeted MTDH in lung squamous cell carcinoma

doi: 10.18632/oncotarget.12290

Figure Lengend Snippet: A. Expression levels of miR-145-5p and miR-145-3p in lung SCC clinical specimens and cell lines (SK-MES-1 and EBC-1) were determined by qRT-PCR. Data were normalized to RNU48 expression. B. Correlation of the expression levels of miR-145-5p and miR-145-3p . C. Cell growth was determined by XTT assays 72 h after transfection with 10 nM miR-145-5p or miR-145-3p . * P < 0.05. D. Cell migration activity was determined by wound healing assays. * P < 0.001. E. Cell invasion activity was determined using Matrigel invasion assays. * P < 0.001.

Article Snippet: Two human lung SCC cell lines (SK-MES-1 and EBC-1) were obtained from Japanese Cancer Research Resources Bank (JCRB) and the American Type Culture Collection (Manassas, VA, USA), respectively.

Techniques: Expressing, Quantitative RT-PCR, Transfection, Migration, Activity Assay

Journal: Oncotarget

Article Title: Dual-strand tumor-suppressor microRNA-145 ( miR-145-5p and miR-145-3p ) coordinately targeted MTDH in lung squamous cell carcinoma

doi: 10.18632/oncotarget.12290

Figure Lengend Snippet: A. Flow chart illustrates the strategy for analysis of miR-145-5p and miR-145-3p target genes in lung SCC cells. B. Identification of target genes of miR-145-5p or miR-145-3p . Expression levels of 7 mRNAs ( MTDH , EPN3 , TPD52 , CYP27B1 , LMAN1 , STAT1 and TXNDC12 ) were evaluated by qRT-PCR in EBC-1 cells 72 h after transfection with miR-145-5p or miR-145-3p . GUSB was used as an internal control. * P < 0.0001.

Article Snippet: Two human lung SCC cell lines (SK-MES-1 and EBC-1) were obtained from Japanese Cancer Research Resources Bank (JCRB) and the American Type Culture Collection (Manassas, VA, USA), respectively.

Techniques: Expressing, Quantitative RT-PCR, Transfection, Control

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: DAPL1 is activated by Np63 and GRα and regulates lipid metabolism

doi: 10.1007/s00109-025-02636-8

Figure Lengend Snippet: Dapl1 mRNA in LUSC is highly expressed ( A ) Three out of our four LUSC patients were upregulated in Dapl1 expression compared with adjacent normal (FPKM = fragments per kilobase per million reads). ( B ) Five LUAD have not detected or very low levels of Dapl1 expression in RNA-seq data. ( C ) Twenty-one Human LUSC and ( D ) twenty-one Human LUAD tissues were analyzed to determine Dapl1 expression level by PCR. Dapl1 was upregulated in 11 LUSC (52%), while no expression was observed in LUAD. ( E ) Expression levels of Dapl1 in TCGA (Cancer tissue and nearby normal tissue from the same patient) database, Dapl1 were upregulated in 36 among 51 LUSC (70.6%, mean RSEM 1142), but ( F ) Dapl1 was upregulated in 3 among 57 LUAD (5%, mean RSEM 13), RSEM (RNA-Seq by Expectation–Maximization) reports transcripts per million mapped reads (TPM). ( G ) Graph comparing Dapl1 expression in normal tissue and LUSC, and normal tissue and LUAD from GEPIA2 (GEPIA 2). ( H ) Colony formation assays were done by seeding HCC95 cells (1,000 cells/well) ( H-1 ) Graphing HCC95 colony formation assay results using Clono-counter, and ( I ) SKMES1 cells (1,000 cells/well) on 6 well plates. Dapl1 knockdown by siRNA dramatically reduced the colony formation. ( J ) Kaplan–Meier survival curve shows that the higher the Dapl1 expression, the worse the prognosis of lung cancer. Data was drawn from https://kmplot.com

Article Snippet: HCC95 and SKMES1 LUSC cells were purchased from Korean Cell Line Bank (KCLB, cellbank.snu.ac.kr, Seoul Korea).

Techniques: Expressing, RNA Sequencing, Colony Assay, Knockdown

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: DAPL1 is activated by Np63 and GRα and regulates lipid metabolism

