There are three subtypes of undifferentiated human conventional osteosarcoma (HCOS): osteoblastic osteosarcoma (OOS), chondroblastic osteosarcoma (COS), and fibroblastic osteosarcoma (FOS). HCOS also exhibits heterogeneous pathological maldifferentiation in individual patients. Currently, the mechanism regulating HCOS differentiation remains unclear, and therapies are ineffective. Osteopontin (OPN) and osteocalcin (OCN) are markers of osteoblast maturation, and their expression is inhibited in HCOS. A previous study found that PLK2 inhibited TAp73 phosphorylation and consequent anti‐OS function of TAp73 in OS cells with enriched TAp73. TAp73 was also reported to regulate bone cell calcification. Here, OOS was found to have higher TAp73 levels and PLK2 expression than those in COS, which is correlated with HCOS maldifferentiation according to Spearman analysis and affects patient prognosis according to Kaplan‐Meier survival analysis. In the conventional OS cell‐line Saos2 and in patient‐derived xenograft OS (PDX‐OS) cells, increased PLK2 expression owing to abundant TAp73 levels affected OPN and OCN content as measured by RT‐PCR and Western blotting, and alizarin red staining showed that PLK2 affected calcium deposition in OS cells. In addition, PLK2 inhibition in PDX‐OS cells prohibited clone formation, as indicated by a clonogenic assay, and sensitized OS cells to cisplatin (CDDP) (which consequently limited proliferation), as shown by the CCK‐8 assay. In an established PDX animal model with abundant TAp73 levels, PLK2 inhibition or CDDP treatment prevented tumor growth and prolonged median survival. The combined therapeutic effect of PLK2 inhibition with CDDP treatment was better than that of either monotherapy. These results indicate that increased PLK2 levels due to enriched TAp73 affect osteogenic differentiation and maturation and OS prognosis. In conclusion, PLK2 is a potential target for differentiation therapy of OS with enriched TAp73.

Osteosarcoma (OS) universally exhibits heterogeneity and cisplatin (CDDP) resistance.Although the Wee1/CDC2 and NF-κB pathways were reported to show abnormal activation in some tumor cells with CDDP resistance, whether there is any concrete connection is currently unclear. We explored it in human OS cells. Materials and MethodsMultiple OS cell lines were exposed to a Wee1 inhibitor (AZD1775) and CDDP to assess the half-maximal inhibitory concentration values. Western blot, coimmunoprecipitation, confocal immunofluorescence, cell cycle, and CCK-8 assays were performed to explore the connection between the Wee1/CDC2 and NF-κB pathways and their subsequent physiological contribution to CDDP resistance. Finally, CDDP-resistant PDX-OS xenograft models were established to confirm that AZD1775 restores the antitumor effects of CDDP. ResultsA sensitivity hierarchy of OS cells to CDDP and AZD1775 exists. In the highly CDDP-tolerant cell lines, Wee1 and RelA were physically crosslinked, which resulted in increased abundance of phosphorylated CDC2 (Y15) and RelA (S536) and consequent modulation of cell cycle progression, survival and proliferation. Wee1 inhibition restored the effects of CDDP on these processes in CDDP-resistant OS cells. In addition, animal experiments with CDDP-resistant PDX-OS cells showed that AZD1775 combined with CDDP not only restored CDDP efficacy but also amplified AZD1775 in inhibiting tumor growth and prolonged the median survival of the mice. ConclusionSimultaneous enrichment of molecules in the Wee1/CDC2 and NF-κB pathways and their consequent coactivation is a new molecular mechanism of CDDP resistance in OS cells. OS CANCER RESEARCH AND TREATMENT (CRT) 4 Korean Cancer Association This article is protected by copyright. All rights reserved.with this molecular signature may respond well to Wee1 inhibition as an alternative treatment strategy.

Memetic algorithm (MA) is widely applied to optimize routing problems as it provides one way to combine local search with global search. However, the local search in MA needs to be carefully designed according to the problem's characteristics. In this article, we consider a real-world large-scale waste collection problem with multiple depots, multiple disposal facilities, multiple trips, and working time constraints. Vehicles with a limited capacity and working time can start from different depots, collect waste at different sites, and make multiple trips to different disposal facilities to empty the waste and return to its origin. While the existing work considered problems with multiple trips and time constraints, none have tackled problems with multiple depots, multiple disposal facilities, multiple trips, as well as working time constraints. The change from "singledepot" to "multidepot" not only reflects better the situation in real life but also leads to a qualitative different and more complex problem. In this article, we first model this complex problem mathematically. Then, a novel region-focused MA is proposed to tackle this new challenge. Compared to classic MA, this region-focused one is enhanced by two major components: 1) a new heuristic-assisted solution initialization algorithm and 2) a region-focused local search with novel heuristics. Comprehensive computational studies show that our proposed approaches significantly outperform several state-of-the-arts on our real problem of thousands of tasks. The new local search procedure and solution initialization method significantly improve the search ability in combination with global search ability of MA.

No abstract