The Role Of Glucose-regulated Protein 78(GRP78)Affect Invasion And Metastasis In Ovarian Cancer Via MEK/Erk Signaling Pathway

Objective:1.To observe the effect of GRP78 on invasion and metastasis of ovarian cancer cells.2.To investigate the role of MEK/Erk signaling pathway in GRP78 regulation of invasion and metastasis of ovarian cancer cells.Methods:1.Immunohistochemistry and Western blotting were used to detect the positive expression of GRP78 in 60 ovarian serous carcinoma and 40 ovarian serous cystadenoma tissues.SPSS was used to analyze the relationship between the clinical indicators of ovarian serous carcinoma and GRP78 expression.2.Western blot assay was used to determine the expression levels of GRP78,p-MEK,MEK,p-Erk and Erk in HO-8910 cells and HO-8910PMcells.3.Transwell assay was used to detect the invasion and migration abilities of HO-8910 cells and HO-8910PMcells.4.The pcDNA/GRP78 and GRP78-shRNA plasmid was constructed and was respectively transfected into HO-8910 cells and HO-8910PM cells via Lipofectamine-TM2000 mediation,the expression levels of GRP78,p-MEK,MEK,p-Erk and Erk in HO-8910 cells and HO-8910PM cells was determined with Western blot assay,the invasion and migration abilities of HO-8910 cells and HO-8910PM cells was detected by transwell assay.5.After inhibited the MEK/Erk signaling pathway used by PD98095 in HO-8910PM cells,the expression levels of p-MEK,MEK,p-Erk,Erk were determined with western blot assay,the invasion and migration abilities of HO-8910PM cells was detected by transwell assay.Results:1.GRP78 expression was significantly increased in ovarian serous carcinoma tissuesImmunohistochemical results showed that the positive expression rate of GRP78 in ovarian serous carcinoma tissues was 90%,significantly higher than that in ovarian serous cystadenoma tissues(25%)(P<0.01).Western blot results showed that the expression level of GRP78 in ovarian serous carcinoma(88.75%)was significantly higher than that in ovarian serous cystadenoma(11.25%)(P<0.05).Patients with ovarian serous cancer with high expression of GRP78 had higher clinicopathological grading,higher staging,higher level of CA125,and a higher proportion of patients with lymph node metastasis,pelvic metastasis(P<0.05).Western blot results in vitro showed that GRP78 expression in HO-8910PMcells with high metastatic potential was significantly higher than that in HO-8910 cells(with low metastatic potential)(53.05%)(P<0.01).2.HO-8910PM cells undergone significantly invasion and migrationTranswell assay showed that compared with HO-8910 cells,the abilities of invasion and migration of HO-8910PM cells was significantly increased 69.73%,76.08%(P<0.05).3.GRP78 affects the abilities of invasion and metastasis in ovarian cancer cellsAfter overexpressed GRP78 in HO-8910 cells,transwell assay showed that the abilities of invasion and migration of HO-8910 cells was significantly increased 68.32%,72.53%(P<0.05).After silenced GRP78 in HO-8910PM cells,transwell assay showed that the abilities of invasion and migration of HO-8910PM cells was significantly decreased 62.45%,70.18%(P<0.05).4.GRP78 affects MEK/Erk signaling pathway in ovarian cancer cells.After silenced GRP78 in HO-8910PM cells,western blot assay showed that the expression of p-MEK,p-Erk was significantly decreased 40.33%,50.14%(P<0.05).5.Regulating of MEK/Erk signaling pathway can affect invasion and metastasis in ovarian cancer cellsWestern blot assay showed that compared with HO-8910 cells,the expression of p-MEK,p-Erk in HO-8910PM cells was significantly increased 51.32%,38.46%(P<0.05).After used PD98095 to block MEK/Erk signaling pathway in HO-8910PM cells,transwell assay showed that compared with control group,the abilities of invasion and migration of HO-8910PM cells was significantly decreased in PD98095 group 20.16%,34.28%(P<0.05).Conclusion:1.GRP78 affects the abilities of invasion and migration in ovarian cancer cells.2.GRP78 affects the abilities of invasion and migration in ovarian cancer cells via the MEK/Erk signaling pathway.