The Role And Significance Of ASF1B In Ovarian Cancer

Background and ObjectiveOvarian cancer(OC)is one of the three most common gynecological malignancies,and its mortality rate ranks the first among all gynecological malignancies.Because the early symptoms are not obvious,most patients are diagnosed in the late stage,with lack of effective treatment means.The 5-year survival rate of patients is low,and the prognosis is poor.Therefore,there is an urgent need to find new targeted drug therapeutic targets to improve outcomes of patients.ASF1B(Anti-silencing Function1B)is the molecular chaperone protein of histone H3-H4.Studies have found that ASF1 B plays a key role in the occurrence and development of prostate cancer,endometrial cancer,breast cancer,cervical cancer,clear cell carcinoma of the kidney and other malignant tumors,and is closely related to the poor prognosis of patients.However,the role of ASF1 B in ovarian cancer remains unclear.Therefore,the main purpose of this study is to explore the role and significance of ASF1 B in ovarian cancer.MethodsThe expression level of ASF1 B in ovarian cancer and normal ovarian tissue was analyzed based on GEPIA and TCGA data.Collecting 60 primary ovarian cancer tissues of ovarian cancer patients from Jan 2019 to Jan 2020 in Zhengzhou University People’s Hospital,and collected 60 normal ovarian tissue of patients during the same period,using real-time fluorescent quantitative PCR(q RT-PCR)technology to detect ASF1 B expression level in ovarian cancer tissue and normal ovarian tissue,and analyze the ASF1 B the relationship between the expression and clinical pathological parameters.The expression levels of ASF1 B m RNA and protein in different ovarian cancer cell lines(SKOV3,A2780,ES-2 and COC-1 cells)and normal ovarian cell(IOSE-80 cell)were detected by q RT-PCR and Western Blot.SKOV3 and A2780 ovarian cancer cells were transfected with ASF1 B si RNA.The transfection efficiency of ASF1 B si RNA was detected by q RT-PCR and Western Blot to construct a relatively stable ASF1 B knockout cell line.CCK8 assay and cell colony formation assay were used to detect the effect of ASF1 B si RNA on the proliferation and clone formation ability of ovarian cancer cells.Transwell assay was used to detect the effect of ASF1 B si RNA on the invasion ability of ovarian cancer cells,and flow cytometry was used to detect the effect of ASF1 B si RNA on the cell cycle and apoptosis of ovarian cancer cells.The expression levels of apoptosis related proteins and PI3K/Akt/m TOR signaling pathway related proteins in ovarian cancer cells treated with ASF1 B si RNA were detected by Western Blot.Results1.Compared with normal ovarian tissue,the expression level of ASF1 B was significantly increased in ovarian cancer tissue(P<0.001),and the expression of ASF1 B was correlated with FIGO stage,tumor size and lymph node metastasis(P<0.05),but not with age or histological type(P>0.05).2.Compared with normal ovarian cells,the expression level of ASF1 B was significantly increased in various ovarian cancer cell lines(P<0.01);3.In A2780 and SKOV3 cells,ASF1 B si RNA could significantly inhibit the expression of ASF1 B m RNA and protein(P<0.0001);4.In A2780 and SKOV3 cells,the proliferation and clone formation ability of cells in si RNA-ASF1 B group were significantly reduced compared with the control group(P<0.001).The cell invasion ability of si RNA-ASF1 B group was significantly decreased(P<0.001).5.In A2780 and SKOV3 cells showed that compared with the control group,the number of cells in G1 phase was significantly increased in si RNA-ASF1 B group(P<0.001),and the number of cells in G2 and S phase was significantly decreased(P<0.05).Moreover,the number of apoptosis in si RNA-ASF1 B group was significantly increased compared with the control group(P<0.0001).6.In A2780 and SKOV3 cells,compared with the control group,the expressions of pro-apoptotic proteins Bax and lytic caspase-3 were significantly increased in si RNA-ASF1 B group,while the expressions of anti-apoptotic proteins Mcl-1 and Bcl-2 were significantly decreased.7.In A2780 cells and SKOV3 cells,the protein expression levels of p-PI3 K,p-Akt,p-m TOR and PDK1 were significantly decreased in si RNA-ASF1 B group compared with the control group.Conclusion1.The high expression of ASF1 B in ovarian cancer tissues and cells is closely related to FIGO stage,tumor size and lymph node metastasis,and affects the prognosis of patients.2.ASF1 B is involved in the occurrence and development of ovarian cancer by promoting cell proliferation,invasion and inhibiting cell apoptosis;3.ASF1 B is involved in the occurrence and development of ovarian cancer by PI3K/AKT/m TOR signaling pathway;4.ASF1 B is expected to be a new targeted therapeutic target and prognostic indicator for ovarian cancer.