Journal: Journal of Cellular Physiology

Article Title: Long non‐coding RNA SNHG15 promotes CDK14 expression via miR‐486 to accelerate non‐small cell lung cancer cells progression and metastasis

doi: 10.1002/jcp.26543

Figure Lengend Snippet: LncRNA SNHG15 was up‐regulated in NSCLC tissue and cells and indicated the poor prognosis. (a) RT‐PCR showed the SNHG15 expression in 35 cases of NSCLC tissue and adjacent noncancerous tissue. (b) SNHG15 expression in I–II stage and III stage of NSCLC tissue according to TNM pathological grades. (c) SNHG15 expression in NSCLC cell lines (A549, H460, SK‐MES‐1, and Calu‐3) and normal human bronchial epithelial cells (NHBE). (d) The distinction of NSCLC patients samples according to the SNHG15 expression (high/low). (e) Kaplan–Meier survival curves and log‐rank tests evaluated the survival rate of NSCLC patients with high or low SNHG15 expression. * p < 0.05, ** p < 0.01 presented significant difference

Article Snippet: The NSCLC cell lines (A549, H460, SK‐MES‐1, and Calu‐3) were purchased from the American Type Culture Collection (ATCC, Manassas, VA) and cultured in Dulbecco's Modified Eagle's medium (DMEM, Invitrogen, Carlsbad, CA).

Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing

Journal: Journal of Cellular Physiology

Article Title: Long non‐coding RNA SNHG15 promotes CDK14 expression via miR‐486 to accelerate non‐small cell lung cancer cells progression and metastasis

doi: 10.1002/jcp.26543

Figure Lengend Snippet: LncRNA SNHG15 silencing suppressed the proliferation of NSCLC cells in vitro. (a) RT‐PCR showed the SNHG15 expression in A549 and H460 cells transfected with three siRNA targeting SNHG15. (b,c) CCK‐8 assay showed the absorbance at 450 nm of A549 and H460 cells. (d,e) Colony formation assay showed the clone number in A549 and H460 cells respectively transfected with si‐SNHG15 and si‐NC. * p < 0.05, ** p < 0.01 presented significant difference

Article Snippet: The NSCLC cell lines (A549, H460, SK‐MES‐1, and Calu‐3) were purchased from the American Type Culture Collection (ATCC, Manassas, VA) and cultured in Dulbecco's Modified Eagle's medium (DMEM, Invitrogen, Carlsbad, CA).

Techniques: In Vitro, Reverse Transcription Polymerase Chain Reaction, Expressing, Transfection, CCK-8 Assay, Colony Assay

Journal: Journal of Cellular Physiology

Article Title: Long non‐coding RNA SNHG15 promotes CDK14 expression via miR‐486 to accelerate non‐small cell lung cancer cells progression and metastasis

doi: 10.1002/jcp.26543

Figure Lengend Snippet: LncRNA SNHG15 silencing induced cycle arrest at G0/G1 phase and suppressed the apoptosis of NSCLC cells. (a,b) Flow cytometry revealed the apoptosis of A549 and H460 cells transfected with si‐NC or si‐SNHG15. (c,d) Flow cytometry cycle analysis revealed the cells distribution of A549 and H460 cells transfected with si‐NC or si‐SNHG15. (e,f) Western blot showed the CDK14 protein expression in A549 and H460 cells. * p < 0.05, ** p < 0.01 presented significant difference

Article Snippet: The NSCLC cell lines (A549, H460, SK‐MES‐1, and Calu‐3) were purchased from the American Type Culture Collection (ATCC, Manassas, VA) and cultured in Dulbecco's Modified Eagle's medium (DMEM, Invitrogen, Carlsbad, CA).

Techniques: Flow Cytometry, Transfection, Western Blot, Expressing

Journal: Journal of Cellular Physiology

Article Title: Long non‐coding RNA SNHG15 promotes CDK14 expression via miR‐486 to accelerate non‐small cell lung cancer cells progression and metastasis

doi: 10.1002/jcp.26543

Figure Lengend Snippet: SNHG15 silencing inhibited the tumor growth of NSCLC cells in vivo. (a) The images of xenograft assay using A549 cells. (b) Tumor volume was measured after subcutaneous injection every 3 days. (c) Tumor weight was measured after mice sacrifice. (d) Blot images of CDK14 protein. (e) Quantitative data of CDK14 protein expression. Data were expressed as mean ± SD. * p < 0.05, ** p < 0.01 represents statistically difference

Article Snippet: The NSCLC cell lines (A549, H460, SK‐MES‐1, and Calu‐3) were purchased from the American Type Culture Collection (ATCC, Manassas, VA) and cultured in Dulbecco's Modified Eagle's medium (DMEM, Invitrogen, Carlsbad, CA).

