The Effect Study On Metastasis And Invasion In Human Breast Cancer Cells By TLR2Signaling Pathway

Breast cancer is one of the most common malignant tumors in female cancer patients. Themetastatic of cancer cells is the leading cause of death in patients with breast cancer, and it isalso one of the biggest problems in breast cancer treatment. So it has been paid close attention tostudy prevention and clinical treatment of breast cancer.It has vital significance to doing researchfor the metastatic mechanism of breast cancer in the treatment of breast cancer.Toll-like receptors (Toll-like receptors, TLRs) is one of the most important andevolutionary conservative pattern recognition receptors (PPRs). They mainly express on humanimmune cells and epithelial cells. By identifying pathogen associated molecular patterns(PAMPs), they play a key role in the process of congenital immune reactions in vivo. Recentstudies have found that TLRs express on many human malignant cancer cells, including breastcancer and it play an important function in the process of development and metastasis of tumor.The prior research in our laboratory found that toll-like receptor2(Toll-like receptor2,TLR2) express differently in human breast cancer cells MDA-MB-231(high metastaticbreast cancer cell line) and MCF-7(low metastatic breast cancer cell line). TLR2expresshigher in MDA-MB-231cells than in MCF–7cells, its amount of expression differ about1500times. Whether it is the different expression of TLR2that lead to the differences betweenthe two kinds of cells in ability to metastasize? What is its mechanism? To solve these problems,our research use the two kinds of human breast cancer cell line MDA-MB-231and MCF-7and LTA as activator of TLR2to specifically activate TLR2signaling pathways to research forthe impact of TLR2signaling pathway in breast cancer cell metastasis ability and discusses themechanism of TLR2signaling pathways in the process of tumor metastasis, and providetheoretical basis for the study of the mechanism of the occurrence and metastasis in malignanttumor.In this study, by the MTT method, cells wound healing experiment, Transwell matrixattacks and soft AGAR cloning experiments we detect thevitro proliferation ability, transferattack ability and the anchor growth ability of the two kinds of human breast cancer cells after LTA stimulus, finding that the LTA stimulation can improve cell proliferation ability of thebreast cancer cell.The cells proliferation ability after LTA stimulation is about2times morecompared with the control group which did not receive the stimulus in MDA-MB-231cell.The enhancement extent is higher than MCF-7.And the enhancement extent of cell attackability and anchoring growth ability after LTA stimulus in MDA-MB-231are all significantlyhigher than the MCF-7cells. Then we blocked TLR2expression with TLR2specific antibodyin the two kinds of cells,we found that cell transfer ability and anchor growth ability of thetwo kinds of cells decreased to varying degrees, the reduction of capabilities of the MDA-MB-231were significantly obvious than MCF-7. Thus we think the different expression ofTLR2is the key factor causing different transfer ability of the two kinds of breast cancer cells.Using ELISA experiment technology after we detected the expression of tumor metastasisrelated factors VEGF, IL-6and TGF-beta1in the two kinds of cells after LTA stimulationand the secretion changes of these factors after TLR2blocking. We found that in MDA-MB-231cell after LTA stimulation the secretion of VEGF, IL-6and TGF-beta1weresignificantly improved, and it is1.5times,1.3times and1.2times in increase range higherthan MCF-7cell. And after sealing process with TLR2antibody the tumor metastasis relatedfactors of the two kinds of cells also reduced, the reduction of capabilities of the MDA-MB-231were significantly obvious than MCF-7. So we conclude that the TLR2signalingpathways may be induced by LTA activation to induce the secretion of tumor metastasis relatedfactors to influence the changes of human breast cancer cell invasion ability.Finally, using JNK inhibitor SP600125to inhibit the activity of the key kinase JNK inTLR2signaling pathways, we detected that when the JNK kinase activity of human breastcancer cell are restricted, cell invasion and the anchor-independent grow abilities after LTAstimulation are inhibited in different degree, and the degree of inhibition in MDA-MB-231cell lines is higher than MCF–7. It demonstrates that JNK plays an important role in TLR2signaling pathway mediates the invasion mechanism in human breast cancer cell.Taken together, through doing the research of the role of TLR2signaling pathways play inthe process of human breast cancer cell metastasis ability and the possible molecularmechanisms. It provides a new clue for doing research on the transfer and infiltration in human breast cancer cells and a theoretical basis for clinical treatment for breast cancer.