doi: 10.1007/s00109-025-02636-8

Figure Lengend Snippet: Np63, GRα monomer are the transcription factors of DAPL1 and Dapl1 is increased in hypoxia depending on the situation. ( A ) When transcription factor Np63 was knocked out in MG-U74B skin cells, Dapl1 disappeared. (GDS1435/109381,109382, GEO Profiles, NCBI). ( B )( C ) Dapl1, Np63 expression in RNA-seq data of our 4 LUSC patients (cancer tissue and around normal tissue). ( D ) Among lung cancer cell lines, Np63, a transcription factor, was confirmed to be expressed in HCC95, which expresses Dapl1. (L132: Human cervix carcinoma. Originally derived from a human embryonic lung). ( E )( F ) When Transfecting the HCC95 cell with Np63siRNA, the expression of Np63, Dapl1 mRNA is identified by RT-PCR. ( G ) Estimated Np63 binding site in the DAPL1 promoter region based on a recent paper . ( H ) When osteosarcoma cells were transfected with GRα, GRα A, B, C, and D, Dapl1 increased in GRα, GRα A, B, and C over time, but did not increase in D (ID 56855360, GEO Profiles, NCBI). ( I ) The typical GRα dimer binding sequence and the monomer binding site at positions −797 to −810 bp of the DAPL1 promoter, for mutation experiments, mutate TGAA, ACGT. ( J ) Luciferase assay results, empty vector vs DAPL1 promoter vector, ( K ) DAPL1 promoter vector vs mutant vector, ( L ) DAPL1 promoter vector vs mutant vector-Dexamethasone 1uM, 3uM addition. ( M1,2 ) HCC95 and ( M3 , 4 ) HEK293T cells plated in 6 wells were grown under hypoxia conditions (N 2 94%, CO 2 5%, O 2 1%) incubator (Whitley H35 hypoxystation), after 18 h, 24 h, 70 h, it is compared with the control group grown under normal conditions by RT-PCR

Article Snippet: HCC95 and SKMES1 LUSC cells were purchased from Korean Cell Line Bank (KCLB, cellbank.snu.ac.kr, Seoul Korea).

Techniques: Expressing, RNA Sequencing, Derivative Assay, Reverse Transcription Polymerase Chain Reaction, Binding Assay, Transfection, Sequencing, Mutagenesis, Luciferase, Plasmid Preparation, Control

Journal: Journal of Molecular Medicine (Berlin, Germany)

Article Title: DAPL1 is activated by Np63 and GRα and regulates lipid metabolism

doi: 10.1007/s00109-025-02636-8

Figure Lengend Snippet: DAPL1 changes lipid synthesis enzymes. ( A ) Lipid synthesis pathways related to Fdft1, Pcyt1a, Sptlc1 genes. ( B ) Among the 10 genes, expression comparison of the finally selected Fdft1, Pcyt1a, and Sptlc1 genes in normal and adjacent cancer tissues of four LUSC patients. ( C ) When Dapl1 was overexpressed in HEK293T cells, changes in Fdft1, Pcyt1a, and Sptlc1 mRNA were confirmed by RT-PCR. ( D ) When Dapl1 was knockdown in HCC95 cells, changes in Dapl1, Fdft1, Pcyt1a, and Sptlc1 mRNA were confirmed by RT-PCR. ( E ) Schematic diagram of DAPL1 knockout using CRISPER-Cas9 system. 127 bp is deleted between exon2 and intron. The distance between two primers is 406 bp. Figure of genotyping results of generated mice. ( F ) In the kidney tissue of the DAPL1 KO male mouse, Dapl1, Fdft1, Pcyt1a, and Sptlc1 mRNA were confirmed by RT-PCR. ( G ) In 4 WT, 4 DAPL1 KO female mice’s eyes (high DAPL1 expression), Dapl1, Fdft1, Pcyt1a, and Sptlc1 mRNA were confirmed by RT-PCR. ( H ) In 3 WT, 3 DAPL1 KO female mice’s large intestines (no DAPL1 expression), Fdft1, Pcyt1a, and Sptlc1 mRNA were confirmed by RT-PCR. ( I ) After LC/MS analysis of 297 types of lipids in the eyes of male mice 3 WT and 3 DAPL1 KO, the principal component analysis (PCA) plot confirms the division into two groups. ( J ) In Partial least squares-discriminant analysis (PLS-DA), a supervised learning model, it was confirmed that the two groups were clearly divided. ( K ) From the importance scores, lipids of the TG class appeared to act as a major factor in model training. ( L ) In the Volcano plot, 18 types of TG & 2 types of DG have up-regulated (tendency shown in lipid class), TG 53:3, FA 20:4, LPE 18:0, PS 40:6, PS 34:1, PI 38:4, SM 44:2, Cer 40:2, LPC 18:0 were down-regulated (No tendency in lipid class) fold change > 1.5, -log(p) > 1.0. ( M ) In Heatmap analysis, an overall difference between the blue (Ho) and green (WT) groups could be confirmed

Article Snippet: HCC95 and SKMES1 LUSC cells were purchased from Korean Cell Line Bank (KCLB, cellbank.snu.ac.kr, Seoul Korea).

Techniques: Expressing, Comparison, Reverse Transcription Polymerase Chain Reaction, Knockdown, Knock-Out, Generated, Liquid Chromatography with Mass Spectroscopy