The Role And Molecular Mechanism Of NUPR1 In The Development Of Ovarian Cancer

BackgroundAmong gynecological malignancies,ovarian cancer(OC)is one of the leading causes of mortality.According to the statistical report from the National Cancer Institute(NCI),there will be 21750 new cases of OC in the United States in 2020,accounting for 1.2%of all new cancer cases and 13940 deaths,accounting for 2.3%of all cancer deaths.OC often occurred in women between 55 and 64 years of age,with a median age of 63 at diagnosis.More than 75%of patients were already in advanced stages(Stage Ⅲ to Ⅳ)at the time of diagnosis.To date,surgery combined with platinum/paclitaxel chemotherapy have been the main treatment for OC.Despite advances in procedures such as cytoreductive surgery and chemotherapy,the five-year overall survival rate for OC patients had changed little since 1980 because the majority of patients were already advanced and chemo-resistant when OC were diagnosed.According to the latest data released by SEER(2010-2016年,https://seer.cancer.gov/statfacts/html/ovary.html),the 5-year survival rate of OC patients in the United States is about 48.6%.Therefore,the searching for potential biomarkers and therapeutic targets is of great significance for the screening and improving the prognosis of OC patients.Iovanna et al.first found overexpression of NUPR1 in acute pancreatitis 20 years ago.Human NUPR1 gene produces two kinds of proteins,NUPR1a,which is composed of 100 amino acids.At present,there is no relevant literature report about NUPR1a.The other is NUPR1b,which is composed of 82 amino acids.It is a small nuclear protein with a molecular weight of 8872.7 Da,later known as P8.Ree later discovered coml(candidate of metatasis-1)through differential display of cDNA from breast cancer metastasis to brain tissue of nude mice,which was a candidate molecule assisting cancer cell metastasis.Coker later found that com1 and P8 were one molecule through gene sequencing,which was later named NUPR1.NUPR1 was associated with many of emergency stimuli.Many factors could lead to the overexpression of NUPR1,such as endothelin and angiotensin,tumor necrosis factor α,demyelinating agents,lipopolysaccharide(LPS),CCl4 and cannabinoid.NUPR1 was also involved in a variety of emergency related functions.Many studies had found that NUPR1 was abnormally expressed in many benign and malignant diseases,such as acute pancreatitis,heart failure,diabetic mesangial cell hypertrophy,breast cancer,brain tumors and brain metastases,thyroid tumors and pituitary tumors.However,two studies based on pancreatic cancer,one confirmed that NUPR1 was overexpressed in most pancreatic cancer specimens,while the other identified NUPR1 as an intracellular growth inhibitor of pancreatic cancer cells.One explanation for these two seemingly contradictory conclusions was that one study was conducted in vivo and the other in vitro,and that different in vivo and in vitro microenvironments lead to different biological functions of NUPR1.NUPR1 had also been shown as an inhibitor in many tumors.For example,studies in prostate cancer had shown that the down-regulation of NUPR1 was negatively correlated with tumor aggressiveness and tumor progression.Therefore,the complex function of NUPR1 might depend on different tumor microenvironments,and the roles of different tissues in different microenvironments might be completely different or even opposite.In conclusion,NUPR1 is a complex molecule with a variety of physiological and biological functions,which has different roles in various benign and malignant tumors.It may be a novel targeted drug gene that intervention with NUPR1 may prevent cancer progression and metastasis.But the role of NUPR1 in OC remains unclear.This study aims to explore the role and molecular mechanism of NUPR1 in the development of OC,and to provide evidence for the targeted treatment of OC with NUPR1.The specific research content includes the following three parts:Part Ⅰ:Expression of NUPR1 protein in OC tissues and its effect on survival of patientsObjective:The expression of NUPR1 protein in OC tissues and its correlation with clinicopathological factors,overall survival and recurrence-free survival in OC patients were investigated.Materials and Methods1.Materials:Information on OC tissue microarray(TMA):The total number of cases in OC tissue microarray were 154 cases,including 13 cases with metastases,4 cases with