Journal: Frontiers in Molecular Biosciences

Article Title: Knockdown of LncRNA PANDAR by CRISPR-dCas9 Decreases Proliferation and Increases Apoptosis in Oral Squamous Cell Carcinoma

doi: 10.3389/fmolb.2021.653787

Figure Lengend Snippet: The impact of PANDAR knockdown on proliferation and apoptosis of OSCC cells. ( A) The relative expression level of PANDAR in cancerous cell lines and normal cell line. ( B) The knockdown efficiency of specific sgRNAs targeting PANDAR was detected in OSCC cells by qRT-PCR. ( C–E) Cell proliferation assays (CCK-8 and EdU) and cell apoptosis assay (caspase-3 activity assay) were carried out to assess the functional role of PANDAR knockdown in OSCC. Data were represented as the mean values (±SD) of three independent experiments. * p < 0.05, significantly different from the control.

Article Snippet: To explore whether lncRNA PANDAR affects the biological activities of OSCC cells, we initially examined the expression level of PANDAR in OSCC cells (SAS, Cal27, SCC9, SCC15, and SCC4) and normal oral keratinocytes (NOKs).

Techniques: Knockdown, Expressing, Quantitative RT-PCR, CCK-8 Assay, Apoptosis Assay, Caspase-3 Activity Assay, Functional Assay, Control

Journal: Frontiers in Molecular Biosciences

Article Title: Knockdown of LncRNA PANDAR by CRISPR-dCas9 Decreases Proliferation and Increases Apoptosis in Oral Squamous Cell Carcinoma

doi: 10.3389/fmolb.2021.653787

Figure Lengend Snippet: SRSF7 acted as a RBP of PANDAR in OSCC. ( A) In line with the prediction of starBase, SRSF7 was found to be a RBP of PANDAR. (B) The expression level of SRSF7 in cancerous cells and normal cells. ( C,D) RIP and RNA pull-down assays were conducted for validating the interaction between SRSF7 and PANDAR in OSCC cells. ( E) PANDAR knockdown increased SRSF7 expression in OSCC cells. Data were represented as the mean values (±SD) of three independent experiments. * p < 0.05 and ** p < 0.01, significantly different from the control.

Article Snippet: To explore whether lncRNA PANDAR affects the biological activities of OSCC cells, we initially examined the expression level of PANDAR in OSCC cells (SAS, Cal27, SCC9, SCC15, and SCC4) and normal oral keratinocytes (NOKs).

Techniques: Expressing, Knockdown, Control

Journal: Frontiers in Molecular Biosciences

Article Title: Knockdown of LncRNA PANDAR by CRISPR-dCas9 Decreases Proliferation and Increases Apoptosis in Oral Squamous Cell Carcinoma

doi: 10.3389/fmolb.2021.653787

Figure Lengend Snippet: PANDAR affected PIM1 expression by regulating SRSF7 in OSCC. ( A) According to starBase, SRSF7 was identified as a RBP of PIM1. ( B) . The high expression level of PIM1 in OSCC cells. ( C,D) The interaction between SRSF7 and PIM1 was confirmed by RIP and RNA pull-down assays. ( E,F) The sgRNA targeting SRSF7 decreased SRSF7 and increased PIM1 expression. ( G,H) PIM1 expression level was reduced by sgRNA PANDAR#3, and this effect was reversed partially by sgRNA SRSF7 in OSCC cells. Data were represented as the mean values (±SD) of three independent experiments. * p < 0.05 and ** p < 0.01, significantly different from the control.

Article Snippet: To explore whether lncRNA PANDAR affects the biological activities of OSCC cells, we initially examined the expression level of PANDAR in OSCC cells (SAS, Cal27, SCC9, SCC15, and SCC4) and normal oral keratinocytes (NOKs).

Techniques: Expressing, Control

Journal: Frontiers in Molecular Biosciences

Article Title: Knockdown of LncRNA PANDAR by CRISPR-dCas9 Decreases Proliferation and Increases Apoptosis in Oral Squamous Cell Carcinoma

doi: 10.3389/fmolb.2021.653787

Figure Lengend Snippet: PANDAR modulated OSCC cell proliferation and apoptosis via SRSF7/PIM1 axis. ( A) The pcDNA3.1/PIM1 reversed the inhibitory effect on PIM1 mediated by knockdown of PANDAR. ( B–D) The inhibited proliferation and increased apoptosis of OSCC cells caused by PANDAR knockdown were recovered and weakened through overexpressing PIM1. Data were represented as the mean values (±SD) of three independent experiments. * p < 0.05, significantly different from the control.

Article Snippet: To explore whether lncRNA PANDAR affects the biological activities of OSCC cells, we initially examined the expression level of PANDAR in OSCC cells (SAS, Cal27, SCC9, SCC15, and SCC4) and normal oral keratinocytes (NOKs).

