Journal: Scientific Reports

Article Title: Expression ratio of the TGFβ-inducible gene MYO10 is prognostic for overall survival of squamous cell lung cancer patients and predicts chemotherapy response

doi: 10.1038/s41598-018-27912-1

Figure Lengend Snippet: TGFβ treatment оf LUSC cells results in upregulation of migration- and actin cytoskeleton-related genes. ( A ) Non-supervised hierarchical clustering of z-scored differentially regulated 2323 genes (adjusted P -value < 0.01) between TGFβ-treated and untreated conditions. SK-MES1 cells were stimulated with 2 ng/ml TGFβ or left untreated. RNA was extracted and sequenced using HiSeq 4000. ( B ) Clusters of significantly upregulated GO cellular component gene sets between TGFβ-treated and untreated conditions. Significantly upregulated GO terms (adjusted P -value < 0.01) were visualized using REVIGO (allowed similarity 0.5). Thickness of connecting grey lines corresponds to the similarity of the GO terms. Only clusters that consist of at least two GO terms are displayed. ( C ) Volcano plot of differentially regulated genes between TGFβ-treated and untreated conditions. Fold change of averaged 8–48 h time points between both conditions is displayed. Only significantly regulated genes (adjusted P -value < 0.01) with a fold change of at least two are shown. Five most regulated genes from each cluster of upregulated GO cellular component gene sets are indicated with corresponding colors. Grey circles indicate differentially regulated genes that do not belong to any of the four clusters. ( D ) Time-resolved dynamics of top differentially regulated candidate genes from each of the clusters. Top five genes from each of the four clusters with the lowest adjusted P -values and fold change of at least two after normalization to untreated samples were selected as candidates. In case the same gene belonged to different clusters and satisfied the inclusion criteria, it was marked as belonging to both clusters. Single gene plots are shown in the Supplementary Fig. . ( E ) TCGA LUSC cohort RNA-Seq expression data of selected candidate genes sorted by frequency of mRNA upregulation. MYH9 , TGFB1 and MYO1E genes were additionally included.

Article Snippet: The human LUSC cell line SK-MES1 was purchased from ATCC and authenticated with a multiplexed human cell line authentication test (Multiplexion).

Techniques: Migration, RNA Sequencing, Expressing

Journal: Scientific Reports

Article Title: Expression ratio of the TGFβ-inducible gene MYO10 is prognostic for overall survival of squamous cell lung cancer patients and predicts chemotherapy response

doi: 10.1038/s41598-018-27912-1

Figure Lengend Snippet: MYO10 mRNA expression ratio is prognostic for overall survival of LUSC but not LUAD patients. ( A ) MYO10 mRNA expression ratio of tumor and adjacent non-tumor tissues in LUSC and LUAD patients. RNA was isolated from fresh-frozen resected tissues and MYO10 expression was measured using qRT-PCR. ( B ) Kaplan-Meier curves for overall survival using MYO10 mRNA expression ratio in LUSC and LUAD cohorts. Significance of difference between the two groups was tested using non-parametric Mann-Whitney U test. ( C ) Kaplan-Meier curves for adjuvant chemotherapy response in MYO10 low (left) and MYO10 high (right) patients. Significance of difference between the two groups was tested using non-parametric Mann-Whitney U test. ( D ) Differences of SNAI2 and TWIST1 expression depending on MYO10 mRNA expression ratio were tested by unpaired t -tests; *** P < 0.001; **** P < 0.0001. ( E ) MYO10 expression ratio in different stages of LUSC. Differences were tested by one-way ANOVA; * P ≤ 0.05. ( F ) Kaplan-Meier curve for the pathological stages of LUSC patients. Significance of difference between the two groups was tested using non-parametric Mann-Whitney U test. ( G ) Kaplan-Meier curves for lymph node status using MYO10 mRNA expression ratio. Significance of difference between the two groups was tested using non-parametric Mann-Whitney U test.

Article Snippet: The human LUSC cell line SK-MES1 was purchased from ATCC and authenticated with a multiplexed human cell line authentication test (Multiplexion).

Techniques: Expressing, Isolation, Quantitative RT-PCR, MANN-WHITNEY, Adjuvant

Journal: Scientific Reports

Article Title: Expression ratio of the TGFβ-inducible gene MYO10 is prognostic for overall survival of squamous cell lung cancer patients and predicts chemotherapy response

doi: 10.1038/s41598-018-27912-1

Figure Lengend Snippet: MYO10 mRNA expression ratio predicts outcome in lung cancer patient cohort.

Article Snippet: The human LUSC cell line SK-MES1 was purchased from ATCC and authenticated with a multiplexed human cell line authentication test (Multiplexion).