benign tumors or cancer adjacent tissues,and the remaining 137 cases of primary tumors.6 cases of primary OC tissue were excluded for reasons such as the stripped tissue from the slide.Follow-up were included in the analysis of 131 cases of OC.The clinicopathological data were collected,including the expression of NUPR1 protein,patients age,TNM stage,pathological grade,pathological type,maximum tumor size,Ki67 expression,etc.The first pathological classification group:non serous adenocarcinoma group and serous adenocarcinoma group;the second pathological classification group:type I(low-grade serous carcinoma/endometrioid carcinoma/clear cell carcinoma/mucinous carcinoma,etc.)and type II(high-grade serous carcinoma/sarcoma/undifferentiated carcinoma,etc.);2.Methods:2.1.The expression of NUPR1 protein in OC tissues was detected by immunohistochemistry.These patients were divided into two groups:NUPR1 low-expression group and NUPR1 high-expression group using to the staining intensity and positive rate of immunohistochemistry staining;2.2.Correlation analysis about the expression of NUPR1 protein and clinicopathological features in patients with OC.Chi-square test was used to detect the correlation between the expression level of NUPR1 protein and the clinicopathological characteristics of OC patients.t-test was used to compare the differential expression of NUPR1 protein in primary OC tissues,metastatic tissues,benign tumors or adjacent tissues;2.3.Correlation analysis about the expression of NUPR1 protein and survival rate of OC.Kaplan-Meier method and Log-Rank statistical method were used to analyze the correlation between the expression level of NUPR1 protein and the overall survival rate and recurrence-free survival of OC patients according to the follow-up data and immunohistochemical results.Univariate analysis and COX multivariate regression analysis were used to to analyze prognostic factors related to the overall survival rate and recurrence-free survival in OC patients.Results:1.Expression difference and significance of NUPR1 protein in benign tumor or adjacent tissues of ovary,primary tumor tissues of OC and metastatic tissues of OC:/-test was used to detect the expression of NUPR1 protein in benign tumors or adjacent tissues of ovary,primary tumor tissues of OC,and metastatic tissues of OC.The results showed that the expression of NUPR1 protein in primary tumor tissues and metastatic tissue of OC was higher than that in benign tumor or adjacent tissue,the difference was statistically significant(P values<0.05),suggesting that NUPR1 might be related to the occurrence and development of OC;2.Correlation between the expression level of NUPR1 protein in OC tissues and clinicopathological features of OC patients.The Chi-square test was used to detect the correlation between the expression level of NUPR1 protein and the clinicopathological features in OC patients.The results showed that there was no significant correlation between the expression level of NUPR1 protein of OC tissues and patient age(P=0.167)and pathological grade(P=0.163).It was significantly correlated with TNM stage(P=0.031),pathological type 1(serous and non serous)(P<0.001),pathological type 2(type Ⅰ and Ⅱ)(P=0.016),Ki67 expression(P=0.001)and maximum tumor size(P=0.008),the differences were statistically significant.The results showed that the positive expression of NUPRI protein in TNM stage Ⅲ/Ⅳ,serous adenocarcinoma,type Ⅱ OC and positive expression of Ki67 was higher than respectively to TNM stage Ⅰ/Ⅱ,non serous adenocarcinoma,type Ⅰ ovarian cancer and,negative expression of Ki67;3.Correlation between the expression of NUPR1 protein and the overall survival rate and recurrence-free survival:Kaplan-Meier method and Log-Rank statistical analysis showed that the expression of NUPRI protein in OC tissues was negatively correlated with the overall survival rate and the recurrence-free survival,the differences were statistically significant(both P<0.001).The higher the expression of NUPR1 protein,the lower the overall survival rate,the shorter the recurrence-free survival.The lower the expression of NUPR1 protein,the higher the overall survival rate and the longer the recurrence-free survival,suggesting that high expression of NUPR1 was associated with poor prognosis of OC;4.Analysis of prognostic factors related to the expression level of NUPR1 protein in OC tissues and overall survival and recurrence-free