Abstract
Purpose: To determine the functional roles of cluster of differentiation 147 (CD147) in glycolysis in melanoma cells.Methods: CD147 expression in melanoma tissue and adjacent normal tissue was determined using quantitative real time polymrase chain reaction (qRT-PCR) and immunohistochemistry. Cell Counting Kit-8 (CCK-8) and colony formation assays were used to evaluate cell viability and colony formation, respectively. The role of CD147 in glycolysis in melanoma cells was investigated by determining glucose uptake, production of lactate, and cellular level of ATP.Results: CD147 was enhanced more in melanoma tissue than that in the adjacent normal tissue (p < 0.001). CD147 overexpression promoted the viability and colony formation of melanoma cells. On the other hand, CD147 silencing decreased the viability and colony formation of melanoma cells. Glucose uptake, production of lactate, and cellular level of ATP were upregulated in melanoma cells by CD147 overexpression and downregulated by shRNA-mediated depletion of CD147. CD147 increased expression of C-X-C motif chemokine ligand 1 (CXCL1) to activate the sex-determining region Y-related high-mobility group box 4 (SOX4) pathway. Knockdown of CXCL1 attenuated the positive regulatory effect of CD147 on SOX4. Besides, overexpression of SOX4 reversed the suppressive effects of CD147 silencing on melanoma cell viability, colony formation, and glycolysis.Conclusion: CD147 contributes to melanoma cell growth via upregulation of SOX-mediated glycolysis, thus providing a therapeutic avenue for the management of melanoma.
 Keywords: Cluster of differentiation 147, CD147, Sex-determining region Y-related high-mobility group box 4, Melanoma, Cell growth, Glycolysis
Highlights
Melanoma is considered as the most aggressive type of skin cancer, and patients with local or distant metastasis of melanoma have a poor prognosis [1]
Results showed that Cluster of differentiation 147 (CD147) was enhanced in melanoma tissue compared to non-tumor tissue (p < 0.001) (Figure 1 A and B)
A significant downregulation of CD147 was verified in SK-MEL-2 cells transfected with shCD147 #1 or #2 compared to that transfected with negative control Short hairpin RNAs (shRNAs) (p < 0.001, Figure 2 A)
Summary
Melanoma is considered as the most aggressive type of skin cancer, and patients with local or distant metastasis of melanoma have a poor prognosis [1]. Little report considering the downstream target involved in CD147-mediated aerobic glycolysis in melanoma cells. This study hypothesized that SOX4 was implicated in CD147-mediated glycolysis of melanoma cells. This study was performed to clarify the functional roles of CD147 in glycolysis and proliferation of melanoma cells, and identify the role of the CD147 was depending on regulating SOX4, providing a therapeutic factor for melanoma treatment. For colony formation assay, A375 or SK-MEL-2 cells were seeded (2 × l02 /well) into 6-well plates, and cultured in DMEM, with the medium replaced with fresh medium every 3 days. Following blocking with 5 % bovine serum albumin in phosphate-buffered saline (PBS) for 1 h, membranes were incubated overnight with specific primary antibodies against CD147. After incubation with the horseradish peroxidase-labeled secondary antibody (1:5000; Abcam), specific protein bands in each membrane were examined.
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