Techniques: Expressing

Journal: Scientific Reports

Article Title: Expression ratio of the TGFβ-inducible gene MYO10 is prognostic for overall survival of squamous cell lung cancer patients and predicts chemotherapy response

doi: 10.1038/s41598-018-27912-1

Figure Lengend Snippet: Differential expression of MYO10 is an independent predictor of survival in squamous cell carcinoma patients.

Article Snippet: The human LUSC cell line SK-MES1 was purchased from ATCC and authenticated with a multiplexed human cell line authentication test (Multiplexion).

Techniques: Quantitative Proteomics

Journal: Nature Communications

Article Title: c ircTP63 functions as a ceRNA to promote lung squamous cell carcinoma progression by upregulating FOXM1

doi: 10.1038/s41467-019-11162-4

Figure Lengend Snippet: Characterization of circTP63 in LUSC cells. a Genomic loci of circTP63 gene. circTP63 is produced at the TP63 gene (NM_003722.4) locus containing exons 10–11. The back-splice junction of circTP63 was identified by Sanger sequencing. b PCR analysis for circTP63 and its linear isoform TP63 in cDNA and genomic DNA (gDNA). c Northern blot analysis showed the size and abundance of circTP63 in one paired sample of LUSC tumorous tissue and corresponding adjacent nontumorous tissues. M: marker. d qRT-PCR for the abundance of circTP63 and TP63 in H1703 cells treated with Actinomycin D at the indicated time point. e Levels of circTP63 in the nuclear and cytoplasmic fractions of SW900 and H1703 cells. The error bars ( d , e ) represent s.d. ( n = 3)

Article Snippet: All of human LUSC cells (NCI-H2170, NCI-H1703, NCI-H226, NCI-H520, SW900, SK-MES-1, BEAS-2B, and HFL-1) were purchased from the American Type Culture Collection (ATCC) and were tested negative for mycoplasma contamination.

Techniques: Produced, Sequencing, Northern Blot, Marker, Quantitative RT-PCR

Journal: Nature Communications

Article Title: c ircTP63 functions as a ceRNA to promote lung squamous cell carcinoma progression by upregulating FOXM1

doi: 10.1038/s41467-019-11162-4

Figure Lengend Snippet: c ircTP63 promotes cell proliferation and tumor growth both in vitro and in vivo. a Expression levels of circTP63 and TP63 in SW900 and H1703 cells treated with circTP63 siRNA. b Expression levels of circTP63 and TP63 in H226 and H2170 cells after transduction with circTP63 lentivirus. c and d Cell proliferation analysis of LUSC cells with silencing or stably overexpressing circTP63 . e and f Cell cycle analysis of LUSC cells with silencing or stably overexpressing circTP63 . g The volume and weight of subcutaneous xenograft tumors of H2170 cells isolated from nude mice. h The volume and weight of subcutaneous xenograft tumors of H1703 cells isolated from nude mice; center line: median of data; Bounds of box: the second quartile to the third quartile; Whisker: minimum value to maximum value. The error bars a – h represent s.d. (in a – f , n = 3; in g and h , n = 6). * p < 0.05; ** p < 0.01; *** p < 0.001, two-tailed t -test. Source data are provided as a Source Data file

Article Snippet: All of human LUSC cells (NCI-H2170, NCI-H1703, NCI-H226, NCI-H520, SW900, SK-MES-1, BEAS-2B, and HFL-1) were purchased from the American Type Culture Collection (ATCC) and were tested negative for mycoplasma contamination.

Techniques: In Vitro, In Vivo, Expressing, Transduction, Stable Transfection, Cell Cycle Assay, Isolation, Whisker Assay, Two Tailed Test

Journal: Nature Communications

Article Title: c ircTP63 functions as a ceRNA to promote lung squamous cell carcinoma progression by upregulating FOXM1

doi: 10.1038/s41467-019-11162-4

Figure Lengend Snippet: c ircTP63 contributes to cell proliferation through targeting FOXM1. a Co-expression network of circTP63 with associated 25 mRNAs. A round node represents a protein-coding gene and the arrow node represents circTP63 ( hsa_circ_0068515 ). Lines between two nodes indicate interactions between two genes. Color represents the number of lines. b A heatmap shows mRNA levels of these 25 co-expression genes in the five paired LUSC samples of SBC Human ceRNA Array analysis. c Expression analysis for FOXM1 in additional 35 paired LUSC samples. d Correlation analysis revealed positive correlation between the levels of circTP63 and FOXM1 mRNA in the tumorous tissues of the 35 LUSC patients. ΔCt values were normalized according to β-actin . e The levels of circTP63 expression and FOXM1 protein in eight paired LUSC samples. f The mRNA and protein levels of FOXM1 in the LUSC cells with knockdown or overexpression of circTP63 . g Cell proliferation assay for H226 and H2170 cells with circTP63 overexpression and FOXM1 knockdown. h Cell proliferation assay for H1703 cells with circTP63 knockdown and FOXM1 overexpression. The error bars c , e – h represent s.d. (in c , n = 35; in e – h , n = 3). * p < 0.05; ** p < 0.01; *** p < 0.001, two-tailed t -test. Source data are provided as a Source Data file

Article Snippet: All of human LUSC cells (NCI-H2170, NCI-H1703, NCI-H226, NCI-H520, SW900, SK-MES-1, BEAS-2B, and HFL-1) were purchased from the American Type Culture Collection (ATCC) and were tested negative for mycoplasma contamination.

