Journal: Frontiers in Genetics

Article Title: BTC as a Novel Biomarker Contributing to EMT via the PI3K-AKT Pathway in OSCC

doi: 10.3389/fgene.2022.875617

Figure Lengend Snippet: Association between the clinicopathological variables and BTC expression in 38 OSCC patients.

Article Snippet: The OSCC cell lines SCC4 and CAL27 were obtained from the Center for Molecular Medicine, Xiangya Hospital, Central South University (Changsha, China).

Techniques: Expressing, Virus, Infection

Journal: Frontiers in Genetics

Article Title: BTC as a Novel Biomarker Contributing to EMT via the PI3K-AKT Pathway in OSCC

doi: 10.3389/fgene.2022.875617

Figure Lengend Snippet: DEGs in OSCC. (A,B) Volcano plots were constructed using FC values and FDRs. The red points in the plot represent the overexpressed mRNAs, and the blue points indicate the downregulated mRNAs with statistical significance. (A) DEGs between normal and tumor tissues. (B) DEGs between metastasis-positive and metastasis-negative tumor tissues. (C) Hierarchical clustering analysis of mRNAs that were differentially expressed between metastasis-negative and metastasis-positive tissues. The normalized expression levels in the heatmaps are colored from blue to red in ascending order. (D) Multivariate Cox regression analysis according to gene expression. (E) Kaplan–Meier survival curves were performed to show the prognosis of patients with high and low expression of BTC through the analysis of the mRNA expression profile data of 260 OSCC tumor samples from TCGA database.

Article Snippet: The OSCC cell lines SCC4 and CAL27 were obtained from the Center for Molecular Medicine, Xiangya Hospital, Central South University (Changsha, China).

Techniques: Construct, Expressing, Gene Expression

Journal: Frontiers in Genetics

Article Title: BTC as a Novel Biomarker Contributing to EMT via the PI3K-AKT Pathway in OSCC

doi: 10.3389/fgene.2022.875617

Figure Lengend Snippet: BTC expression is correlated with clinicopathological parameters in OSCC patients. (A) Representative images of immunohistochemical staining and (B) immunoreactive score of BTC in human normal mucosa samples, OSCC tissue samples, and metastatic LN samples. The experiment was repeated three times independently. Results are shown as mean ± SD. T-test, n = 38. (C) mRNA expression of BTC in normal and tumor tissues was detected by RT-PCR. (D) Kaplan–Meier survival curves of 38 patients from our department. (E–L) Analysis of 330 OSCC samples from TCGA database and 32 pairs of OSCC samples selected from TCGA database showed the comparison of (E,F) BTC expressed in tumor tissues and normal tissues. Correlation analysis with BTC expression and (G) age, (H) histological grade, (I) sex, (J) T category, (K) tumor stage, and (L) LN metastasis status. * p < 0.05. ** p < 0.01. *** p < 0.001. **** p < 0.0001.

Article Snippet: The OSCC cell lines SCC4 and CAL27 were obtained from the Center for Molecular Medicine, Xiangya Hospital, Central South University (Changsha, China).

Techniques: Expressing, Immunohistochemical staining, Staining, Reverse Transcription Polymerase Chain Reaction, Comparison

Journal: Frontiers in Genetics

Article Title: BTC as a Novel Biomarker Contributing to EMT via the PI3K-AKT Pathway in OSCC

doi: 10.3389/fgene.2022.875617

Figure Lengend Snippet: Statistical analyses of clinicopathological features associated with survival in 38 OSCC patients with the multivariate Cox proportional hazards models.

Article Snippet: The OSCC cell lines SCC4 and CAL27 were obtained from the Center for Molecular Medicine, Xiangya Hospital, Central South University (Changsha, China).

