Journal: Cells

Article Title: Gelsolin Contributes to the Motility of A375 Melanoma Cells and This Activity Is Mediated by the Fibrous Extracellular Matrix Protein Profile

doi: 10.3390/cells10081848

Figure Lengend Snippet: Phenotypic heterogeneity in terms of GSN level in melanoma cell lines. ( A ) The A375 cells growing on a non-coated surface were fixed and stained to detect GSN and cell nuclei. The white arrow indicates a cell with a high GSN level, the pink arrow a cell with a medium GSN level, and the yellow arrow a cell with a low GSN level. ( B ) Estimation of the distribution of cells with high, medium or low GSN levels within the A375 cells population. The percentage for every group is presented on a given section of the bar (10 images and at least 159 cells were analyzed). ( C ) F-actin subcellular localization in cells with different GSN levels. The cells seeded onto a non-coated surface were fixed and stained with fluorescently labeled phalloidin, anti-GSN antibodies, and Hoechst 33342. Scale bar: 10 μm. ( D ) Immunostaining of other melanoma cell lines, i.e., SK-MEL-28 and WM1341D with antibodies directed against GSN and Hoechst 33342 to detect GSN and cell nuclei. The white arrows indicate cells with high GSN levels. Scale bar: 25 μm.

Article Snippet: WM1341D and SK-MEL-28 cells were from Rockland Immunochemicals Inc. (Pottstown, PA, USA) and CLS GmbH (Eppelheim, Germany), respectively.

Techniques: Staining, Labeling, Immunostaining

Journal: International Journal of Oncology

Article Title: Linc00961 inhibits the proliferation and invasion of skin melanoma by targeting the miR-367/PTEN axis

doi: 10.3892/ijo.2019.4848

Figure Lengend Snippet: Effects of Linc00961 on the proliferation and apoptosis of A375 and SK-MEL-28 cells. Effects of Linc00961 on the proliferation of (A) A375 and (B) SK-MEL-28 cells, as determined using a Cell Counting Kit-8 assay. (C) Effects of Linc00961 on the apoptosis of A375 and SK-MEL-28 cells, detected via flow cytometry. (D) Statistical analysis of the early apoptotic rates of A375 and SK-MEL-28 cells infected with Lv-control and Lv-Linc00961. (E) Tumor growth curve of A375 cells transfected with Lv-control and Lv-Linc00961. (F) Digital pictures of the resected tumors of animals injected with A375 cells infected with Lv-control (n=3) and Lv-Linc00961 (n=3), captured after 35 days. The maximum diameters of Lv-control and Lv-Linc00961 were 1.718 and 0.737 cm, respectively. Scale bar=1 cm. Data from three experiments are presented as the mean ± SD. * P<0.05 vs. Lv-control. Linc00961, long intergenic noncoding RNA 00961; Lv, lentivirus; PI, propidium iodide.

Article Snippet: Human A375 (cat. no. GCC-ME0001CS), SK-MEL-28 (cat. no GCC-ME0003CS) and A2058 (cat. no. GCC-ME0004CS) SM cell lines, and human epidermal HEMa-LP melanocytes (cat. no. GCC-NC0027CS) were all purchased from Shanghai GeneChem Co., Ltd., where they were characterized by mycoplasma detection, isozyme detection and DNA fingerprinting.

Techniques: Cell Counting, Flow Cytometry, Infection, Transfection, Injection

Journal: International Journal of Oncology

Article Title: Linc00961 inhibits the proliferation and invasion of skin melanoma by targeting the miR-367/PTEN axis

doi: 10.3892/ijo.2019.4848

Figure Lengend Snippet: Effects of Linc00961 on the migration and invasion of SM cells. (A) Effects of Linc00961 on the migration of A375 and SK-MEL-28 cells, as determined by a Transwell assay. (B) Statistical analysis of the numbers of migratory A375 and SK-MEL-228 cells. (C) Effects of Linc00961 on cell invasion of A375 and SK-MEL-28 cells, as determined by a Matrigel-coated Transwell assay. (D) Statistical analysis of the numbers of invasive A375 and SK-MEL-228 cells. Scale bar=20 µ m. Data from three experiments are presented as the mean ± SD. * P<0.05 vs. Lv-control. LINC00961, long intergenic noncoding RNA 00961; Lv, lentivirus.

Article Snippet: Human A375 (cat. no. GCC-ME0001CS), SK-MEL-28 (cat. no GCC-ME0003CS) and A2058 (cat. no. GCC-ME0004CS) SM cell lines, and human epidermal HEMa-LP melanocytes (cat. no. GCC-NC0027CS) were all purchased from Shanghai GeneChem Co., Ltd., where they were characterized by mycoplasma detection, isozyme detection and DNA fingerprinting.

