Research of cryptochrome gene 1 in human osteosarcoma

Abstract

Objective Aim at investigating the function and mechanism of cryptochrome gene 1 in human osteosarcoma. Methods The lentivirus vector with short hairpin RNA (shRNA) targeting CRY1 was transfected into human osteosarcoma cells (HOS). The knockdown of CRY1 in HOS was confirmed by Western blotting; the proliferation of HOS cells was detected by cell counting kit-8 (CCK-8) assay; the downstream molecules was confirmed by Western blotting; the mRNA expression level of clock gene cryptochrome gene (CRY1, CRY2), period gene (PER1, PER2), BMAL1, CLOCK was tested by quantitative real-time polymerase chain reaction (RT-qPCR). Results After the knockdown of CRY1, the proliferation of HOS was enhanced (0.839±0.020 vs. 0.553±0.019, P<0.01) and the canonical Wnt signaling was found activated (0.729±0.067 vs. 1.119±0.018, P<0.01), as well as the mRNA levels of all clock network key genes but CRY1 were increased. What’s more, proliferation of HOS cell was inhibited in rescue experiment (1.012±0.151 vs. 1.483±0.095 vs. 0.541±0.129, P<0.01). Conclusion CRY1 controls proliferation of human osteosarcoma cells via Wnt signaling pathway and regulates the expression levels of other clock network key genes, including CRY2, PER1, PER2, BMAL1 and CLOCK, which indicates that CRY1 plays an important role in human osteosarcoma progression. Key words: Cryptochrome gene 1; Wnt signaling pathway; Clock gene networks; Osteosarcoma