Techniques: Knockdown, Control

Journal: Cancer Gene Therapy

Article Title: Overexpression of angiopoietin 2 promotes the formation of oral squamous cell carcinoma by increasing epithelial–mesenchymal transition-induced angiogenesis

doi: 10.1038/cgt.2016.30

Figure Lengend Snippet: Angiopoietin 2 (ANG2) was upregulated in oral squamous cell carcinoma (OSCC) tissues. ( a ) Immunohistochemical and quantification analysis of ANG2 protein expression in normal liver tissue (up) and OSCC (bottom) samples using specific anti-ANG2 antibody. Representative photographs were obtained at × 200 magnification. ( b ) ANG2 mRNA expression levels in OSCC tissue samples and normal tissue samples (12 cases) were examined by real-time quantitative reverse transcriptase (qRT-PCR), respectively. ( c ) The expression of ANG2 in OSSC cell lines (TCA8113, cal27, SCC4, SCC15 and SCC25). GAPDH was used as an internal quantitative control. Three independent experiments were performed, and the data represent the means±s.d. * P <0.05.

Article Snippet: The human OSCC cell lines TCA8113, cal27, SCC4, SCC15 and SCC25 were obtained from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Immunohistochemical staining, Expressing, Reverse Transcription, Quantitative RT-PCR, Control

Journal: Cancer Gene Therapy

Article Title: Overexpression of angiopoietin 2 promotes the formation of oral squamous cell carcinoma by increasing epithelial–mesenchymal transition-induced angiogenesis

doi: 10.1038/cgt.2016.30

Figure Lengend Snippet: Effects of angiopoietin 2 (ANG2) on apoptosis and the cell cycle in oral squamous cell carcinoma (OSCC) cells. ( a ) Proliferation rates were determined by the MTS (3-(4, 5-dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium) assay on days 1–3 in TCA8113 mock cells or cells transfected with ANG2-encoded plasmids, ANG-targeted short interfering RNA (siRNA) or nonspecific (NC) siRNA. Before performing the MTS assays, cells were transfected with the indicated plasmids or siRNA for 24 h. Three independent experiments were performed, and the data represent the means±s.d. Treated and control groups of TCA8113 cells were used in subsequent experiments. ( b ) Representative dot plots showing fluorescence channel analysis of the treated and control groups of TCA8113 cells after dual staining with Annexin V and propidium iodide (PI) and analysis by flow cytometry. Columns represent the mean of three individual experiments; Bars, s.d. ( c ) Cell cycle distributions of the treated and control groups of TCA8113 cells were analyzed by flow cytometry. Data represent the mean±s.d. of three independent experiments.

Article Snippet: The human OSCC cell lines TCA8113, cal27, SCC4, SCC15 and SCC25 were obtained from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Transfection, Small Interfering RNA, Control, Fluorescence, Staining, Flow Cytometry

Journal: Cancer Gene Therapy

Article Title: Overexpression of angiopoietin 2 promotes the formation of oral squamous cell carcinoma by increasing epithelial–mesenchymal transition-induced angiogenesis

doi: 10.1038/cgt.2016.30

Figure Lengend Snippet: Overexpression of angiopoietin 2 (ANG2) increased the migration and invasion of TCA8113 cells by regulating epithelial–mesenchymal transition (EMT). ( a ) Cell migration and invasion were observed using a transwell chamber. ( b ) Western blot analysis of the cell proliferation-related proteins vimentin, Snail, Twist and E-cadherin in treated and control groups of TCA8113 cells. GAPDH served as a loading control. ( c ) Enzyme-linked immunosorbent assay (ELISA) was performed to analyze VEGF concentrations in cell-free supernatants. *Compared to Crtl or NC group. VEGF, vascular endothelial growth factor.

Article Snippet: The human OSCC cell lines TCA8113, cal27, SCC4, SCC15 and SCC25 were obtained from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Over Expression, Migration, Western Blot, Control, Enzyme-linked Immunosorbent Assay

Journal: Cancer Gene Therapy

Article Title: Overexpression of angiopoietin 2 promotes the formation of oral squamous cell carcinoma by increasing epithelial–mesenchymal transition-induced angiogenesis

doi: 10.1038/cgt.2016.30

Figure Lengend Snippet: Overexpression of angiopoietin 2 (ANG2) promoted tumorigenicity, increased angiogenesis and reduced apoptosis in nude mice. Subcutaneous injection of treated and control groups of TCA8113 cells into nude mice. ( a , b ) Tumor volume was examined every week. Three independent experiments were performed, and the data represent the means±s.d. * P <0.05. ( c ) Tumor tissues from nude mice 4 weeks after injection were subjected to terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) assays. Green TUNEL-positive cells are apoptotic cells with fragmented DNA, whereas blue Hoechst-positive cells represent all cells in this assay. Magnification is × 200. ( d ) Immunohistochemical and quantification analysis of CD31 protein expression in the treated and control groups of oral squamous cell carcinoma (OSCC) tissues with specific anti-CD31 antibody. Representative photographs were obtained at × 200 magnification. ( e ) Western blot analysis of tumor formation-related proteins including Bim, PUMA, Bcl-2, Bax, Cyclin D1 and PCNA in the treated and control groups of OSCC tissue samples. GAPDH served as a loading control.

Article Snippet: The human OSCC cell lines TCA8113, cal27, SCC4, SCC15 and SCC25 were obtained from the China Center for Type Culture Collection (Wuhan, China).

Techniques: Over Expression, Injection, Control, TUNEL Assay, Immunohistochemical staining, Expressing, Western Blot