Journal: Diagnostics

Article Title: Alpha-Enolase (ENO1) Correlates with Invasiveness of Cutaneous Melanoma—An In Vitro and a Clinical Study

doi: 10.3390/diagnostics12020254

Figure Lengend Snippet: The expression level of ENO1 in the cell lysates from primary melanocytes and melanoma cell lines. Representative Western blots showing ENO1 and Akt 1/2/3 expression (for normalization) in protein lysates obtained from the primary human melanocytes (HEM) and indicated melanoma cell lines ( a ). Densitometric ENO1/Akt ratios are shown as mean values ( n = 3 except for HEM, n = 2) ± standard error of the mean (SEM) ( b ). The significance level was set at p = 0.001–0.0001 (***).

Article Snippet: Human epidermal melanocytes, adult (HEMa, 104−05A) and primary human epidermal melanocytes (lightly pigmented) (HEMn-LP, C0025C) were purchased from Cell Applications Inc (San Diego, CA, USA), and Cascade Biologics/Gibco (Carlsbad, CA, USA), respectively.

Techniques: Expressing, Western Blot

Journal: Genome Biology and Evolution

Article Title: Transcriptional Profiling of Human Endogenous Retrovirus Group HERV-K(HML-2) Loci in Melanoma

doi: 10.1093/gbe/evt010

Figure Lengend Snippet: Transcription profiles of HERV-K(HML-2) loci in melanoma. HML-2 gag cDNA sequences were amplified from melanoma cell lines SK-Mel-25, SK-Mel-28, MEWO, and WM3734a, three melanoma RNA samples, two lymph node metastases, and melanocyte cell lines Benno and Oskar. HML-2 loci are indicated on the x axis by chromosomal band and, if available, HGNC approved names. Samples are indicated on the y axis. l. n., lymph node; mel., melanoma. Given on the z axis are relative cloning frequencies as percentages of cDNA sequences that could be assigned unambiguously to particular HML-2 loci, approximately corresponding to transcription levels of those loci in the particular sample. See for further details.

Article Snippet: Melanocyte cell lines Benno and Oskar were cultured in Melanocyte Growth Medium (PromoCell, Heidelberg, Germany).

Techniques: Amplification, Clone Assay

Journal: Genome Biology and Evolution

Article Title: Transcriptional Profiling of Human Endogenous Retrovirus Group HERV-K(HML-2) Loci in Melanoma

doi: 10.1093/gbe/evt010

Figure Lengend Snippet: A HERV-K(HML-2) locus in 2q32.1 transcribed in melanocyte cell line Benno shows typical features of retrotransposition by L1 machinery. ( a ) Depicted in the center-right part of the figure is a dot matrix comparison (window size: 30; minimum score: 50%; jump: 1) of the 2q32.1 locus with the HERV-K(HML-2.HOM) proviral locus (see text) as a reference. Locations of retroviral LTRs, gag , pro , pol, and env gene,s and splice donor (SD) and splice acceptor (SA) sites are indicated for the reference sequence next to the y axis. Subregions in the 2q32.1 locus (chr2 locus), as indicated in the dot matrix comparison, were compared in more detail with a cDNA assigned to this locus (cDNA), a previously reported rec mRNA sequence (Rec) (see text) and the sequence of HERV-K(HML-2.HOM) (HOM). Numbers above the alignment indicate nucleotide positions with respect to the HOM sequence. More detailed sequence comparison shows that the locus represents a retrotransposed HML-2 mRNA that was spliced similar to rec mRNA, with splice donor and acceptor sites resembling the ones in rec mRNA (ii–v). The 2q32.1 locus is flanked by target site duplications (TSDs) and a poly(A)-signal in the 3′-end (i, vi, and vii). The position of the forward primer used for RT-PCR is indicated in (iii). ( b ) Graphical depiction of the HERV-K(HML-2) locus in human chromosome 2q32.1 as provided by UCSC Genome Browser . The retrotransposed HML-2 sequence was detected and annotated by Repeatmasker v.3.2.7, indicated by a black horizontal bar. The same genome region encompassing the HML-2 locus is present in other primate genomes as indicated by greenish or black-colored horizontal boxes. Rhesus and marmoset genomes harbor the same genome region but lack exactly the HML-2 portion as indicated by thin horizontal lines. ( b ) Compiled from annotation tracks provided in hg18 and hg19 ( http://genome.ucsc.edu , last accessed January 31, 2013).

Article Snippet: Melanocyte cell lines Benno and Oskar were cultured in Melanocyte Growth Medium (PromoCell, Heidelberg, Germany).