Techniques: Expressing, Knockdown, Over Expression, Proliferation Assay, Two Tailed Test

Journal: Nature Communications

Article Title: c ircTP63 functions as a ceRNA to promote lung squamous cell carcinoma progression by upregulating FOXM1

doi: 10.1038/s41467-019-11162-4

Figure Lengend Snippet: CENPA and CENPB are regulated by cicrTP63 through FOXM1. a The mRNA and protein levels of cell cycle-related genes in H226 and H2170 cells with circTP63 overexpression. b Expression changes of CENPA , CENPB , and CCNB1 after knockdown of FOXM1 in H2170 cells with circTP63 overexpression. c Cell proliferation assay for H226 and H2170 cells with circTP63 overexpression and joint knockdown of CENPA and CENPB . d Hypothesis diagram illustrates function and mechanism of circTP63 in LUSC progress. The error bars a – c represent s.d. ( n = 3). * p < 0.05; ** p < 0.01; *** p < 0.001, two-tailed t -test. Source data are provided as a Source Data file

Article Snippet: All of human LUSC cells (NCI-H2170, NCI-H1703, NCI-H226, NCI-H520, SW900, SK-MES-1, BEAS-2B, and HFL-1) were purchased from the American Type Culture Collection (ATCC) and were tested negative for mycoplasma contamination.

Techniques: Over Expression, Expressing, Knockdown, Proliferation Assay, Two Tailed Test

Journal: Molecules and Cells

Article Title: Extracellular Vesicles-Encapsulated miR-153-3p Potentiate the Survival and Invasion of Lung Adenocarcinoma

doi: 10.14348/molcells.2022.2221

Figure Lengend Snippet: (A) qRT-PCR analysis of miR-153-3p expression in LUSC cell lines (YTMLC-90, NCI-H266, and SK-MES-1) and LUAD cell lines (NCI-H1993, SW1271 and NCI-H522). (B) TEM of EVs derived from LUAD cell lines: EVs/ NCI-H1993, EVs/SW1271, and EVs/NCI-H522. (C) Nanoparticle tracking analysis of EVs/NCI-H1993, EVs/SW1271, and EVs/NCI-H522. (D) Western blot analysis of exosome markers (CD63, TSG101, Alix, and Hsp90) and non-exosome markers (GRP94 and Cytochrome) in LUAD cells lysate and separated EVs. (E) qRT-PCR analysis of miR-153-3p expression in EVs derived from LUSC cell lines (YTMLC-90, NCI-H266, and SK-MES-1) and LUAD cell lines (NCI-H1993, SW1271, and NCI-H522). (F) Heat map of the significant differentially expressed miRNAs extracted from selected lung carcinomas cell populations (small cell lung carcinoma, adenocarcinoma, and squamous cell carcinoma) compared with adjacent normal tissues (GSE74190). The data in the figures are represented the mean ± SD. ** P < 0.01.

Article Snippet: Human LUAD cell lines (NCI-H1993, SW1271, NCI-H522) and human lung squamous cell carcinoma (LUSC) cell lines (NCI-H226, YTMLC-90, SK-MES-1, NCI-H1915) were purchased from ATCC (USA).

Techniques: Quantitative RT-PCR, Expressing, Derivative Assay, Western Blot

Journal: Molecules and Cells

Article Title: Extracellular Vesicles-Encapsulated miR-153-3p Potentiate the Survival and Invasion of Lung Adenocarcinoma