survival of OC patients:Application of univariate and multivariate regression analysis showed that the expression of NUPR1 protein in OC tissues,TNM stage and maximum tumor sizes were independent prognostic factors of overall survival,difference was statistically significant(P values<0.001,<0.001 and=0.026,respectively).OC patients with higher expression of NUPR1 protein,advanced TNM stage(stage III/IV),and larger maximum tumor size(>12cm)had lower overall survival than those with lower expression NUPR1 protein,stage Ⅰ/Ⅱ and maximum tumor size<12cm.TNM stage of OC and maximum tumor sizes were independent prognostic factors for recurrence-free survival,difference was statistically significant(P<0.001 and=0.028,respectively).OC patients with advanced TNM stage(stage Ⅲ/Ⅳ)and larger maximum tumor size(>12cm)had shorter recurrence-free survival than those with stage Ⅰ/Ⅱ and maximum tumor size<12cm.Conclusions:NUPR1 protein is highly expressed in primary and metastatic tissues of OC,which is associated with poor prognosis of OC.The detection of NUPR1 is expected to be a new indicator for diagnosis and prognosis evaluation of OC.Part Ⅱ:Study on the molecular biological functions of NUPR1 in OC cellsObjective:1.The expression of NUPR1 in ovarian surface epithelial(OSE)cells and OC cell lines(A2870 and SKOV3 cells)were tested;2.OC cells with down-expression and over-expression of NUPR1 were constructed to investigate the effects of NUPR1 on the proliferation,migration and invasion of OC cells;3.Tumorigenesis assay was used to verify the effect of NUPR1 on tumor formation of OC cells in vivo.Materials and Methods1.Materials:OSE cells,A2780 cells,SKOV3 cells and 293T cells were all purchased from the Cell Bank of Chinese Academy of Sciences(Shanghai,China).siRNA was entrusted to Shanghai GenePharma Co.,Ltd..Plasmid construction and lentivirus packaging were entrusted to Shanghai Genechem Co.,LTD.,and the transfection part was completed by myself.2.Methods:2.1.To verify the difference in the expression of NUPR1 in OSE cells and OC cells:The expression of NUPR1 in OSE cells,A2780 cells and SKOV3 cells were detected by western blot and reverse transcription-polymerase chain reaction(RT-PCR).To determine the relative expression of NUPR1 in OC cells and OSE cells;2.2.To screen out the siRNA series with the best interference effect:Three siRNA-NUPR1(siRNA-337,siRNA-414,siRNA-850)and a series of siRNA-NC were used to treat A2780 and SKOV3 cells.The interference efficiency of siRNA on NUPR1 was verified by western blot and RT-PCR.The effects of interference on cell proliferation were detected by MTT assays and plane clone formation experiments.The effects of interference on cell migration and invasion were analyzed by transwell assays.The effects of interference on cell cycle and apoptosis were detected by flow cytometry.Combined with the above factors,one siRNA was selected to construct short hairpin RNA(shRNA-NUPR1)with stable and down-expression of NUPR1;2.3.To construct shRNA-NUPR1 and verify the effect of downexpression NUPR1 on OC cells:A2780 cells were treated with shRNA-NUPR1 to establish stable low expression of NUPR1.Western blot assays and RT-PCR assays were used to detect the relative expression of related proteins and mRNA of NUPR1 to verify the transfection efficiency.MTT assays,plane clone formation experiments and EdU assays were used to detect the proliferation of A2780 cells with low expression of NUPR1.Transwell assays were used to detect the migration and invasion ability;2.4.To construct OC cells with overexpression of NUPR1 and verify the effect of overexpression of NUPR1 on OC cells:The overexpression plasmid of NUPR1 was constructed and transfected into A2780 and SKOV3 cells.Western blot assays and RT-PCR assays were used to detect the relative expression of related proteins and mRNA of NUPR1 to verify the transfection efficiency.EdU assays,plane clone formation experiments and MTT assays were used to detect the proliferation of OC cells after overexpression of NUPR1.Transwell assays were used to detect the changes of cell migration and invasion ability;2.5.Tumor formation experiment in nude mice:After stable transfection of A2780 cells with low expression of NUPR1,the tumorigenesis experiment in nude mice was carried out to further verify the effect of NUPR1 silenced OC cells on tumorigenesis in nude mice.Results:1.The expression of NUPR1 in OC cells and OSE