Techniques: Sequencing, Reverse Transcription Polymerase Chain Reaction

Journal: Genome Biology and Evolution

Article Title: Transcriptional Profiling of Human Endogenous Retrovirus Group HERV-K(HML-2) Loci in Melanoma

doi: 10.1093/gbe/evt010

Figure Lengend Snippet: An HML-2 type II locus in 10q24.2, transcribed in melanocyte cell line Benno, shows a 1,372 bp deletion in env . ( a ) Depicted in the center-right part of the figure is a dot matrix comparison of the ERVK-17 locus in 10q24.2 with HERV-K(HML-2.HOM) and detailed alignments of indicated splice sites of the 10q24.2 genomic sequence (chr10 locus) with two cDNAs assigned to this locus (cDNA1, cDNA2), a rec mRNA sequence (Rec) and HERV-K(HML-2.HOM) sequence (HOM). Numbers above the alignment indicate nucleotide positions in HOM (see also the legend of ). The position of the forward primer used for RT-PCR is indicated in (i). The 10q24.2 locus lacks the intron portion of the rec splice acceptor site (iv). The 5′-end of the missing env portion resembles an alternative splice donor site with the corresponding intron portion missing (iii) approximately 330 bp downstream the regular rec splice donor site (ii). The locus appears to resemble, on the DNA level, another splice variant of HML-2 transcript, and it might represent a spliced HML-2 transcript that was reverse transcribed into a DNA copy in a retroviral fashion. ( b ) The reverse-transcribed locus is present in the human genome but missing in the genomes of non-human primates. See legend of for further details.

Article Snippet: Melanocyte cell lines Benno and Oskar were cultured in Melanocyte Growth Medium (PromoCell, Heidelberg, Germany).

Techniques: Sequencing, Reverse Transcription Polymerase Chain Reaction, Variant Assay

Journal: Genome Biology and Evolution

Article Title: Transcriptional Profiling of Human Endogenous Retrovirus Group HERV-K(HML-2) Loci in Melanoma

doi: 10.1093/gbe/evt010

Figure Lengend Snippet: Quantitation of HERV-K(HML-2) gag transcription by qRT-PCR. ( a ) Relative HML-2 gag transcript levels in melanoma cell lines SK-Mel-25, SK-Mel-28, MEWO, and WM3734a, three melanoma RNA samples, a melanoma lymph node metastasis (LM), and melanocyte cell lines Benno and Oskar, as well as GCT cell lines NCCIT and Tera-1. HML-2 transcript levels were normalized to transcript levels of G6PDH and RPII housekeeping genes. Gag transcript level in melanocyte cell line Benno before UV treatment was taken as reference. Black bars indicate minimum and maximum relative levels of gene expressionas calculated by StepOne software (see Materials and Methods). Note the different scale for NCCIT and Tera-1 cells. ( b ) Relative HML-2 gag transcription in melanoma and melanocyte cell lines before (dark gray bars) and 24 h after irradiation (light gray bars) with 200 mJ/cm 2 UVB.

Article Snippet: Melanocyte cell lines Benno and Oskar were cultured in Melanocyte Growth Medium (PromoCell, Heidelberg, Germany).

Techniques: Quantitation Assay, Quantitative RT-PCR, Software, Irradiation

Journal: Genome Biology and Evolution

Article Title: Transcriptional Profiling of Human Endogenous Retrovirus Group HERV-K(HML-2) Loci in Melanoma

doi: 10.1093/gbe/evt010

Figure Lengend Snippet: Influence of UV irradiation on HERV-K(HML-2) transcription. Melanoma cell lines SK-Mel-25, SK-Mel-28, MEWO, and WM3734a and melanocyte cell lines Benno and Oskar were irradiated with 200 mJ/cm 2 UVB (see text). Given are relative cloning frequencies as percentages of gag- derived cDNA sequences that could be unambiguously assigned to particular HML-2 loci before (dark gray bars) and 24 h after (light gray bars) UVB irradiation of each cell line.

Article Snippet: Melanocyte cell lines Benno and Oskar were cultured in Melanocyte Growth Medium (PromoCell, Heidelberg, Germany).

Techniques: Irradiation, Clone Assay, Derivative Assay

Journal: Cancers

Article Title: Cancer Cell Biomechanical Properties Accompany Tspan8-Dependent Cutaneous Melanoma Invasion.

doi: 10.3390/cancers16040694

Figure Lengend Snippet: Figure 1. Melanoma transformation and progression are associated with stiffness decrease. (a) (Top left panel), overall stiffness measurement of whole human skin reconstructs (HSR) con- taining primary melanocytes (NHM) or melanoma cells. (Top right panel), stiffness measurement of melanoma cells in HSR cryosections. (Bottom panels), imaging of the corresponding HSR sections. (b) Global measurement of skin stiffness of medaka fish mitf::Xmrk/+ (left panel) or mitf::Xmrk/+ p53-/- (right panel) on healthy areas compared to areas with melanoma. Stiffness mea- surements were conducted on the flank or in dorsal areas of medakas. (c) (Upper left panel), optical and mechanical correlative images showing the rigidity of the skin at the interface between healthy area (white area (optical)) and melanoma area (black area (optical)) of the whole mitf::Xmrk/+ medaka. (Bottom left panel), tomographic reconstruction of the area scanned by AFM (projection in xz).

Article Snippet: A melanocyte growth medium kit was used for normal human melanocytes (NHM) (Promocell, Heidelberg, Germany).

Techniques: Transformation Assay, Imaging