doi: 10.14348/molcells.2022.2221

Figure Lengend Snippet: (A) Venn diagrams of potential downstream targets of miR-153-3p in two databases (Diana and StarBase). (B) The predicted miR-153-3p binding sites in the BANCR transcript. (C) Relative luciferase activity of reporter containing the 3’ UTR of BANCR upon transfection with miR-152-3p mimics or mimics NC in LUAD cells. (D) qRT-PCR analysis of BANCR expression in NCI-H1993, SW1271, and NCI-H522 cells. (E) Overall survival of LUAD and LUSC patients in different expression of BANCR from GEPIA database. (F) qRT-PCR analysis of BANCR expression in NCI-H1993, SW1271, and NCI-H522 cells after transfection with miR-152-3p inhibitor. (G) Western blot analysis of p-pi3k, pi3k, p-akt, akt in the treatment with inh-miR-153-3p, BANCR-KD, or inh-miR-153-3p+BANCR-KD. (H) The cell viability was assessed at 1, 2, and 3 days after treatment with inh-miR-153-3p, BANCR-KD, and inh-miR-153-3p+BANCR-KD. (I) The cell viability was assessed at 1, 2, and 3 days after treatment with inh-miR-153-3p, BANCR-KD, inh-miR-153-3p+LY294002, or BANCR-KD+ LY294002. (J) Western blot analysis of E-cadherin and vimentin in the treatment with inh-miR-153-3p, BANCR-KD, or inh-miR-153-3p+BANCR-KD. (K) The relationship between BANCR and miR-153-3p was detected by StarBase. The data in the figures are represented the mean ± SD. * P < 0.05; ** P < 0.01.

Article Snippet: Human LUAD cell lines (NCI-H1993, SW1271, NCI-H522) and human lung squamous cell carcinoma (LUSC) cell lines (NCI-H226, YTMLC-90, SK-MES-1, NCI-H1915) were purchased from ATCC (USA).

Techniques: Binding Assay, Luciferase, Activity Assay, Transfection, Quantitative RT-PCR, Expressing, Western Blot

Journal: Disease Markers

Article Title: circCNN2 Accelerates Cell Proliferation and Invasion in Lung Squamous Cell Carcinoma via Regulating miR-184/E2F1 and Activating MAPK Signaling Pathway

doi: 10.1155/2022/6329097

Figure Lengend Snippet: circCNN2 is highly expressed in LUSC cells. (a) RT-qPCR was used to examine the expression of circCNN2 in LUSC cell lines (SK-MES-1, H1703, H226, and H520) and the normal human bronchial epithelial Beas-2B cell line. (b) The specific convergent and divergent primers were, respectively, designed to amplify circCNN2, and gel electrophoresis with RT-qPCR was conducted to verify the loop structure of circCNN2. (c, d) RNase R and Actinomycin D (Act D) was used to treat H1703 and H226 cells and RT-qPCR was used to examine the expression of circCNN2 and CNN2. (e) FISH assay was carried out to identify the localization of circCNN2 in LUSC cells. ∗ P < 0.05, ∗∗ P < 0.01 compared to the control group.

Article Snippet: Human LUSC cell lines SK-MES-1 (HTB-67), H1703 (CRL-2073), H226 (CRL-5826), H520 (HTB-182), and the normal human bronchial epithelial Beas-2B cell line (CRL-9609) were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Quantitative RT-PCR, Expressing, Nucleic Acid Electrophoresis, Control

Journal: Disease Markers

Article Title: circCNN2 Accelerates Cell Proliferation and Invasion in Lung Squamous Cell Carcinoma via Regulating miR-184/E2F1 and Activating MAPK Signaling Pathway

doi: 10.1155/2022/6329097

Figure Lengend Snippet: circCNN2 promotes cell proliferation and invasion in LUSC. (a, b) CCK-8 and colony formation assays were conducted to evaluate the proliferative ability of H1703 and H226 cells after circCNN2 was silenced. (c) Western blot analysis was performed to examine the protein levels of PCNA and cyclin D1 in H1703 and H226 cells upon circCNN2 silencing. (d, e) Transwell assays were performed to analyze the effect of circCNN2 silencing on LUSC cell invasion. (f) Western blot analysis was used to examine the protein levels of invasion-related proteins MMP2 and MMP9 in LUSC cells. ∗∗ P < 0.01 compared to the sh-NC group.

Article Snippet: Human LUSC cell lines SK-MES-1 (HTB-67), H1703 (CRL-2073), H226 (CRL-5826), H520 (HTB-182), and the normal human bronchial epithelial Beas-2B cell line (CRL-9609) were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: CCK-8 Assay, Western Blot

Journal: Disease Markers

Article Title: circCNN2 Accelerates Cell Proliferation and Invasion in Lung Squamous Cell Carcinoma via Regulating miR-184/E2F1 and Activating MAPK Signaling Pathway

doi: 10.1155/2022/6329097

Figure Lengend Snippet: circCNN2 promotes LUSC tumor growth. (a) The tumor volume in H1703 cells upon circCNN2 knockdown was examined via RT-qPCR. (b) Tumor weight in H1703 cells upon circCNN2 silencing was measured via RT-qPCR. (c) The expression of Ki67, the nuclear antigen representing cell proliferation, was analyzed by immunohistochemistry. (d) Western blot analysis was utilized to examine the protein levels of proliferation-related proteins (PCNA and cyclin D1) and invasion-related proteins (MMP2 and MMP9) in different tumor groups. ∗∗ P < 0.01 compared to the sh-NC group.

