MiR-29b Suppresses The Proliferation And Migration Of Osteosarcoma Cells By Targeting CDK6

[Objective]Literature study showed that miR-29b acts as tumor suppressor of cancer cell in the proliferation and migration processes.CDK6 can promote the cancer cell proliferation and migration.We research whether miR-29b suppresses the proliferation and migration of osteosarcoma cells targeting CDK6.Our results provide theoretical basis for clinical development of osteosarcoma and may open new avenues for osteosarcoma therapies.[Methods]We select patients with osteosarcoma.Selected criteria is without other tumors,non-clinical recurrence,not chemotherapy and radiotherapy.When they accept surgery,we collect some sections of osteosarcoma tissues and adjacent noncancerous tissues.We measure the levels of CDK6 protein of osteosarcoma tissues and adjacent noncancerous tissues via Western blotting.Subsequently,we performed quantitative RT-PCR to measure the levels of CDK6 mRNA of cancerous and noncancerous tissues.After determining the levels of miR-29b of osteosarcoma tissues and adjacent noncancerous tissues,we next investigate whether miR-29b was correlated with CDK6 in osteosarcoma.Three computational algorithms were used in combination to identify miR-29b that can target CDK6.After the recombinant plasmid was transfected into MG-63 cells with pre-miR-29b,we determine whether CDK6 expression of the MG-63 cells were mediated.Furthermore,we introduced point mutations into the corresponding complementary sites in the 3'-UTR of CDK6 to eliminate the predicted miR-29b binding sites.Then,transfected the recombinant plasmid into MG-63 cells with pre-miR-29b,we determine whether CDK6 expression level of the MG-63 cells were affected.Finally,according to contrast principle,we combine pre-miR-29b,pre-miR-control,CDK6 plasmid and control plasmid,and transfect into MG-63 cells using Lipofectamine 2000(Invitrogen).We performed CCK8 assays to determine cell proliferation ability.Transwell assays were performed to determine the cell migration ability.[Results]The upregulation is the expression of CDK6 in osteosarcoma tissues,other than CDK6 mRNA.We investigated that miR-29b is inversely correlated with CDK6 in osteosarcoma.Subsequently,our results demonstrate that miR-29b directly recognizes and binds to the 3'-UTR of the CDK6 mRNA transcript and inhibits CDK6 translation in osteosarcoma cells.MiR-29b suppresses the proliferation and migration of osteosarcoma cells by targeting CDK6[Conclusion]We show that miR-29b inhibite CDK6 expression and consequently inhibite proliferation and migration in cultured osteosarcoma cells.Our results may open new avenues for future osteosarcoma therapies.