High-Efficiency Stable Gene Transduction in Primary Human Melanocytes Using a Lentiviral Expression System

Dunlap, Shariff; Yu, Xiaobing; Cheng, Linzhao; Civin, Curt I.; Alani, Rhoda M.

doi:10.1046/j.0022-202x.2004.22214.x

Cited by 12 publications

(14 citation statements)

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“…H23 cells were transduced with Id1 using a bicistronic lentiviral expression system also encoding GFP and a GFP lentiviral vector as a control as previously reported [24, 28]. H23 cells were efficiently transduced by the lentiviral vectors as assessed by GFP expression (Figure 2(a)) and Id1 expression was efficiently induced as demonstrated by Western blotting for Id1 (Figure 2(b)).…”

Section: Resultsmentioning

confidence: 65%

Id1 Promotes Tumor Cell Migration in Nonsmall Cell Lung Cancers

Bhattacharya

Kowalski

Larson

et al. 2010

Journal of Oncology

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Id1, which belongs to the Id family of helix-loop-helix transcription factors has been most associated with tumor progression and metastatsis; however, its significance in lung cancers has not been extensively explored. Here we seek to evaluate the expression of Id1 in a pilot study of nonsmall-cell lung cancers (NSCLCs) and determine its diagnostic and functional significance in these tumors. Paired normal and malignant lung tissues as well as a panel of NSCLC primary tumors and cell lines were evaluated for Id1 expression using Western blotting and quantitative RT-PCR. Functional assays were performed to evaluate the role of Id1 in tumor cell growth, migration and progression. We find Id1 expression is upregulated in squamous cell carcinoma when compared to adenocarcinoma of the lung and that expression of Id1 versus the normal control is variable in NSCLCs. We also note that Id1 expression in NSCLC cells is largely growth factor dependant and constitutive expression of Id1 in NSCLC cells significantly increases tumor cell migration without affecting cell proliferation. We conclude that Id1, as a mediator of tumor cell migration, may be an indicator of aggressive potential in nonsmall-cell lung cancers.

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Section: Resultsmentioning

confidence: 65%

Id1 Promotes Tumor Cell Migration in Nonsmall Cell Lung Cancers

Bhattacharya

Kowalski

Larson

et al. 2010

Journal of Oncology

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“…Additional tumor cell lines were obtained as gifts from various laboratories at Johns Hopkins University and are described in the Supplementary Materials and Methods. GFP and RFP labeling of cells was performed as previously described (18). All cell lines were passaged less than 6 months in culture after which time cells were re-initiated in culture from frozen stocks.…”

Section: Methodsmentioning

confidence: 99%

Integration of Genotypic and Phenotypic Screening Reveals Molecular Mediators of Melanoma–Stromal Interaction

et al. 2011

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Tumor-endothelium interactions are critical for tumor survival and metastasis. Melanomas can rapidly metastasize early in tumor progression, but the dependence of this aggressive behavior on tumor-stromal interaction is poorly understood. To probe the mechanisms involved, we developed a heterotypic coculture methodology, allowing simultaneous tracking of genomic and phenotypic changes in interacting tumor and endothelial cells in vitro. We found a dramatic rearrangement of endothelial cell networks into patterns reminiscent of vascular beds, even on plastic and glass. Multiple genes were upregulated in the process, many coding for cell surface and secreted proteins, including Neuropilin-2 (NRP2). A critical role of NRP2 in coordinated cell patterning and growth was confirmed using the coculture system. We conclude that NRP2 represents an important mediator of melanoma-endothelial interactions. Furthermore, the described methodology represents a powerful yet simple system to elucidate heterotypic intercellular interactions mediating diverse physiological and pathological processes. Cancer Res; 71(7); 2433-44. Ó2011 AACR.

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“…Once considered as an uncommon disease, the incidence of malignant melanoma has doubled within the last 10 years in the US (Balch et al, 2001). It is estimated that an American developing melanoma during his/her lifetime leaped from 1 in 1500 in 1960, to 1 in 68 in 2000, and is projected to increase to 1 in 50 by the year 2010 (Rigel, 2002;Dunlap et al, 2004). Primary melanoma usually progresses from the nonmetastatic radial growth phase to the vertical growth phase, in which the malignant cells invade the dermis and develop the ability to metastasize (Elder, 1999).…”

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confidence: 99%

Nuclear ING2 expression is reduced in human cutaneous melanomas

Lü¹,

Dai²,

Martinka

et al. 2006

Br J Cancer

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Cutaneous malignant melanoma is a severe and sometimes life-threatening cancer. The molecular mechanism of melanomagenesis is incompletely understood. Deregulation of apoptosis is probably one of the key factors contributing to the progression of melanoma. The inhibitor of growth (ING) family proteins are candidate tumour suppressors which play important roles in apoptosis. Downregulated expression of ING proteins have been reported in several tumour types, including the loss of nuclear expression of p33ING1b in melanoma. As ING2 exhibits 58.9% homology with p33ING1b, we hypothesized that the aberrant expression of ING2 may be involved in melanomagenesis. Here, we used tissue microarray technology and immunohistochemistry to examine ING2 expression in human nevi and melanoma biopsies. Our data showed that nuclear ING2 expression was significantly reduced in radial growth phase (RGP), vertical growth phase (VGP), and metastatic melanomas compared with dysplastic nevi (Po0.05). Our data also revealed that nuclear ING2 expression was not associated with patient's gender, age or tumour thickness, ulceration, American Joint Committee on Cancer (AJCC) stage, tumour subtype, location and 5-year survival (P40.05). Taken together, our results suggest that nuclear ING2 expression is significantly reduced in human melanomas and that reduced ING2 may be an important molecular event in the initiation of melanoma development.

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High-Efficiency Stable Gene Transduction in Primary Human Melanocytes Using a Lentiviral Expression System

Abstract: We are grateful to M. Devlin for technical assistance and L. Meszler in the Cell Imaging Core Facility at The Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins for her expertise and assistance in fluorescence imaging of cells.

Cited by 12 publications

References 10 publications

Id1 Promotes Tumor Cell Migration in Nonsmall Cell Lung Cancers

Id1 Promotes Tumor Cell Migration in Nonsmall Cell Lung Cancers

Integration of Genotypic and Phenotypic Screening Reveals Molecular Mediators of Melanoma–Stromal Interaction

Nuclear ING2 expression is reduced in human cutaneous melanomas

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