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. 2021 Dec 22;23(1):72. doi: 10.3390/ijms23010072

Figure 1.

Figure 1

Subtraction hybridization identifies MDA-7/IL-24. Schematic of DISH (differentiation induction subtraction hybridization), an approach for identifying and cloning genes associated with the induction of terminal differentiation in human melanoma cells. Treatment of HO-1 human melanoma cells with a combination of IFN-β + mezerein results in a rapid and irreversible loss of proliferation, extinction of tumorigenic potential, and terminal differentiation [12,13,14,15,16]. The DISH approach was developed to identify and clone genes associated with and causative of the physiologic changes associated with terminal differentiation. mRNAs were isolated from actively proliferating and IFN-β + mezerein (2000 units/mL + 10 ng/mL)-treated HO-1 cells that span the first 24 h of treatment, and were converted into cDNAs. Subtraction hybridization was then done between differentiation inducer-treated and control-proliferating cancer cells resulting in the production of a subtracted cDNA library enriched for melanoma differentiation associated (mda) genes. Probing of clones isolated from this cDNA library permitted cloning of mda genes involved in critical cellular processes, some are listed here [12,13,14,15,16]. Reprinted by permission from Fisher, 2005 [17].