The Expression Of FASTKD4 In Osteosarcoma And Its Effect On Proliferation And Apoptosis Of Osteosarcoma Cells

Background:Osteosarcoma（OS）is formed by cell mutation（osteoid generation）caused by primitive mesenchymal cells under the action of various reasons.Genome mutation,chromosome modification,tumor suppressor gene disorder and DNA repair defect are the main causes of the occurrence of OSTEosarcoma.The treatment and prognosis of osteosarcoma remains one of the most troubling malignancies for clinicians.Bone cancer,including primary and metastatic bone cancers,remains a challenge that claims millions of lives and affects the quality of life of survivors.Conventional treatment for osteosarcoma includes extensive surgical excision,radiotherapy,and chemotherapy.However,some osteosarcoma cells may remain locally or recrudesce after resection,some are resistant to chemotherapy,some are insensitive to radiotherapy,and the 5-year survival rate is often poor.Therefore,it is urgent to develop specific molecular targeted therapy for patients with poor prognosis of osteosarcoma,so as to further improve the prognosis of patients with osteosarcoma.FASTKD4 is a mitochondrial protein,through influence the expression of mitochondrial RNA that affect the occurrence of tumor development.At present,through the unremitting efforts of a large number of scientific researchers,FASTKD4 is involved in the development and progression of multiple tumors,but whether FASTKD4 has an effect on osteosarcoma has not been studied yet.In order to clarify the role of FASTKD4 in osteosarcoma and further understand the gene and molecular regulatory mechanism related to osteosarcoma,so as to provide a new research direction for improving the survival rate of patients with osteosarcoma.Objective : To explore the expression level of FASTKD4 in osteosarcoma and Paracancerous and its effect on the proliferation and apoptosis rate of osteosarcoma cells,and further explore its molecular mechanism through the PI3K/AKT signaling pathway,the most common abnormal signaling pathway in osteosarcoma.Methods:The levels of FASTKD4 m RNA were detected by q PCR,and protein were detected by immunohistochemistry in osteosarcoma and paracancerous tissues.The levels of FASTKD4 RNA were detected by q PCR in different osteosarcoma cells,and the osteosarcoma cell Saos-2 was used as the research object.The Saos-2 cells were transfected with FASTKD4 small interfering RNA（sh FASTKD4 group）and negative control（sh Ctrl group）.q PCR and Western blot were used to detect the FASTKD4 m RNA and protein in transfected Saos-2 cells.Saos-2 cell hyperplasia and breeding after transfection was detected by multi-parameter high content screening technique and CCK-8 technique.The colony formation ability and apoptosis of Saos-2 cells after transfection were detected by clonal formation assay and flow cytometry.In addition,in order to further explore the molecular mechanism of FASTKD4 occurrence in osteosarcoma,we studied the most common abnormal signaling pathway PI3K/AKT signaling pathway in osteosarcoma,and detected the effect of FASTKD4 knockout on the expression of p-PI3 K,PI3K,p-AKT and AKT in Saos-2 cells by Western blot.Reuslts:The results of q PCR and immunohistochemistry showed the expression of the FASTKD4 in osteosarcoma tissues were higher than those in paracancerous tissues（P<0.05）.The proliferation and colony formation ability of sh FASTKD4 group were suppressed when compared with sh Ctrl group（P<0.05）.However,the percentage of apoptosis in sh FASTKD4 group were higher than those in sh Ctrl group（P<0.05）.Up regulation of FASTKD4 expression in osteosarcoma tissues.Compared with the sh Ctrl group（P<0.05）,knockout FASTKD4 downregulated the expression of p-PI3 K and pAKT proteins in the sh FASTKD4 group（P<0.05）.Conculsion : This study confirmed that FASTKD4 was highly expressed in osteosarcoma tissues,and silencing of FASTKD4 can inhibit proliferation,colony formation of osteosarcoma cell and induce the apoptosis of osteosarcoma cell,We also found that FASTKD4 Knockdown Downregulates PI3K/AKT Signaling pathway in osteosarcoma cells.