cells:Western blot assays showed that the expression of NUPR1 in OC cell lines A2780 and SKOV3 cells was higher than that in OSE cells(P=0.0139 and 0.011,respectively),and the expression of NUPR1 in A2780 cells was higher than that in SKOV3 cells(P=0.0481).RT-PCR tests showed that the expression of NUPR1 mRNA in OC cell lines A2780 and SKOV3 cells was higher than that in OSE cells(P=0.0397 and 0.0372,respectively).And the expression of NUPR1 mRNA in A2780 cells was higher than that in SKOV3 cells,but the difference between the two cells was not statistically significant(P=0.1926);2.Effects of interference of NUPR1 with siRNA on the biological functions in OC cells:Three siRNA-NUPRl(siRNA-337,siRNA-414,siRNA-850)and one siRNA-NC targeting NUPR1 were used to treat A2780 and SKOV3 cells.Western blot assays and RT-PCR assays showed that the expression of NUPR1 in the treatment group was significantly lower than that in control group.Both MTT assays and plate clone formation assays suggested that the interference of NUPR1 could inhibit the proliferation of OC cells.siRNA-NUPR1 could reduce the number of cell clones compared with siRNA-NC in A2780 cells,but only the number of cell clones in siRNA-414 group and siRNA-NC group were significantly different(P=0.0266).NUPR1 knockdown group could inhibit the migration and invasion of OC cells compared with the control group.In A2780 cells,siRNA-414 and siRNA-850 could inhibit the migration of OC cells(P<0.05),while siRNA-337 and siRNA-414 could inhibit the invasion of OC cells(P<0.05).In SKOV3 cells,siRNA-NUPR1 significantly inhibited cell migration and invasion compared with siRNA NC group(P<0.05).Flow cytometry results showed that siRNA-NUPR1 could promote the apoptosis of A2780 and SKOV3 cells,but only siRNA-414 and siRNA-850 could increase the apoptosis of A2780 cells,which showed statistical difference compared with the siRNA-NC group(P<0.01).The proportion of G0/G1 phase cells decreased and the proportion of S phase cells increased in siRNA-NUPR1 group compared with the control group in A2780 cells.Only the change of cell cycle proportion caused by siRNA-414 series was statistically significant compared with the control group(P<0.05);Based on the above results,we selected siRNA-414 series to construct shRNA-NUPR1 with stable and low expression of NUPR1,and applied it in the subsequent experiments;3.Effects of silencing NUPR1 with shRNA on the biological functions of A2780 cells:The results of western blot assays and RT-PCR assays showed that the expression of NUPR1 decreased,the expression of apoptosis related molecules caspase3,caspase9 and Bax increased,the expression of anti-apoptotic molecules Bcl2 and Bcl-xl decreased,and the expression of cell cycle related proteins CDK4 and CDK6 decreased.MTT assays showed that A2780 cells with low expression of NUPR1 had significant inhibition of cell proliferation at 72 h and 96 h(P=0.0009 and 0.0001,respectively).The number of clones in the low expression group were significantly lower than that in the control group(P=0.0301).EdU assays showed that the proliferation of A2780 cells in NUPR1 silencing group was significantly lower than that in control group(P=0.0173).The silencing of NUPR1 could inhibit the migration and invasion of A2780 cells in transwell experiments(P=0.0297 and 0.0156,respectively);4.Effects of overexpression of NUPR1 on the biological functions of OC cells:In OC cells,western blot assays and RT-PCR assays results showed the overexpression of NUPR1 increased the expression of NUPR1,decreased the expression of apoptosis related molecules caspase3,caspase9 and Bax,increased the expression of anti-apoptotic molecules Bcl2 and Bcl-xl,and increased the expression of cell cycle related proteins CDK4 and CDK6.MTT assays,plane clone formation experiments and EdU assays showed that overexpression of NUPR1 could promote the proliferation of OC cells.Transwell assays showed that overexpression of NUPR1 could promote the migration and invasion of OC cells.5.Effect of silencing NUPR1 on tumorigenesis in nude mice in vivo:Tumor formation experiments in nude mice showed that the tumor volume and weight were significantly decreased after NUPR1 was silenced,and the differences were statistically significant compared with the negative control group(P=0.0426 and 0.0443,respectively).Conclusions:NUPR1 is highly expressed in OC cells.Overexpression of NUPR1 can significantly promote the growth