Expulsion Of Micronuclei Containing Amplified Genes Contributes To A Decrease In Double Minute Chromosomes From Malignant Tumor Cells

Double minute chromosomes (DMs) are a hallmark of gene amplification. The relationship between the formation of DMs and the amplification of DM-carried genes remains to be clarified. The human colorectal cancer cell line NCI-H716 and human malignant primitive neuroectodermal tumor cell line SK-PN-DW are known to contain many DMs. To examine the amplification of DM-carried genes in tumor cells, we performed Affymetrix SNP Array 6.0 analyses and verified the regions of amplification in NCI-H716 and SK-PN-DW tumor cells. We identified the amplification regions and the DM-carried genes that were amplified and overexpressed in tumor cells. Using RNA interference, we downregulated seven DM-carried genes, (NDUFB9, MTSS1, NSMCE2, TRIB1, FAM84B, MYC and FGFR2) individually and then investigated the formation of DMs, the amplification of the DM-carried genes, DNA damage and the physiological function of these genes. We found that suppressing the expression of DM-carried genes led to a decrease in the number of DMs and reduced the amplification of the DM-carried genes through the micronuclei expulsion of DMs from the tumor cells. We further detected an increase in the number of H2AX foci in the knockdown cells, which provides a strong link between DNA damage and the loss of DMs. In addition, the loss of DMs and the reduced amplification and expression of the DM-carried genes resulted in a decrease in cell proliferation and invasion ability.What's new? Double-minute chromosomes (DMs) are a hallmark of gene amplification and a major cytogenetic characteristic of malignant tumor cells. The function of DMs and DM-carried genes, however, remains to be clarified. Here, the authors identified amplification regions containing DM-carried genes that were themselves amplified and overexpressed in human malignant tumor cells. Knocking down the DM-carried amplified genes individually, they found that suppression of such oncogenes reduced the number of DMs, the amplification of these DM-carried genes, and cellular function. DNA damage and expulsion of micronuclei containing these DM-carried amplified genes may contribute to the decrease of DMs in tumor cells.