Article Snippet: Human LUSC cell lines SK-MES-1 (HTB-67), H1703 (CRL-2073), H226 (CRL-5826), H520 (HTB-182), and the normal human bronchial epithelial Beas-2B cell line (CRL-9609) were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Knockdown, Quantitative RT-PCR, Expressing, Immunohistochemistry, Western Blot

Journal: Disease Markers

Article Title: circCNN2 Accelerates Cell Proliferation and Invasion in Lung Squamous Cell Carcinoma via Regulating miR-184/E2F1 and Activating MAPK Signaling Pathway

doi: 10.1155/2022/6329097

Figure Lengend Snippet: circCNN2 upregulates E2F1 expression via miR-184. (a) RT-qPCR was used to examine the expression of 8 candidate mRNAs (H2AFX, LSM4, ARHGAP39, CLUH, MDH2, E2F1, CDCA4, and TIMM13) upon circCNN2 knockdown. (b, c) RT-qPCR and western blot assays were conducted to test the expression and protein levels of E2F1 upon circCNN2 knockdown in H1703 and H226 cells. (d) RT-qPCR and western blot analysis were used to examine the expression and protein levels of E2F1 in LUSC cells. (e) RT-qPCR and western blot assays were conducted to detect the expression and protein levels of E2F1 upon miR-184 downregulation. (f) RNA pull down assay was carried out to verify the binding ability between miR-184 and E2F1 with Bio-NC as the control. (g) The binding ability between miR-184 and E2F1 was verified through luciferase report assay by testing the luciferase activity of LUSC cells transfected with wild-type or mutated E2F1 plasmids upon mimics NC and miR-184 mimics. (h) Rescue assays were conducted to verify the interaction among circCNN2, miR-184 and E2F1. ∗∗ P < 0.01 compared to the control group.

Article Snippet: Human LUSC cell lines SK-MES-1 (HTB-67), H1703 (CRL-2073), H226 (CRL-5826), H520 (HTB-182), and the normal human bronchial epithelial Beas-2B cell line (CRL-9609) were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Quantitative RT-PCR, Knockdown, Western Blot, Pull Down Assay, Binding Assay, Control, Luciferase, Activity Assay, Transfection

Journal: Disease Markers

Article Title: circCNN2 Accelerates Cell Proliferation and Invasion in Lung Squamous Cell Carcinoma via Regulating miR-184/E2F1 and Activating MAPK Signaling Pathway

doi: 10.1155/2022/6329097

Figure Lengend Snippet: circCNN2 activates MAPK signaling pathway by upregulating ASK1 expression via E2F1. (a) RT-qPCR and western blot assays were conducted to examine the expression and protein levels of ASK1 in LUSC cells. (b, c) RT-qPCR and western blot analysis were used to examine the expression and protein levels of ASK1 after E2F1 was knocked down in H1703 and H226 cells. (d, e) ChIP and DNA pull down assay were carried out to verify the binding ability between E2F1 and ASK1 promoter. (f) Luciferase reporter assay was conducted to detect the specific binding site between E2F1 and ASK1 promoter. (g) Western blot was conducted to detect the protein levels of p38 and p-p38 upon E2F1 silencing. (h) RT-qPCR was used to examine the expression of ASK1 in different transfection groups. (i) Western blot assay was conducted to examine the protein levels of p38, p-p38, and ASK1 under different transfection conditions. ∗∗ P < 0.01 compared to the control group.

Article Snippet: Human LUSC cell lines SK-MES-1 (HTB-67), H1703 (CRL-2073), H226 (CRL-5826), H520 (HTB-182), and the normal human bronchial epithelial Beas-2B cell line (CRL-9609) were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA).

Techniques: Expressing, Quantitative RT-PCR, Western Blot, Pull Down Assay, Binding Assay, Luciferase, Reporter Assay, Transfection, Control

Journal: Oncology Letters

Article Title: Clinical significance and effect of lncRNA BBOX1-AS1 on the proliferation and migration of lung squamous cell carcinoma

doi: 10.3892/ol.2021.13135

Figure Lengend Snippet: Flow chart of this study. LUSC, lung squamous cell carcinoma; lncRNA, long non-coding RNA; BBOX1-AS1, γ-butyrobetaine hydroxylase 1 antisense 1; CCK-8, Cell Counting Kit-8; SMD, standard mean deviation; SROC, summary receiver operating characteristic; TCGA, The Cancer Genome Atlas; GEO Gene Expression Omnibus; RT-qPCR, reverse transcription-quantitative PCR.

Article Snippet: Three human LUSC cell lines (NCI-H1703, NCI-H226 and SK-MES-1) and human bronchial epithelial cells (BEAS-2B) were obtained from the American Type Culture Collection.