of OC cells,and the silencing of NUPR1 can significantly inhibit the growth of OC cells.NUPR1 may be related to the occurrence and development of OC.Part Ⅲ:Molecular mechanism of NUPR1 in proliferation,invasion and migration of OC cellsObjective:The relative expression levels of NUPR1,AKT and p-AKT proteins in A2780 and SKOV3 cells with NUPR1 overexpression treated with AKT pathway inhibitor LY294002 were detected,and the changes of cell proliferation,migration and invasion in each group were compared.Materials and Methods1.Materials:1.1 Cell lines:Human OC cells were the same as the part Ⅱ.It was cultured in RPMI-1640 medium with 10%fetal bovine serum(FBS),1%penicillin and streptomycin double antibody,and incubated aseptically at 37℃ with 5%CO2.1.2.The experimental groups:The experimental groups were divided into four groups:one group was pCMV-NC+DMSO group,i.e.,NC group was added with DMSO;one group was pCMV-NC+LY294002,i.e.,NC group was added with 10μM of AKT inhibitor LY294002;one group was pCMV-NUPR1+DMSO,i.e.,NUPR1 high expression group was added with DMSO;the other group was PCMV-NUPR1+LY294002,in other words,10μM of AKT inhibitor LY294002 was added after high expression of NUPR1.2.Methods:2.1.Changes of related pathway proteins of NUPR1 overexpression in OC cells:Western blot assays were used to detect the relative expression of NUPR1 protein,AKT protein and p-AKT protein in A2780 and SKOV3 cells with NUPR1 overexpression treated with LY294002(pCMV-NC+DMSO group,pCMV-NC+LY294002 group,pCMV-NUPR1+DMSO group and pCMV-NUPR1+LY294002 group);2.2.Changes in proliferation of OC cells:The proliferation and growth of cells treated with LY294002 were detected by MTT assays and plane clone formation experiments:MTT assays and plane clone formation experiments were used to detect the proliferation of the above four groups of cells;2.3.Changes in the migration and invasion ability of OC cells:The migration and invasion ability of OC cells were detected by transwell assays.Results:1.Changes of NUPR1 overexpression related pathway proteins in OC cells:Western blot assays were used to detect the relative expression of NUPR1 protein,AKT protein and p-AKT protein in A2780 and SKOV3 cells treated with LY294002.NUPR1 protein was significantly higher in pCMV-NUPR1+DMSO group and pCMV-NUPR1+LY294002 group than in pCMV-NC+DMSO group in A2780 and SKOV3 cells(all P<0.05).There was no significant difference in NUPR1 protein between pCMV-NUPR1+DMSO group and pCMV-NUPR1+LY294002 group(P>0.05).LY294002 significantly inhibited the expression of p-AKT protein in pCMV-NUPR1+LY294002 group and pCMV-NC+LY294002 group.There was no significant difference in the expression of AKT protein among the groups(all P>0.05);2.Inhibition of AKT pathway can reverse the effect of NUPR1 overexpression on the proliferation of OC cells:MTT assays and plate colony formation assays were used to detect the proliferation and growth of cells treated by LY294002.The results of MTT assays showed that pCMV-NUPR1+DMSO group had higher cell proliferation than pCMV-NUPR1+LY294002 group and pCMV-NC+DMSO group after treatment with LY294002(10 μM)for 72 h in A2780 cells(P=0.,0179 and 0.0443,respectively).pCMV-NC+DMSO group had a higher cell proliferation rate than pCMV-NC+LY294002 group(P<0.05).The pCMV-NUPR1+LY294002 group and the pCMV-NC+DMSO group had similar cell proliferation,and there was no statistical significance between the two groups(P>0.05).pCMV-NUPR1+DMSO group showed higher cell proliferation than pCMV-NUPR1+LY294002 group and pCMV-NC+DMSO group at 96 h after treatment by LY294002(P=0.0008 and<0.0001,respectively).pCMV-NC+LY294002 group could inhibit the proliferation rate of cells compared with pCMV-NC+DMSO group(P<0.05).In SKOV3 cells with NUPR1 overexpression,cell proliferation in the pCMV-NUPR1+DMSO group was significantly faster than that in the pCMV-NUPR1+LY294002 group and the pCMV-NC+DMSO group at 72 h after LY294002 treatment,and the results were statistically significant(all P<0.0001).pCMV-NC+LY294002 group could inhibit cell proliferation rate than pCMV-NC+DMSO group(P=0.0013).However,pCMV-NUPR1+LY294002 group and pCMV-NC+DMSO group had similar cell proliferation rate(P>0.05).Similar results were also observed when SKOV3 cells were treated by LY294002 for 96 h.At 72 h and 96 h after the treatment of SKOV3 cells with LY294002,we compared the ratio of cell proliferation rate of pCMV-NC+LY294002 group and