| Objective: To detect the expression of OPCML, the methylationstatus of promoter, and the influence on the multiplatcation, migration andinvasion of human breast cancer MB231cell from the expression of geneOPCML. To investigate the possible mechanism on the down expression ofOPCML and its aberrant methylation during the breast cancer formationprocess.Methods:(1). We detect the protein expression of OPCML in humanbreast cancer tissues, breast cancer adjacent tissues and benign breastlesions by tissue chip technology and immunohistochemistry pv two stepmethod, and analyze the relationship between clinical opothologicalfeatures, general regulation index of breast cancer and the expression ofOPCML. We further investigated OPCML expression in six kinds of humanbreast cell lines （MDA-MB-231, MB468, MCF-7, SK-BR-3, T47D,BT-549） by western blotting.(2). We investigate the possibility of whichgene OPCML as a molecular marker on clinical pathology for breast cancerby analyzing gene OPCML methylation in human breast cancer tissues andbreast cancer adjacent tissues by MSP.(3).We analyze the influence onthe multiplatcation, migration and invasion from gene OPCML by colony-forming unit assays, wound-healing assay and Transwell invasionassay, to understand the biological behavior of gene OPCML on humanbreast cancer cells.Results:(1). PV two step immunoassay showed that the positive rate ofthe expression on OPCML was20.16%(26/129) on human breast cancertissues,90.24%(37/41) on breast cancer adjacent tissues and75.51%(37/49)on benign breast lesions, and there was a statistical significance (P<0.05).The expression of OPCML was positively related with histological grade,pTNM staging and lymph node metastasis of breast carcinoma (P<0.05).And The expression of OPCML has a negative correlation withKi67(r=-0.236)、Her-2（r=-0.262)、EGFR（r=-0.243)(P<0.05). Western blotshowed it was loss expressed on all six human breast cancer cellsMDA-MB-231，MB468，MCF-7，SK-BR-3, T47D，BT-549, but inexpressed on paracarcinoam tissues.(2).140breast cancer cases had amethylation in all172cases, the methylation rate of OPCML was81.40%(140/172), and it was only obviously in the expression of Her-2according the clinical pathological cases (P<0.05).(3). The multiplicationcapacity depressed after OPCML transfect human breast cancer cellMDA-MB-231successfully. The MDA-MB-231cell coloning efficiency ofthe OPCML transfecion group and pcDNA3.1group was (25.87±1.03)%，(47.33±1.27)%,(P<0.05). The migration of MB-231cells in the OPCMLgroup [(28.32±5.15)μm] was inhibited significantly as compared with the empty vector group[(70.74±1.82)μm] and untransfected group[(67.50+3.17)μm],P <0.05), and the invasion of MB-231cells in theOPCML group was suppressed significantly as compared with theuntrensfected group and the empty vector group [(20.25±2.21) vs(40.75±1.71)、(39.50±2.08)]，P<0.05。Conclusion:(1). OPCML was low expressed or loss expressed inhunan breast cancer tissues and human breast cancer cell strains, it’sexpression was obviously related with hisologic grade, lymphnodetransplant and pathological grade, and it’s expression was negativelycorrelated with the expression of Her-2、Ki-67and EGFR;(2). There washypermethylation of OPCML in human breast cancer tissues. There was apositive correlation with the expression of OPCML and Her-2, but therewas no significant correlation between the age, grade, stage and hormonereceptor status.(3). The expression of OPCML can inhibite the proliferation,migration and invation of human breast cancer MDA-MB-231cell. OPCMLwas a potential tumor suppressor gene. |
PDF Full Text Request