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Background Previously, we reported the expression of DLX4 isoforms (BP1 and DLX7) in myeloid leukemia, but the functional role of DLX4 isoforms remains poorly understood. In the work described herein, we further determined the underlying role of DLX4 isoforms in chronic myeloid leukemia (CML) leukemogenesis. Methods The expression and methylation of DLX4 isoforms were detected by real-time quantitative PCR (RT-qPCR) and real-time quantitative methylation-specific PCR (RT-qMSP) in patients with CML. The functional role of DLX4 isoforms was determined in vitro and in vivo. The molecular mechanism of DLX4 isoforms in leukemogenesis was identified based on chromatin immunoprecipitation with high-throughput sequencing (ChIP-Seq)/assay for transposase-accessible chromatin with high-throughput sequencing (ATAC-Seq) and RNA sequencing (RNA-Seq). Results BP1 expression was increased in patients with CML with unmethylated promoter, but DLX7 expression was decreased with hypermethylated promoter. Functionally, overexpression of BP1 increased the proliferation rate of K562 cells with S/G2 promotion, whereas DLX7 overexpression reduced the proliferation rate of K562 cells with G1 arrest. Moreover, K562 cells with BP1 overexpression increased the tumorigenicity in NCG mice, whereas K562 cells with DLX7 overexpression decreased the tumorigenicity. Mechanistically, a total of 91 genes including 79 messenger RNAs (mRNAs) and 12 long noncoding RNAs (lncRNAs) were discovered by ChIP-Seq and RNA-Seq as direct downstream targets of BP1. Among the downstream genes, knockdown of RREB1 and SGMS1-AS1 partially revived the proliferation caused by BP1 overexpression in K562 cells. Similarly, using ATAC-Seq and RNA-Seq, a total of 282 genes including 151 mRNA and 131 lncRNAs were identified as direct downstream targets of DLX7. Knockdown of downstream genes PTPRB and NEAT1 partially revived the proliferation caused by DLX7 overexpression in K562 cells. Finally, we also identified and validated a SGMS1-AS1/miR-181d-5p/SRPK2 competing endogenous RNA (ceRNA) network caused by BP1 overexpression in K562 cells. Conclusions The current findings reveal that DNA methylation-mediated differential expression of DLX4 isoforms BP1 and DLX7 plays opposite functions in leukemogenesis. BP1 plays an oncogenic role in leukemia development, whereas DLX7 acts as a tumor suppressor gene. These results suggest DLX4 as a therapeutic target for antileukemia therapy.

Panax quinquefolium (American ginseng) is a perennial understory herb that has been widely used as a medicinal plant in China and other countries. Autotoxicity has been reported to be one of the major problems hindering the consecutive cultivation of American ginseng. However, the potential autotoxic compounds produced by the root of American ginseng are less well known. Here, we report the isolation and characterization of five groups of autotoxic compounds from aqueous extracts of the fibrous roots of American ginseng. Ether extracts of the water-soluble compounds were further analyzed and separated into seven fractions. Among them, the most autotoxic fraction (Fraction V) was subjected to GC/MS analysis, and 44 compounds were identified. Based on literature information, 14 individual compounds were selected and their autotoxic effects on seedling growth were further tested. The results revealed that, of these 14 compounds, 9 phenolic compounds significantly reduced the growth of seedlings in a concentration-dependent manner, while 5 aliphatic compounds showed modest inhibition at all three concentrations tested. Furthermore, we verified the existence of the autotoxic compounds in the plow layer soil of commercially cultivated American ginseng fields, and the concentration of these compounds as determined by HPLC analysis was in line with the concentration determined to be bioactive. Taken together, our study established a functional link between the compounds produced by American ginseng and their autotoxic effects.