Techniques: Migration, Transwell Assay

Journal: International Journal of Oncology

Article Title: Linc00961 inhibits the proliferation and invasion of skin melanoma by targeting the miR-367/PTEN axis

doi: 10.3892/ijo.2019.4848

Figure Lengend Snippet: Linc00961 expression is inversely correlated with miR-367 expression in SM. Effects of Linc00961 on the expression of miRNAs that possessed putative binding sites for Linc00961 in (A) A375 and (B) SK-MEL-28 cells. (C) Expression levels of miR-367 in SM and BN tissues. (D) Association between miR-367 and Linc00961 in SM tissues. (E) Expression levels of miR-367 in SM cells and human epidermal melanocytes. (F) WT and MUT targeting regions of Linc00961 for miR-367. Data from three experiments are presented as the mean ± SD. * P<0.05 vs. control (Lv-control, BN, HEMa-LP). Linc00961, long intergenic noncoding RNA 00961; miR, microRNA; SM, skin melanoma; BN, benign nevi; WT, wild-type; MUT, mutant.

Article Snippet: Human A375 (cat. no. GCC-ME0001CS), SK-MEL-28 (cat. no GCC-ME0003CS) and A2058 (cat. no. GCC-ME0004CS) SM cell lines, and human epidermal HEMa-LP melanocytes (cat. no. GCC-NC0027CS) were all purchased from Shanghai GeneChem Co., Ltd., where they were characterized by mycoplasma detection, isozyme detection and DNA fingerprinting.

Techniques: Expressing, Binding Assay, Mutagenesis

Journal: International Journal of Oncology

Article Title: Linc00961 inhibits the proliferation and invasion of skin melanoma by targeting the miR-367/PTEN axis

doi: 10.3892/ijo.2019.4848

Figure Lengend Snippet: Linc00961 directly sponges miR-367 in SM cells. (A) Effects of Linc00961 on the expression of miR-367 in SM cells. Relative luciferase activity in (B) A375 and (C) SK-MEL-28 cells co-transfected with Linc00961-WT or LINC00961-MUT luciferase plasmid and miR-367 NC or mimics. (D) Detection of LINC00961 in RNA pulled down by Bio-Linc00961, Bio-Linc00961-MUT or Bio-NC probes. Detection of miR-367 in RNA pulled down by Bio-Linc00961, Bio-Linc00961-MUT or Bio-NC probes in (E) A375 and (F) SK-MEL-28 cells. Data from three experiments are presented as the mean ± SD. * P<0.05 vs. control (Lv-control, miR-367 NC, Bio-NC-probe). Linc00961, long intergenic noncoding RNA 00961; miR, microRNA; SM, skin melanoma; NC, negative control; WT, wild-type; MUT, mutant; Bio, biotinylated.

Article Snippet: Human A375 (cat. no. GCC-ME0001CS), SK-MEL-28 (cat. no GCC-ME0003CS) and A2058 (cat. no. GCC-ME0004CS) SM cell lines, and human epidermal HEMa-LP melanocytes (cat. no. GCC-NC0027CS) were all purchased from Shanghai GeneChem Co., Ltd., where they were characterized by mycoplasma detection, isozyme detection and DNA fingerprinting.

Techniques: Expressing, Luciferase, Activity Assay, Transfection, Plasmid Preparation, Negative Control, Mutagenesis

Journal: International Journal of Oncology

Article Title: Linc00961 inhibits the proliferation and invasion of skin melanoma by targeting the miR-367/PTEN axis

doi: 10.3892/ijo.2019.4848

Figure Lengend Snippet: Upregulation of miR-367 reverses the effects of Linc00961 on the proliferation and apoptosis of skin melanoma cells. (A) Expression of miR-367 in A375 and SK-MEL-28 cells transfected with Lv-control + miR-367 NC, Lv-Linc00961 + miR-367 NC or Lv-Linc00961 + miR-367 mimics, as determined via reverse transcription-quantitative PCR. Proliferation of (B) A375 and (C) SK-MEL-28 cells transfected with Lv-control + miR-367 NC, Lv-Linc00961 + miR-367 NC or Lv-Linc00961 + miR-367 mimics, as determined by a Cell Counting Kit-8 assay. (D) Apoptosis of A375 and SK-MEL-28 cells transfected with Lv-control + miR-367 NC, Lv-Linc00961 + miR-367 NC or Lv-Linc00961 + miR-367 mimics, as determined via flow cytometry. (E) Statistical analysis of early apoptotic rates of A375 and SK-MEL-228 cells transfected with Lv-control + miR-367 NC, Lv-Linc00961 + miR-367 NC or Lv-Linc00961 + miR-367 mimics. Data from three experiments are presented as the mean ± SD. * P<0.05 vs. Lv-control + miR-367 NC. Linc00961, long intergenic noncoding RNA 00961; Lv, lentivirus; miR, microRNA; NC, negative control; PI, propidium iodide.