pCMV-NC+DMSO group,and the ratio of cell proliferation rate of pCMV-NUPR1+LY294002 group and pCMV-NUPR1+DMSO group,and found that the ratio of the two groups was statistically significant at 72 h and 96 h(P value was 0.0013 and 0.0006,respectively).These results indicated that LY294002 significantly inhibited cell proliferation in the group with overexpression of NUPR1 compared with the NC group.Inhibition of AKT pathway could reverse the effect of NUPR1 overexpression on the proliferation of OC cells;The pCMV-NUPR1+DMSO group could promote cell proliferation compared with the pCMV-NC+DMSO group in the plane clone formation experiments,and the difference between the two groups was statistically significant(P<0.001),pCMV-NC+LY294002 group could inhibit cell proliferation,the difference between the two groups was statistically significant(P<0.05).pCMV-NUPR1+LY294002 group had higher numbers of cell clone than that in pCMV-NC+DMSO group,there was statistical significance between the two groups(P<0.05).pCMV-NUPR1+DMSO group had higher clon numbers than pCMV-NUPR1+LY294002 group(P<0.05).The ratio of cell clone numbers between pCMV-NC+LY294002 group and pCMV-NC+DMSO group,the ratio of cell clone numbers between pCMV-NUPR1+LY294002 group and pCMV-NUPR+DMSO group were compared,and it showed that the ratio was no statistically significant(P>0.05).In SKOV3 cells with NUPR1 overexpression,compared with the pCMV-NC+DMSO group,the pCMV-NUPR1+DMSO group had a higher numbers of cell clones,and the difference was statistically significant(P=0.0143).pCMV-NUPR1+DMSO group had higher cell clone numbers than pCMV-NUPR1+LY294002 group(P=0.0008).The numbers of cell clones in the pCMV-NC+LY294002 group was lower than that in the pCMV-NC+DMSO group,and the difference was statistically significant(P=0.0426).While there was no significant difference in the numbers of cell clones between the pCMV-NUPR1+LY294002 group and the pCMV-NC+DMSO group(P>0.05).The ratio of cell clone numbers between pCMV-NC+LY294002 group and pCMV-NC+DMSO group and the ratio of cell clone numbers between pCMV-NUPR1+LY294002 group and pCMV-NUPR+DMSO group were compared,and it showed that the ratio of the two groups was statistically significant(P=0.0347).LY294002 significantly inhibited cell proliferation in the group with overexpression of NUPR1 compared with the NC group.This also verified that the inhibition of AKT pathway could reverse the effect of NUPR1 overexpression on the proliferation of OC cells;3.Inhibition of AKT pathway can reverse the effect of NUPR1 overexpression on the migration and invasion of OC cells:Transwell assays were used to detect the migration and invasion of OC cells.A2780 cells in the pCMV-NUPR1+DMSO group showed stronger cell migration ability than the pCMV-NC+DMSO group in the migration experiment(P=0.0001),and the pCMV-NUPR1+DMSO group and the pCMV-NC+DMSO group showed higher cell migration ability compared with the pCMV-NUPR1+LY294002 group(P<0.0001 and=0.0047).pCMV-NC+DMSO group had higher cell migration ability than pCMV-NC+LY294002 group(P=0.0014).The ratio of cell migration ability between pCMV-NC+LY294002 group and pCMV-NC+DMSO group and the ratio of cell migration ability between pCMV-NUPR1+LY294002 group and pCMV-NUPR+DMSO group were compared,and it showed that the ratio of the two groups was statistically significant(P=0.0284).In the invasion experiments,the invasion ability of the pCMV-NUPR1+DMSO group increased compared with the pCMV-NC+DMSO group(P=0.0158),while the pCMV-NUPR1+LY294002 group and the pCMV-NC+DMSO group had similar cell invasion ability(P>0.05).However,pCMV-NC+DMSO group had higher cell invasion ability than pCMV-NC+LY294002 group(P=0.0237).The pCMV-NUPR1+DMSO group had stronger cell invasion ability than the pCMV-NUPR1+LY294002 group(P=0.0103).The ratio of cell invasion ability between pCMV-NC+LY294002 group and pCMV-NC+DMSO group and the ratio of cell invasion ability between pCMV-NUPR1+LY294002 group and pCMV-NUPR+DMSO group were compared,and it was found that the ratio of the two groups was statistically significant(P=0.0359).These results indicated that the A2780 cells with overexpressed with NUPR1 showed strong ability of cell migration and invasion ability,and the ability of cell migration and invasion could be reversed by LY294002.Conclusions:NUPR1 plays a carcinogenic role in promoting the proliferation,migration and invasion of OC cells via the AKT pathway.