Techniques: CCK-8 Assay, Cell Counting, Gene Expression, Quantitative RT-PCR, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Oncology Letters

Article Title: Clinical significance and effect of lncRNA BBOX1-AS1 on the proliferation and migration of lung squamous cell carcinoma

doi: 10.3892/ol.2021.13135

Figure Lengend Snippet: In-house microarray of lncRNAs in LUSC tissue. (A) Volcano plot; (B) the overlapping upregulated lncRNAs based on the datasets from the TCGA, GEO and ArrayExpress databases and the in-house microarray. TCGA, The Cancer Genome Atlas; GEO Gene Expression; LUSC, lung squamous cell carcinoma; lncRNA, long non-coding RNA; BBOX1-AS1, γ-butyrobetaine hydroxylase 1 antisense 1.

Article Snippet: Three human LUSC cell lines (NCI-H1703, NCI-H226 and SK-MES-1) and human bronchial epithelial cells (BEAS-2B) were obtained from the American Type Culture Collection.

Techniques: Microarray, Gene Expression

Journal: Oncology Letters

Article Title: Clinical significance and effect of lncRNA BBOX1-AS1 on the proliferation and migration of lung squamous cell carcinoma

doi: 10.3892/ol.2021.13135

Figure Lengend Snippet: Clinical significance of BBOX1-AS1 in LUSC based on RT-qPCR, as well as TCGA, GEO and ArrayExpress datasets. (A) Differential expression of BBOX1-AS1 between LUSC and normal lung tissue based on the TCGA dataset. (B) Differential expression of BBOX1-AS1 in the lymphatic metastasis group vs. no lymphatic metastasis group based on the TCGA dataset. (C) ROC curve of BBOX1-AS1 to discriminate LUSC from normal tissues. (D) Kaplan-Meier survival curves for BBOX1-AS1 expression in LUSC. (E) Differential expression of BBOX1-AS1 in LUSC cell lines based on RT-qPCR; ***P<0.001, ****P<0.0001 vs. BEAS-2B. (F-I) Differential expression of BBOX1-AS1 between LUSC and normal lung tissue in GEO and ArrayExpress datasets, including the (F) GSE19188 profile, (G) GSE30219 profile, (H) GSE103512 profile, and (I) E-MTAB-5231 profile. LUSC, lung squamous cell carcinoma; BBOX1-AS1, γ-butyrobetaine hydroxylase 1 antisense 1; ROC, receiver operating characteristic; TCGA, The Cancer Genome Atlas; GEO Gene Expression Omnibus; RT-qPCR, reverse transcription-quantitative PCR.

Article Snippet: Three human LUSC cell lines (NCI-H1703, NCI-H226 and SK-MES-1) and human bronchial epithelial cells (BEAS-2B) were obtained from the American Type Culture Collection.

Techniques: Quantitative RT-PCR, Quantitative Proteomics, Expressing, Gene Expression, Reverse Transcription, Real-time Polymerase Chain Reaction

Journal: Oncology Letters

Article Title: Clinical significance and effect of lncRNA BBOX1-AS1 on the proliferation and migration of lung squamous cell carcinoma

doi: 10.3892/ol.2021.13135

Figure Lengend Snippet: Expression of BBOX1-AS1 and the SROC curve for the included datasets. (A) Forest plot of datasets evaluating BBOX1-AS1 expression between the LUSC and normal control groups (random-effects model). (B) SROC curve for BBOX1-AS1 to discriminate LUSC. SMD, standard mean deviation; SROC, summary receiver operating characteristic; TCGA, The Cancer Genome Atlas; LUSC, lung squamous cell carcinoma; BBOX1-AS1, γ-butyrobetaine hydroxylase 1 antisense 1; AUC, area under the curve; SENS, sensitivity; SPEC, specificity.

Article Snippet: Three human LUSC cell lines (NCI-H1703, NCI-H226 and SK-MES-1) and human bronchial epithelial cells (BEAS-2B) were obtained from the American Type Culture Collection.

Techniques: Expressing, Control

Journal: Oncology Letters

Article Title: Clinical significance and effect of lncRNA BBOX1-AS1 on the proliferation and migration of lung squamous cell carcinoma

doi: 10.3892/ol.2021.13135

Figure Lengend Snippet: Images of lentivirus-transfected lung squamous cell carcinoma cells and the expression of BBOX1-AS1 in different groups after transfection. (A and B) Images of (A) SK-MES-1 and (B) NCI-H1703 cells transfected with lentivirus after 72 h (magnification, ×100 and ×200). B and G indicate transmitted light and fluorescence microscopy images, respectively. (C and D) Differential expression of BBOX1-AS1 in (C) SK-MES-1 and (D) NCI-H1703 cells after transfection determined by reverse transcription-quantitative PCR. *P<0.05. BBOX1-AS1, γ-butyrobetaine hydroxylase 1 antisense 1; NC, negative control; RNAi, RNA interference.