Article Snippet: Human A375 (cat. no. GCC-ME0001CS), SK-MEL-28 (cat. no GCC-ME0003CS) and A2058 (cat. no. GCC-ME0004CS) SM cell lines, and human epidermal HEMa-LP melanocytes (cat. no. GCC-NC0027CS) were all purchased from Shanghai GeneChem Co., Ltd., where they were characterized by mycoplasma detection, isozyme detection and DNA fingerprinting.

Techniques: Expressing, Transfection, Real-time Polymerase Chain Reaction, Cell Counting, Flow Cytometry, Negative Control

Journal: International Journal of Oncology

Article Title: Linc00961 inhibits the proliferation and invasion of skin melanoma by targeting the miR-367/PTEN axis

doi: 10.3892/ijo.2019.4848

Figure Lengend Snippet: Upregulation of miR-367 reverses the effects of Linc00961 on the migration and invasion of SM cells. (A) Numbers of migratory A375 and SK-MEL-28 cells transfected with Lv-control + miR-367 NC, Lv-Linc00961 + miR-367 NC or Lv-Linc00961 + miR-367 mimics, determined by a Transwell assay. (B) Statistical analysis of the numbers of migratory A375 and SK-MEL-228 cells. (C) Numbers of invasive A375 and SK-MEL-28 cells transfected with Lv-control + miR-367 NC, Lv-Linc00961 + miR-367 NC or Lv-Linc00961 + miR-367 mimics, as determined by a Transwell assay. (D) Statistical analysis of the numbers of invasive A375 and SK-MEL-228 cells. Scale bar=20 µ m. Data from three experiments are presented as the mean ± SD. * P<0.05 vs. Lv-control + miR-367 NC. Linc00961, long intergenic noncoding RNA 00961; Lv, lentivirus; miR, microRNA; NC, negative control.

Article Snippet: Human A375 (cat. no. GCC-ME0001CS), SK-MEL-28 (cat. no GCC-ME0003CS) and A2058 (cat. no. GCC-ME0004CS) SM cell lines, and human epidermal HEMa-LP melanocytes (cat. no. GCC-NC0027CS) were all purchased from Shanghai GeneChem Co., Ltd., where they were characterized by mycoplasma detection, isozyme detection and DNA fingerprinting.

Techniques: Migration, Transfection, Transwell Assay, Negative Control

Journal: International Journal of Oncology

Article Title: Linc00961 inhibits the proliferation and invasion of skin melanoma by targeting the miR-367/PTEN axis

doi: 10.3892/ijo.2019.4848

Figure Lengend Snippet: Silencing PTEN reverses the inhibitory effects of Linc00961 on the proliferation and invasion of SM cells. (A) mRNA and (B) protein expression of PTEN in A375 and SK MEL-28 cells transfected with Lv-control + miR-367 NC, Lv-Linc00961 + miR-367 NC or Lv-Linc00961 + miR-367 mimics. Proliferation of (C) A375 and (D) SK-MEL-28 cells transfected with Lv-control + siRNA NC, Lv-Linc00961 + siRNA NC or Lv-Linc00961 + PTEN siRNA, as determined by a Cell Counting Kit-8 assay. (E) Apoptosis of A375 and SK-MEL-28 cells transfected with Lv-control + siRNA NC, Lv-Linc00961 + siRNA NC or Lv-Linc00961 + PTEN siRNA, as determined via flow cytometry. Numbers of (F) migratory and (G) invasive A375 and SK-MEL-28 cells transfected with Lv-control + siRNA NC, Lv-Linc00961 + siRNA NC, or Lv-Linc00961 + PTEN siRNA, as determined via Transwell assays. Data from three experiments are presented as mean ± SD. * P<0.05 vs. Lv-control + miR-367 NC or siRNA NC. Linc00961, long intergenic noncoding RNA 00961; PTEN, phosphate and tension homology deleted on chromosome 10; Lv, lentivirus; miR, microRNA; siRNA, small interfering RNA; NC, negative control.

Article Snippet: Human A375 (cat. no. GCC-ME0001CS), SK-MEL-28 (cat. no GCC-ME0003CS) and A2058 (cat. no. GCC-ME0004CS) SM cell lines, and human epidermal HEMa-LP melanocytes (cat. no. GCC-NC0027CS) were all purchased from Shanghai GeneChem Co., Ltd., where they were characterized by mycoplasma detection, isozyme detection and DNA fingerprinting.

Techniques: Expressing, Transfection, Cell Counting, Flow Cytometry, Small Interfering RNA, Negative Control