| ObjectiveTo study the expression of Valosin-Containing Protein(VCP),NF-κB/P65 in epithelial ovarian carcinoma(EOC)and its relationship with clinicopathological features,and to study the effect of VCP on the migration and proliferation of ovarian cancer cell SKOV-3.To provide experimental basis and new ideas for molecular therapy of ovarian cancer.Methods1.We divided the ovarian tissue samples into four groups,malignant group: 94 cases of EOC tissue samples;Borderline group: 13 cases of ovarian borderline tumor(BOTS)tissue samples;Benign group: 36 cases of ovarian benign tumor tissue samples;Normal group: 8 cases of normal ovarian tissue samples.The expression of VCP and NF-κB/P65 were detected by immunohistochemistry,and the relationship between them and the clinical and pathological parameters(age,FIGO stage,histological grade,pathological subtype,lymph node metastasis,ascites and preoperative CA125)was analyzed.2.The expression of VCP in human ovarian epithelial cell HOSEpiC and epithelial ovarian cancer cell SKOV-3 was detected by immunofluorescence and Western Blot.3.After treated with CB-5083,the migration of SKOV-3 cell at 12 h,24h was analyzed by scratch assay,the proliferation of SKOV-3 cell was analyzed by plate clone formation assay,and the expression of VCP was analyzed by immunofluorescence.The expression of VCP and NF-κB signaling pathway related proteins(NF-κB、 IκB-α、 p-IκBα)were detected by Western Blot.Results1.The positive rate of VCP in EOC,BOTS,benign ovarian tumor and normal ovarian tissue were 73.4%,61.5%,41.7% and 25.0%,respectively,with the difference of them is significant(P < 0.05).Comparing malignant group with other three groups,with the difference of malignant group from benign group and normal group is significant(P < 0.01),with the difference of malignant group and borderline group is no significant(P>0.01).The expression of VCP was correlated with FIGO stage,histological grade and lymph node metastasis(P<0.05),but not with age,pathological subtype,ascites and preoperative CA125(P<0.05).The positive rate of NF-κB/P65 in EOC,BOTS,benign ovarian tumor and normal ovarian tissue were 80.9%,46.2%,41.7% and 37.5%,respectively,with the difference of them is significant(P< 0.05).Comparing malignant group with other three groups,with the difference of the malignant group from the other three groups was significant(P<0.01).The expression of NF-κB/P65 was correlated with FIGO stage,histological grade,ascites and lymph node metastasis(P<0.05),but not with age,pathological subtype and preoperative CA125(P > 0.05).There was a positive correlation between the expression of VCP and NF-κB/P65 in EOC(r=0.38,P<0.05).2.The immunofluorescence intensity of VCP in SKOV-3 cell was significantly higher than that in HOSEpiC cell;The mean relative gray value of VCP was 0.980±0.034 in SKOV-3 cell and that was 0.493 ±0.680 in HOSEpiC cell by Western Blot,with the difference of them is significant(P <0.05).3.After treated with CB-5083,The 12 h migration rate of SKOV-3 cell was 42.14 ±2.15 in the control group and that was 32.22 ±1.96 in the treated group,with the difference of them is significant(P <0.05).The 24 h migration rate was 65.26±2.01 in the control group and that was 40.01±1.28 in the treated group,with the difference of them is significant(P<0.05).The clone formation rate in the control group was 68.80±3.372 and that in the treated group was52.70±3.635,with the difference of them is significant(P<0.05).The mean relative gray value of VCP in the control group was 0.989±0.034 and that in the treated group was 0.728 ±0.057 by Western Blot,with the difference of them is significant(P< 0.05).The mean relative gray value of NF-κB/P65 in the control group was 1.325±0.190 and that in the treated group was 0.575 ±0.104,with the difference of them is significant(P<0.05).The mean relative gray value of IkB-α in the control group was 1.000±0.078 and that in the treated group was 0.910±0.037,with the difference of them is no significant(P<0.05).The mean relative gray value of p-IkBα in the control group was 0.534±0.096 and that in the treated group was 0.867±0.031,with the difference of them is significant(P<0.05).Conclusion1.The positive rate of VCP and NF-κB/P65 in EOC were both increased,and the expression of both were positively correlated.It was concluded that VCP was involved in the carcinogenesis and development of ovarian cancer by activating NF-κB/P65.The combined detection has certain clinical value in evaluating the malignancy and metastasis of ovarian cancer.2.VCP expression in SKOV-3 was higher than that in HOSEpiC and it could promote the proliferation and migration of SKOV-3.It was concluded that VCP could activate the NF-κB pathway by the ubiquitin proteasome pathway. |
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