Techniques: Expressing

Journal: Frontiers in Genetics

Article Title: BTC as a Novel Biomarker Contributing to EMT via the PI3K-AKT Pathway in OSCC

doi: 10.3389/fgene.2022.875617

Figure Lengend Snippet: Overexpression of BTC inhibits the proliferation, migration, and invasion of OSCC cell lines. (A) Cancer cell transfectants of the BTC-expressing vector and empty vector control were identified in SCC4 and CAL27 cells by Western blot. (B,C) Overexpression of BTC inhibited cell proliferation, as indicated by the CCK-8 assay, in CAL27 and SCC4 cells. (D) Wound healing assay showed that overexpression of BTC inhibited CAL27 and SCC4 cell migration. (E,F) Transwell assays showed that the migration and invasion abilities of CAL27 and SCC4 cells were impaired after overexpression of BTC.

Article Snippet: The OSCC cell lines SCC4 and CAL27 were obtained from the Center for Molecular Medicine, Xiangya Hospital, Central South University (Changsha, China).

Techniques: Over Expression, Migration, Expressing, Plasmid Preparation, Control, Western Blot, CCK-8 Assay, Wound Healing Assay

Journal: Frontiers in Genetics

Article Title: BTC as a Novel Biomarker Contributing to EMT via the PI3K-AKT Pathway in OSCC

doi: 10.3389/fgene.2022.875617

Figure Lengend Snippet: (A) Comparison of EMT scores in normal and tumor (in BTC low- and high-expression groups) tissue. (B) TCGA and GSEA showed highly regulated genes in patients with high-BTC expression versus those with low-BTC expression. (C) Heatmap of EMT marker expression in the BTC high- and low-expression groups. (D–F) Relationship between BTC and EMT markers, such as E-cadherin, N-cadherin, and vimentin. (G) PPI network analysis and Western blot analysis. The PPI network of the DEGs was constructed using STRING. The network nodes represent different proteins. The edges represent protein–protein associations, and the line thickness indicates the strength of the supporting data. (H) Protein expression level of EMT-related markers and the PI3K-AKT signaling pathway after overexpression of BTC in SCC4 and Cal27 cells.

Article Snippet: The OSCC cell lines SCC4 and CAL27 were obtained from the Center for Molecular Medicine, Xiangya Hospital, Central South University (Changsha, China).

Techniques: Comparison, Expressing, Marker, Western Blot, Construct, Over Expression

Journal: Human Molecular Genetics

Article Title: Permanent inactivation of Huntington's disease mutation by personalized allele-specific CRISPR/Cas9

doi: 10.1093/hmg/ddw286

Figure Lengend Snippet: Application of allele-specific dual gRNA-mediated CRISPR/Cas9 to independent cell lines. Different combinations of allele-specific gRNAs based on PAM-altering SNPs were tested on independent HD cell lines. We tested 1) an NPC line from an iPS cell line (CS97iHD-180n1) (27,28) derived from a Coriell fibroblast GM09197, and 2) an iPS cell line derived from a Coriell fibroblast GM04723. An NPC line of GM09197 carries hap.01 and hap.08 as expanded (CAG 180) and normal chromosome (CAG 18), respectively. An iPS line of GM04723 carries hap.03 and hap.08 as expanded (CAG 69) and normal chromosome (CAG15), respectively. Among other possible gRNA combinations, we tested 3 sets of gRNAs: (A) gRNA 1 and gRNA 2 (Fig. 2) to target hap.01 mutant allele in the NPC line, (B) gRNA 3 and gRNA 4 to target hap.01 mutant chromosome in the NPC line, and (C) gRNA 3 and gRNA 2 to target mutant hap.03 chromosome of an iPS cells. gRNA 3 and gRNA 4 depend on PAM sites on the mutant chromosomes, generated by SNPs rs2857935 and rs7659144, respectively (Supplementary Material, Table S4). After co-transfection of two CRISPR/Cas9 vectors expressing two gRNAs/puromycin selection, PCR assays were performed using sets of primers described in the method section. Expected size of excision for each experiment is shown under each gel picture. Based upon the PCR assay designs, each PCR products would be generated only if the large deletion occurs. EV represents an empty vector control transfection.

Article Snippet: An NPC cell line was generated from an iPS cell line (CS97iHD-180n1) by a STEMdiff protocol using Neural Induction Medium (STEMCELL Technologies).

Techniques: CRISPR, Derivative Assay, Mutagenesis, Generated, Cotransfection, Expressing, Selection, Plasmid Preparation, Control, Transfection