A Preliminary Interacting Study Of RSK4 And HSP90 In Breast Cancer

Background :Breast cancer has become a threat to women's health in China the main malignant tumors,the incidence rate increased year by year,increase the incidence of breast cancer in China has been ranked in the forefront of the world,and more and more young,for breast cancer treatment along with the gradual replacement of the development of science and technology,put forward from the beginning of last century "the local operation" concept gradually developed into "systemic therapy,molecular targeted therapy is more popular in today's research,found that affect gene proteins in the genesis and development of breast cancer is very necessary and has long-term significance.In the previous study,the interaction of RSK4m2 protein and HSP90aa1 protein in breast cancer cells was found by immunoprecipitation.Objective:The purpose of this study is to find out the relationship between ribosomal S6 protein kinase 4(RSK4)and heat shock protein 90(HSP90)in the presence of interacting proteins in breast cancer,and preliminarily explore the regulation mechanism of RSK4 protein and HSP90 protein in breast cancer.Methods:Construction of RSK4 expression vector in MDA-MB-231 cells,RSK4 and HSP90 in the construction of MDA-MB-231 cell lines of CO-IP protein,MDA-MB-231 cell lines by expression of RSK4 protein in the verification of whether there is an interaction of RSK4 and HSP90 protein by immunoprecipitation;the expression of Western blot and HSP90 MDA-MB-231 in the quantitative detection of RSK4 and MCF-7 cells,the expression of statistical validation of the difference and correlation between rt-qPCR and HSP90 in MDA-MB-231 quantitative detection of RSK4 and MCF-7 cells,statistical validation of difference and correlation respectively;RSK4 protein and HSP90 protein in two with anti planting and fluorescence confocal of dish,the difference and expression of fluorescence by confocal laser microscopy detection of RSK4 and HSP90 in MDA-MB-231 and MCF-7 cells;collected 102 clinical cases of breast cancer patients cancer tissues and paracancerous tissues and its corresponding Pro Pathological information and immunohistochemistry were used to detect the expression of RSK4 and HSP90 in cancer tissues and adjacent tissues.Results:1?The co precipitation protein of RSK4 and HSP90 protein was detected by the immunoprecipitation method,indicating the interaction between the two proteins;2?The results of Western bolt showed that the expression level of RSK4 at MCF-7 protein level was higher than that of MDA-MB-231(t=4.554,P=0.045),and HSP90 expression at MDA-MB-231 protein level was higher than MCF-7(t=6.238,MDA-MB-231).Linear regression showed a negative correlation between the expression of RSK4 and HSP90 at the protein level(r=-0.849,P=0.032);3 ? Immunofluorescence results showed that RSK4 was expressed in cytoplasm in MDA-MB-231 and MCF-7 cells.HSP90 was also expressed in cytoplasm,and rarely expressed in cell membrane and nucleus.Under a laser scanning confocal microscope and RSK4 fluorescence intensity in MCF-7 cells was higher than that of MDA-MB-231,the expression of RSK4 protein in MCF-7 was higher than that in MDA-MB-231 cells,RSK4 protein expression in low-grade malignant tendency in breast cancer cell lines;observation of HSP90 fluorescence intensity in MCF-7 cells was weaker than that of MDA-MB-231 laser scanning confocal microscope,the expression of HSP90 the amount of protein in MCF-7 was lower than that of MDA-MB-231 cells,HSP90 proteins tend to express in highly malignant breast cancer cell lines;4 ? The results of RT-qPCR showed RSK4 mRNA expression in MDA-MB-231 and MCF-7 were 1,1.271 + 0.042,t test showed that RSK4 had significant difference in the expression of two kinds of cells,the expression of MDA-MB-231 cells was lower than that of MCF-7(P=0.001,t=14.41);HSP90 mRNA in MDA-MB-231 and MCF-7 respectively is 1,0.744 + 0.082 T,test showed that there were significant differences in the expression of these two kinds of cells in the HSP90 expression of MDA-MB-231 cells was higher than that of MCF-7(P=0.002,t=6.931).Linear regression showed a negative correlation between the expression of RSK4 and HSP90 at the level of mRNA(P<0.05,r=-0.935);5 ? Immunohistochemical observation showed that RSK4 protein was mainly moderately or weakly positive in breast cancer tissues,while in the adjacent tissues,it was mainly moderately positive,and a few showed strong positive expression.RSK4 in 34 cases(33.7%)cancer tissues,67 cases of adjacent tissues(66.5%)were positive,the positive expression of cancer tissue was lower than that of adjacent tissues(c2=21.018,P<0.05);HSP90 protein showed strong positive or moderate positive expression in breast cancer tissues,adjacent cancer tissues with moderate and weak positive.HSP90 in 75 cases(74.5%)of 30 cases of cancer tissues,paracancerous tissues(29.4%)were positive,the positive expression of cancer tissue was higher than that of adjacent tissues(c2=41.218,P<0.05);chi square test analysis of differentiation and clinicopathologic factors that RSK4 expression level and the organization that RSK4(P=0.000)expression,ER the expression(P=0.038),PR(P=0.002)and HSP90 expression(P=0.004)was correlated with age,tumor size,lymph node metastasis,the expression of HER2 and distant metastasis(P>0.05).Conclusion:(1)low expression of RSK4 in breast cancer and high expression of HSP90;(2)RSK4 and HSP90 have mutual protein interaction in breast cancer;(3)the expression of RSK4 and HSP90 in breast cancer is negatively correlated,but how to regulate and control each other in the molecular mechanism needs further study.