The rearrangement of pre-existing genes has long been thought of as the major mode of new gene generation. Recently, de novo gene birth from non-genic DNA was found to be an alternative mechanism to generate novel protein-coding genes. However, its functional role in human disease remains largely unknown. Here we show that NCYM, a cis-antisense gene of the MYCN oncogene, initially thought to be a large non-coding RNA, encodes a de novo evolved protein regulating the pathogenesis of human cancers, particularly neuroblastoma. The NCYM gene is evolutionally conserved only in the taxonomic group containing humans and chimpanzees. In primary human neuroblastomas, NCYM is 100% co-amplified and co-expressed with MYCN, and NCYM mRNA expression is associated with poor clinical outcome. MYCN directly transactivates both NCYM and MYCN mRNA, whereas NCYM stabilizes MYCN protein by inhibiting the activity of GSK3β, a kinase that promotes MYCN degradation. In contrast to MYCN transgenic mice, neuroblastomas in MYCN/NCYM double transgenic mice were frequently accompanied by distant metastases, behavior reminiscent of human neuroblastomas with MYCN amplification. The NCYM protein also interacts with GSK3β, thereby stabilizing the MYCN protein in the tumors of the MYCN/NCYM double transgenic mice. Thus, these results suggest that GSK3β inhibition by NCYM stabilizes the MYCN protein both in vitro and in vivo. Furthermore, the survival of MYCN transgenic mice bearing neuroblastoma was improved by treatment with NVP-BEZ235, a dual PI3K/mTOR inhibitor shown to destabilize MYCN via GSK3β activation. In contrast, tumors caused in MYCN/NCYM double transgenic mice showed chemo-resistance to the drug. Collectively, our results show that NCYM is the first de novo evolved protein known to act as an oncopromoting factor in human cancer, and suggest that de novo evolved proteins may functionally characterize human disease.

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Human anaplastic lymphoma kinase (ALK) has been identified as an oncogene that is mutated or amplified in NBLs. To obtain a better understanding of the molecular events associated with ALK in the pathogenesis of NBL, it is necessary to clarify how ALK gene contributes to NBL progression. In the present study, we found that ALK expression was significantly high in NBL clinical samples with amplified MYCN (n = 126, P < 0.01) and in developing tumors of MYCN-transgenic mice. Indeed, promoter analysis revealed that ALK is a direct transcriptional target of MYCN. Overexpression and knockdown of ALK demonstrated its function in cell proliferation, migration and invasion. Moreover, treatment with an ALK inhibitor, TAE-684, efficiently suppressed such biological effects in MYCN amplified cells and tumor growth of the xenograft in mice. Our present findings explore the fundamental understanding of ALK in order to develop novel therapeutic tools by targeting ALK for aggressive NBL treatment.

The aim of this study is to evaluate the impact of quick response (QR) codes on customer satisfaction and purchase intention in the online shopping context on the basis of perceived flow. This study also showed the impact of QR codes on perceived flow, followed by perceived flow on purchase intention and customer satisfaction, and finally the combination impact of perceived flow and customer satisfaction on purchase intention. Stimulus-Organism-Response theoretical model was used in this research. Data were collected by online questionnaires from 420 valid respondents who purchased a product online via QR code. The covariance-based structural equation modeling approach was used to analyze the structural model and measurements. This study showed that QR codes have a great impact on purchase intention and customer satisfaction. The findings of this article confirmed that QR codes influence perceived flow, combined effect of them, in turn, influenced online shoppers' satisfaction and, finally, purchase intention. These findings are practically significant for both marketers and customers. Marketers should use QR codes as an embedded advertising tool with particular URLs that will lead to more profitability for shopping agency. Besides customers can also learn their behavior regarding QR codes in the online shopping context. This research has great real implications in the field of Internet shopping, online marketing, retailing, marketing promotion, and consumer behavior. Embedded QR codes with advertising are very helpful for advertisers to share and influence customer satisfaction and purchase intention. Sharing information via QR codes will influence customers to engage and share information with other customers in the online community, which will help to increase the volume of sales rapidly. To the best of our knowledge there is no research to evaluate the real impact of QR codes on purchase intention and customer satisfaction on the basis of perceived flow.