Article Snippet: Three human LUSC cell lines (NCI-H1703, NCI-H226 and SK-MES-1) and human bronchial epithelial cells (BEAS-2B) were obtained from the American Type Culture Collection.

Techniques: Transfection, Expressing, Fluorescence, Microscopy, Quantitative Proteomics, Reverse Transcription, Real-time Polymerase Chain Reaction, Negative Control

Journal: Oncology Letters

Article Title: Clinical significance and effect of lncRNA BBOX1-AS1 on the proliferation and migration of lung squamous cell carcinoma

doi: 10.3892/ol.2021.13135

Figure Lengend Snippet: Detection of the effect of γ-butyrobetaine hydroxylase 1 antisense 1 on the proliferation ability of lung squamous cell carcinoma cells by Cell Counting Kit-8 assay. (A) NCI-H1703 cells; (B) SK-MES-1 cells. *P<0.05, **P<0.01, ***P<0.001 for KD vs. NC at the same time-point. OD450, optical density at 450 nm; NC, negative control; KD, knockdown.

Article Snippet: Three human LUSC cell lines (NCI-H1703, NCI-H226 and SK-MES-1) and human bronchial epithelial cells (BEAS-2B) were obtained from the American Type Culture Collection.

Techniques: Cell Counting, Negative Control, Knockdown

Journal: Biomedicines

Article Title: Regulation of Oncogenic Targets by Tumor-Suppressive miR-150-3p in Lung Squamous Cell Carcinoma

doi: 10.3390/biomedicines9121883

Figure Lengend Snippet: Expression levels of miR-150-5p and miR-150-3p in LUSQ clinical specimens and LUSQ cell lines and effects of ectopic expression of miR-150-5p and miR-150-3p in LUSQ cells. ( A ) Expression levels of miR-150-5p and miR-150-3p were assessed in clinical specimens and cell lines (EBC-1 and SK-MES-1). Data were normalized to RNU48 . ( B ) XTT assays were performed after transfection of miR-150-5p or miR-150-3p to assess cell proliferation. ( C ) Cell cycle status after transfection of miR-150-5p or miR-150-3p was tested by flow cytometry. * p ≤ 0.0001.

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

Techniques: Expressing, Transfection, Flow Cytometry

Journal: Translational Cancer Research

Article Title: Nicotine-induced PD-L1 expression in lung squamous cell carcinoma is mediated by the α7-nAChR/STAT3 signaling pathway

doi: 10.21037/tcr-2024-2587

Figure Lengend Snippet: Nicotine-induced regulation of α7-nAChR, STAT3, and CD274 expression in LUSC cell lines. (A) Protein levels of α7-nAChR in BEAS-2B, H520, and SK-MES-1 cell lines. (B) qRT-PCR analysis of the effects of different concentrations of nicotine on the expression of CHRNA7 , STAT3 , and CD274 in LUSC cell lines. *, P<0.05; **, P<0.01. α7-nAChR, α7 nicotinic acetylcholine receptor; GAPDH, glyceraldehyde 3-phosphate dehydrogenase; LUSC, lung squamous cell carcinoma; mRNA, messenger RNA; qRT-PCR, quantitative real-time polymerase chain reaction; STAT3, signal transducer and activator of transcription 3.

Article Snippet: Three cell lines were used in this study: SK-MES-1 (Homo sapiens, human LUSC, Procell Life Science & Technology Co., Ltd., Wuhan, China, Cat#CL-0213), H520 (Homo sapiens, human LUSC, Procell Life Science & Technology Co., Ltd., Cat#CL-0402), and BEAS-2B (Homo sapiens, human normal bronchial epithelial cells, Procell Life Science & Technology Co., Ltd., Cat#CL-0496).

Techniques: Expressing, Quantitative RT-PCR, Real-time Polymerase Chain Reaction

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Chromobox Homologue 7 Acts as a Tumor Suppressor in Both Lung Adenocarcinoma and Lung Squamous Cell Carcinoma via Inhibiting ERK/MAPK Signaling Pathway

doi: 10.1155/2022/4952185

Figure Lengend Snippet: Chromobox homologue 7 (CBX7) was downregulated in lung adenocarcinoma (LUAD) and lung squamous cell carcinoma (LUSC). (a) Differentially expressed genes (DEGs) in LUAD were show in a volcano plot based on The Cancer Genome Atlas (TCGA) database ( https://www.cancer.gov/ ). (b) CBX7 expression in LUAD tissues and normal tissues was analyzed by the UALCAN ( http://ualcan.path.uab.edu/analysis.html ). (c) CBX7 expression in LUAD tissues of different clinicopathological stages was analyzed by the UALCAN. (d) Overall survival of LUAD patients with low and high CBX7 expression was analyzed by GEPIA ( http://gepia.cancer-pku.cn/ ). (e) CBX7 expression in LUSC tissues and normal tissues was analyzed by the UALCAN. (f) CBX7 expression in 6 tissues of different clinicopathological stages was analyzed by the UALCAN. (g) Relative mRNA expression of CBX7 in HPAEpiC, SK-MES-1, and A549 cells was measured by qRT-PCR. (h) Relative protein level of CBX7 in HPAEpiC, SK-MES-1, and A549 cells was measured by Western blotting. ∗ p < 0.05 and ∗∗ p < 0.01 vs. HPAEpiC.

