Biomarker for identification of melanoma tumor cells
A technology for melanoma and cells, which is applied in antineoplastic drugs, biological testing, biomaterial analysis, etc., and can solve the problems of accurately identifying the longitudinal growth and metastasis process of melanoma
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Embodiment 1
[0067] Example 1 Observation of melanoma endothelial cell communication in vitro
[0068] The interaction of tumor cells with adjacent endothelial cells is critical for tumor cell survival and metastasis. Melanoma is notorious for its ability to metastasize at a relatively early stage of development. Its aggressive behavior relies at least in part on the interaction between tumor cells and their surrounding stromal tissue. Neuropilin-2 (NRP2), a cell surface involved in angiogenesis and axon guidance, using a prolonged heterotypic cell co-culture system as a model for tumor-stromal cell interactions that occur during metastasis in vivo receptor, a gene thought to be highly upregulated in melanoma cells during their interaction with endothelial cells.
[0069] Three different in vitro melanoma-endothelial cell two-dimensional co-culture systems were used to identify molecular determinants of melanoma-endothelial cell communication ( Figure 1A ). These systems are designed t...
Embodiment 2
[0081] Example 2 Different gene expression during melanoma-endothelial cell communication
[0082] Among co-culture methods, random mixing methods are particularly advantageous for large-scale analysis of changes in the genetic composition of co-cultured cells due to the similar relative numbers and extensive interfaces between heterogeneous cells, which make most The cells may be involved in cell-to-cell, endothelial and melanoma cell communication. Therefore, gene expression profiles of sorted cells were determined using a random mixing approach followed by sorting of heterotypic cells based on cell type-specific fluorescent markers.
[0083] To define the molecular pathways governing melanoma-HUVEC communication, the formation of gene expression profiles of GFP-1205Lu and RFP-HUVEC alone or in mixed cultures for 48 h was performed ( Figure 2A ). Microarray analysis was performed as previously described [5] . Microsoft EXCEL generated a spreadsheet including the express...
Embodiment 3
[0091] Example 3. NRP2 is upregulated during melanoma-endothelial communication and expressed in metastatic melanoma cells
[0092] NRPs are transmembrane glycoproteins that regulate the nervous and vascular systems[17-19] development. They function as co-receptors interacting with the vascular endothelial growth factor (VEGF) receptor and plexin, and bind two receptors known to have different functions: type 3 semaphorin, related to the axis synapse guidance; and members of the VEGF family known to promote angiogenesis. Recent studies have shown that blocking the function of NRP2 can inhibit the metastasis of tumor cells by affecting the metastasis of lymphatic endothelial cells and the formation of tumor-associated lymphocytes. [20] . NRPs are also involved in tumorigenesis as they are expressed in various cancers [17，21-27] superior. Furthermore, NRP2 has recently been shown to regulate processes critical to angiogenesis and melanoma metastasis in vivo [30] , but its r...
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