Techniques: In Vivo, Xenograft Assay, Injection, Expressing

Journal: Journal of Cellular Physiology

Article Title: Long non‐coding RNA SNHG15 promotes CDK14 expression via miR‐486 to accelerate non‐small cell lung cancer cells progression and metastasis

doi: 10.1002/jcp.26543

Figure Lengend Snippet: SNHG15 positively regulated CDK14 expression via sponging miR‐486. (a) Schematic diagram showed the 3′‐UTR of SNHG15 wild‐type and mutant and miR‐486. (b) Luciferase reporter assay showed the luciferase activity of the combination of miR‐486 and SNHG15 wild‐type/mutant. (c) miR‐486 expression in NSCLC tissue samples compared with adjacent non‐tumor tissue. (d) Schematic diagram showed the binding within 3′‐UTR of CDK14 wild‐type and mutant and miR‐486. (e) Luciferase reporter assay showed the luciferase activity of the combination of miR‐486 and CDK14 wild‐type/mutant. (f) miR‐486 expression and CDK14 mRNA expression Iin A549 cells transfected with si‐SNHG15. (g) RT‐PCR showed CDK14 mRNA and SNHG15 expression levels in A549 cells transfected with miR‐486 inhibitor or miR‐486 mimics. (h,i) Western blot showed the CDK14 protein expression in A549 cells co‐transfected with miR‐486 inhibitor and si‐SNHG15. Data were expressed as mean ± SD. * p < 0.05, ** p < 0.01 represents statistically difference

Article Snippet: The NSCLC cell lines (A549, H460, SK‐MES‐1, and Calu‐3) were purchased from the American Type Culture Collection (ATCC, Manassas, VA) and cultured in Dulbecco's Modified Eagle's medium (DMEM, Invitrogen, Carlsbad, CA).

Techniques: Expressing, Mutagenesis, Luciferase, Reporter Assay, Activity Assay, Binding Assay, Transfection, Reverse Transcription Polymerase Chain Reaction, Western Blot

Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Hsa_circ_0129047 sponges miR-665 to attenuate lung adenocarcinoma progression by upregulating protein tyrosine phosphatase receptor type B

doi: 10.4196/kjpp.2023.27.2.131

Figure Lengend Snippet: (A) RT-qPCR analysis results of circ_0129047 in both LUAD and normal tissues. (B) RT-qPCR analysis of circ_0129047 in human bronchial epithelium (BEAS-2B) and LUAD cells (A549, H1975, PC9 and Calu-3). **p < 0.01 vs . BEAS-2B. (C) Loop structures of Circ_012904. (D) Subcellular localization of circ_0129047 in PC9 and Calu-3 cells. (E) Circ_012904 was resistant to RNase R digestion. **p < 0.01 vs . RNAse R. Values are presented as mean ± SD.

Article Snippet: Human bronchial epithelium (BEAS-2B, cat#: CRL-9609), LUAD cells (A549, cat#: CRM-CCL-185 derived from the lung tissue of a white, 58-year-old male with LUAD; H1975, CRL-5908 derived from the lung tissue of a nonsmoking female with LUAD; and Calu-3, cat#: HTB-55, derived from the lung tissue of a white, 25-year-old male with LUAD) were obtained from the American Type Culture Collection (ATCC), and PC9 (cat#: 90071810, derived from the lung tissue of an undifferentiated patient with LUAD) was obtained from Sigma-Aldrich.

Techniques: Quantitative RT-PCR

Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Hsa_circ_0129047 sponges miR-665 to attenuate lung adenocarcinoma progression by upregulating protein tyrosine phosphatase receptor type B

doi: 10.4196/kjpp.2023.27.2.131

Figure Lengend Snippet: (A) MiR-665 is predicted as a target of circ_0129047 through CircInteractome. (B) RIP assay was performed using Ago2 antibody in LUAD cells, and the enrichment of circ_0129047 and miR-665 was detected. **p < 0.01 vs . Anti-IgG. (C) Luciferase activity of Circ_0129047 in LUAD cells transfected with miR-665 mimics, which bind to the circ 0129047 sequence. **p < 0.01 vs . mimic-NC. (D) RT-qPCR analysis of miR-665 in LUAD tissues and normal tissues. (E) RT-qPCR analysis of miR-665 in LUAD cells (PC9 and Calu-3) and normal cells (BEAS-2B). **p < 0.01 vs . BEAS-2B. (F) Pearson analysis of miR-665 expression and circ_0129047 expression in LUAD tissues. (G) circ 0129047-overexpressing vectors (OE-circ), OE-NC, miR-665 mimic (mimic), mimic-NC, and OE+mimic were transfected into PC9 and Calu-3 cells. The expression of miR-665 was measured using RT-qPCR 48 h after transfection. **p < 0.01 vs . OE-NC; ## p < 0.01 vs . mimic-NC; && p < 0.01 vs . OE+mimic. Values are presented as mean ± SD. LUAD, lung adenocarcinoma; Ago2, Argonaute 2; WT, wild type; MUT, mutant.