Article Snippet: Human pulmonary alveolar epithelial cell line (HPAEpiC), LUAD cell line (A549), and LUSC cell line (SK-MES-1) were purchased from the American Type Culture Collection (ATCC; MD, USA).

Techniques: Expressing, Quantitative RT-PCR, Western Blot

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Chromobox Homologue 7 Acts as a Tumor Suppressor in Both Lung Adenocarcinoma and Lung Squamous Cell Carcinoma via Inhibiting ERK/MAPK Signaling Pathway

doi: 10.1155/2022/4952185

Figure Lengend Snippet: CBX7 overexpression inhibited the malignant progression of LUAD and LUSC cells. (a) The viability of A549 and SK-MES-1 cells was detected by CCK-8 assay. (b) The apoptosis of A549 and SK-MES-1 cells was detected by flow cytometry. (c) The migration of A549 and SK-MES-1 cells was measured by transwell assay. Scale bar = 50 µ m. (d) The invasion of A549 and SK-MES-1 cells was measured by transwell assay. Scale bar = 50 µ m. A549 and SK-MES-1 cells were transfected with lenti-NC or lenti-CBX7. ∗∗ p < 0.01 and ∗∗∗ p < 0.001 vs. lenti-NC.

Article Snippet: Human pulmonary alveolar epithelial cell line (HPAEpiC), LUAD cell line (A549), and LUSC cell line (SK-MES-1) were purchased from the American Type Culture Collection (ATCC; MD, USA).

Techniques: Over Expression, CCK-8 Assay, Flow Cytometry, Migration, Transwell Assay, Transfection

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Chromobox Homologue 7 Acts as a Tumor Suppressor in Both Lung Adenocarcinoma and Lung Squamous Cell Carcinoma via Inhibiting ERK/MAPK Signaling Pathway

doi: 10.1155/2022/4952185

Figure Lengend Snippet: CBX7 overexpression suppressed the ERK/MAPK signaling pathway in LUAD and LUSC cells. (a) Relative mRNA expression of CBX7, ERK1/2, and p38 in A549 and SK-MES-1 cells was detected by qRT-PCR. (b) Relative protein levels of CBX7, ERK1/2, p-ERK1/2, p38, and p-p38 in A549 and SK-MES-1 cells were detected by Western blotting. A549 and SK-MES-1 cells were transfected with lenti-NC or lenti-CBX7. ∗∗ p < 0.01 vs. lenti-NC.

Article Snippet: Human pulmonary alveolar epithelial cell line (HPAEpiC), LUAD cell line (A549), and LUSC cell line (SK-MES-1) were purchased from the American Type Culture Collection (ATCC; MD, USA).

Techniques: Over Expression, Expressing, Quantitative RT-PCR, Western Blot, Transfection

Journal: Evidence-based Complementary and Alternative Medicine : eCAM

Article Title: Chromobox Homologue 7 Acts as a Tumor Suppressor in Both Lung Adenocarcinoma and Lung Squamous Cell Carcinoma via Inhibiting ERK/MAPK Signaling Pathway

doi: 10.1155/2022/4952185

Figure Lengend Snippet: CBX7 overexpression inhibited the tumor growth of lung cancer via deactivating the ERK/MAPK pathway in vivo. (a) Representative images of the tumors generated by subcutaneously injection with A549 cells transfected with lenti-NC or lenti-CBX7. (b) Tumor volume and tumor weight. (c) The expression of CBX7 and Ki-67 in tumor tissues was determined by immunohistochemistry. Scale bar = 20 µ m. (d) The protein levels of CBX7, ERK1/2, p-ERK1/2, p38, and p-p38 in tumor tissues were measured by Western blotting. ∗∗ p < 0.01 vs. lenti-NC.

Article Snippet: Human pulmonary alveolar epithelial cell line (HPAEpiC), LUAD cell line (A549), and LUSC cell line (SK-MES-1) were purchased from the American Type Culture Collection (ATCC; MD, USA).

Techniques: Over Expression, In Vivo, Generated, Injection, Transfection, Expressing, Immunohistochemistry, Western Blot