ROCK2 Promotes Osteosarcoma Growth and Glycolysis by Up-Regulating HKII via Phospho-PI3K/AKT Signalling

Abstract

Background: Osteosarcoma (OS) is a malignant bone tumour that exhibits a high mortality. While tumours thrive in a state of malnutrition, the mechanism by which OS cells adapt to metabolic stress through metabolic reprogramming remains unclear. Methods: We analysed the expression of ROCK2 in osteosarcoma tissues by RT-qPCR and Western blot. Cell proliferation were analysed using CCK8, EdU and colony formation assays. The level of cell glycolysis was detected by glucose-6 phosphate, glucose consumption, lactate production and ATP levels. Results: Herein, our study showed that ROCK2 expression in OS tissues was higher than in adjacent tissues. Functional assays have demonstrated that ROCK2 contributes to the growth of OS cells by inducing aerobic glycolysis. The current study revealed that ROCK2 knockdown decreased the levels of mitochondrial hexokinase II (HKII). And also indicated that ROCK2 served as a key enzyme in glycolysis and that it served an important role in tumour growth. A significant positive correlation was identified between the mRNA and protein expressions of ROCK2 and HKII, further demonstrating that ROCK2-induced glycolysis and proliferation was dependent on HKII expression in OS cells. Mechanistically, ROCK2 promotes HKII expression by activating the phospho-PI3K/AKT signalling pathway. Conclusion:Taken together, the results of the current study linked the two drivers of OS growth and aerobic glycolysis and identified a new mechanism of ROCK2 control in OS.

miR‑5590‑3p inhibits the proliferation and metastasis of renal cancer cells by targeting ROCK2 to inhibit proliferation, migration and invasion

“…Multiple studies have detected high expression of ROCK2 in cancer tissues. Deng et al ( 34 ) reported that ROCK2 was upregulated in osteosarcoma tissues compared to those of adjacent ones, and it stimulated the malignant growth of osteosarcoma by upregulating HKII through phosphorylating the PI3K/AKT signaling pathway. Luo et al ( 35 ) discovered that ROCK2 was upregulated in the bladder cancer cell line T24 under hypoxic conditions.…”

Deubiquitinating enzyme USP10 promotes osteosarcoma metastasis and epithelial–mesenchymal transition by stabilizing YAP1

“…The experimental conditions of CHX have been discussed in our previous experimental studies. 26,28 Furthermore, protein degradation kinetic tests revealed a significant extension of the half-life of YAP1 protein in U2-OS and MG63 cells with upregulated USP10 (Figure 8E,F). These results confirm that USP10 affects YAP1 protein degradation.…”

“…qRT-PCR, western blot, and co-IP assays were conducted according to the earlier description. 28 The primers used for qRT-PCR were: USP10: forward: 5-AATAA AGG GAA CTG GTGC-3, reverse: 5-CTATC ATG GGT GTT GACGT-3; YAP1: forward: 5-GCAAC TCC AAC CAG CAG CAACA-3, reverse: 5-CGCAG CCT CTC CTT CTC CATCTG-3; and GAPDH: forward: 5-AGCCT CAA GAT CAT CAG CAATG-3, reverse: 5-CCATC ACG CCA CAG TTTCC-3. The antibodies used for western blotting were: USP10 (Abcam, ab109219, 1:1500 dilution), YAP1 (Abcam, ab52771, 1:1000 dilution), vimentin (Abcam, ab92547, 1:1500 dilution), E-cadherin (Abcam, ab40772, 1:1500 dilution), Ncadherin (Abcam, ab18203, 1:1500 dilution), and GAPDH (Abcam, ab9485, 1:5000 dilution).…”

The roles of glycolysis in osteosarcoma

“…The mRNA and protein expression of glycolytic enzyme HK2 is regulated by ROCK2. The specific mechanism is that ROCK2 promotes the expression of HKII by activating PI3K/AKT signal pathway, and then induces glycolysis and proliferation of OS cells ( Deng et al, 2021 ). In summary, OS cells are maintained and developed through the PI3K/AKT pathway, which mediates glycolysis and proliferation.…”