The Correlation And Clinical Significance Of ING4 And MMP-9 In Breast Carcinoma

ObjectiveTo investigate the expressions of ING4 and MMP-9 in breast cancer tissues,analyzing the correlation between them and the value of clinical application.The expressions of ING4 and MMP-9 can be used to estimate the prognosis of the metastatic potential of breast cancer and to provide certain theoretical basis for guiding clinical individualized treatment.Methods1?The expressions of ING4 and MMP-9 protein were detected by immunohistochemistry in 83 breast cancer tissues.The relationship between these proteins and clinicopathological characteristics was explored,and the correlation between the expressions of ING4 and MMP-9 protein was analyzed.2?The ING4 small interfering RNA(siRNA)was transfected into cultured human breast cancer cell-line MDA-MB-231 by liposome transfection method for knocking down the ING4 express.The cells were divided into three groups: transfection group with ING4-siRNA(experimental group),control-siRNA group(matched group)and blank group with no transfection.3?The expression of ING4 was detected by the real-time PCR and Western blotting,in order to determine whether transfection success and screen ING4 missing cells.4?Transwell invasion and migration assay were used to test the influence of the invasiveness and metastasis on MDA-MB-231 cells silenced ING4.5?Western blotting was conducted to detect the expression of MMP-9 on the there groups respectively.Results1?The positive expression rate of protein ING4 and MMP-9 were 43.4%(36/83)?73.5%(61/83)in tissues of breast cancer.They were significantly associated with the histological grade,lymphnode metastasis and tumorous TNM staging of breast cancer(P<0.05).There was a negative correlation between these proteins.2?The real-time PCR results show that the ING4 m RNA expression quantity in the transfection group with ING4-siRNA is significantly lower than the other two groups among in the three groups of MDA-MB-231 cells,the difference was statistically significant(P<0.01),but the expressions in the remaining two groups has no significant statistical difference(P>0.05).3?The Western blotting results was that the protein expression content in the experimental group is significantly lower than the other two groups among in the three groups(P<0.01),but the amount of ING4 protein expression in the remaining two groups has no significant statistical difference(P>0.1).4?Transwell invasion assay indicate that the number of cells through matrigel was(215.35±8.12)?(118.50±5.03)and(119.72±6.20)in this three groups.Experimental group was higher than any groups(P<0.01),the difference was statistically significant.Transwell migration assay results show that the number of cells was(348.14±10.56)?(238.32±5.12)and(251.23±7.02),the difference was statistically significant(P<0.01),The relationship between the groups is similar with the results of matrigel assay.5?The expression of MMP-9 was detected by Western blotting in this three kinds of cells respectively.The result was that the MMP-9 protein expression quantity was up-regulated after ING4 gene silencing,and the statistical difference between groups was significance(P< 0.01).Conclusions1?The expressions of ING4 and MMP-9 may play important roles in development and progression of breast cancer.There may be mutual synergy effect.Combined detection of these two markers could provide certain reference for early pathological diagnosis ?clinical individualized treatment and prognosis of breast carcinoma.2?The expression of ING4 reduction or deletion not only promote the invasion and metastasis of breast cancer cells,but also increase the expression of MMP-9.