Article Snippet: Human bronchial epithelium (BEAS-2B, cat#: CRL-9609), LUAD cells (A549, cat#: CRM-CCL-185 derived from the lung tissue of a white, 58-year-old male with LUAD; H1975, CRL-5908 derived from the lung tissue of a nonsmoking female with LUAD; and Calu-3, cat#: HTB-55, derived from the lung tissue of a white, 25-year-old male with LUAD) were obtained from the American Type Culture Collection (ATCC), and PC9 (cat#: 90071810, derived from the lung tissue of an undifferentiated patient with LUAD) was obtained from Sigma-Aldrich.

Techniques: Luciferase, Activity Assay, Transfection, Sequencing, Quantitative RT-PCR, Expressing, Mutagenesis

Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Hsa_circ_0129047 sponges miR-665 to attenuate lung adenocarcinoma progression by upregulating protein tyrosine phosphatase receptor type B

doi: 10.4196/kjpp.2023.27.2.131

Figure Lengend Snippet: (A) Predicted miR-665 interactions with PTPRB 3′UTR using miRNA target prediction software based on TargetScan. (B) Luciferase reporter gene assay of PTPRB 3′UTR after miR-665 treatment. **p < 0.01 vs . mimic-NC. (C) PTPRB mRNA expression in LUAD tissues analyzed using RT-qPCR. (D) Pearson correlation analysis was used to analyze the relationship between miR-665 and PTPRB in LUAD tissues. (E) Pearson correlation analysis was used to analyze the relationship between circ_0129047 and PTPRB in LUAD tissues. (F) PTPRB mRNA expression in LUAD cells (PC9 and Calu-3) and normal cells (BEAS-2B) using RT-qPCR. **p < 0.01 vs . BEAS-2B. (G) Western blot analysis determining PTPRB expression in LUAD cells transfected with PTPRB-overexpressing vectors (OE-PTPRB), OE-NC, miR-665 mimic (mimic), mimic-NC, OE-PTPRB+mimic. **p < 0.01 vs . OE-NC; ## p < 0.01 vs . mimic-NC; && p < 0.01 vs . OE-PTPRB+mimic. Values are presented as mean ± SD. LUAD, lung adenocarcinoma; PTPRB, protein tyrosine phosphatase receptor type B; WT, wild type; MUT, mutant.

Article Snippet: Human bronchial epithelium (BEAS-2B, cat#: CRL-9609), LUAD cells (A549, cat#: CRM-CCL-185 derived from the lung tissue of a white, 58-year-old male with LUAD; H1975, CRL-5908 derived from the lung tissue of a nonsmoking female with LUAD; and Calu-3, cat#: HTB-55, derived from the lung tissue of a white, 25-year-old male with LUAD) were obtained from the American Type Culture Collection (ATCC), and PC9 (cat#: 90071810, derived from the lung tissue of an undifferentiated patient with LUAD) was obtained from Sigma-Aldrich.

Techniques: Software, Luciferase, Reporter Gene Assay, Expressing, Quantitative RT-PCR, Western Blot, Transfection, Mutagenesis

Journal: The Korean Journal of Physiology & Pharmacology : Official Journal of the Korean Physiological Society and the Korean Society of Pharmacology

Article Title: Hsa_circ_0129047 sponges miR-665 to attenuate lung adenocarcinoma progression by upregulating protein tyrosine phosphatase receptor type B

doi: 10.4196/kjpp.2023.27.2.131

Figure Lengend Snippet: LUAD cells were transfected with PTPRB-overexpressing vectors (OE-PTPRB), OE-NC, miR-665 mimic (mimic), mimic-NC, and OE-PTPRB+mimic. (A) Cell proliferation was analyzed using the CCK-8 assay. (B) Western blot analysis of anti-Bax and anti-Bcl-2 expression in the transfected cells. (C) Colony formation was determined using colony formation assay (×200). *p < 0.05, **p < 0.01 vs . OE-NC; # p < 0.05, ## p < 0.01 vs . mimic-NC; & p < 0.05, && p < 0.01 vs . OE-PTPRB+mimic. Values are presented as mean ± SD. LUAD, lung adenocarcinoma; OD, optical density; PTPRB, protein tyrosine phosphatase receptor type B.

Article Snippet: Human bronchial epithelium (BEAS-2B, cat#: CRL-9609), LUAD cells (A549, cat#: CRM-CCL-185 derived from the lung tissue of a white, 58-year-old male with LUAD; H1975, CRL-5908 derived from the lung tissue of a nonsmoking female with LUAD; and Calu-3, cat#: HTB-55, derived from the lung tissue of a white, 25-year-old male with LUAD) were obtained from the American Type Culture Collection (ATCC), and PC9 (cat#: 90071810, derived from the lung tissue of an undifferentiated patient with LUAD) was obtained from Sigma-Aldrich.

Techniques: Transfection, CCK-8 Assay, Western Blot, Expressing, Colony Assay