| Background Breast cancer is the most common malignancy in women worldwide, and it’s a serious threat to female’s health and quality of life. Metastatic breast cancer is the leading cause of death, and its occurrence and development of a multi-gene, the result of the combined effects of multiple steps. Although most patients with early breast cancer after individualized and comprehensive treatment, can get better disease-free survival and overall survival, however, some patients are still due to chemotherapy treatment resistant, hormone receptor-negative and the lack of specific therapeutic target for other reasons of short-term recurrence and distant metastasis, and ultimately leads to death. Therefore, to explore the proliferation of breast cancer, metastasis mechanism is particularly important.High mobility group protein B1(high mobility group box 1, HMGB1) is a non-histone chromatin-binding protein, as a pro-inflammatory cytokines and growth factors involved in a variety of tumors, invasion and metastasis of malignant process. HMGB1 can facilitate the migration of immune cells through activation of nuclear transcription factor(NF-κB),Nuclear factor-κB(NF-κB) is an anti-apoptotic factor in their family, the vast majority of NF-κB p65, tumor invasion and metastasis of the biological behavior of NF-κB activation of matrix metalloproteinases(MMP) related. Therefore, The focus of this study is to explore the HMGB1 and NF-κB and MMP-9 in tumor proliferation and metastasis.Objective To investigate the effect of HMGB1 role in the malignant behavior of breast cancer cell proliferation, migration and invasion, such as on and explore possible mechanisms to provide a new direction for the future of breast cancer developing new therapeutic strategies.Methods 1.Using MTT method to detect different concentrations of exogenous HMGB1(100-800 ng / ml) on the breast cancer cell line MDA-MB-231 proliferation.2. Using Transwell trial to evaluate exogenous HMGB1(400ng / ml) on the breast cancer cell line MDA-MB-231 migration and invasion effect.3. Using RT-PCR method to detect the gene expression level of P65 and MMP-9 in breast cancer cell line MDA-MB-231 after different concentrations of exogenous HMGB1(100-800ng/ml).4. Using Western-blot method to detect the expression at the protein level of P65 and MMP-9 in breast cancer cell line MDA-MB-231 after different concentrations of exogenous HMGB1(100-800ng/ml).Result 1.MTT results showed that: Compared with the control group, the different dose groups HMGB1 stimulate breast cancer cell line MDA-MB-231 36 h later, all of the MDA-MB-231 cells were stimulated to proliferate, the difference was statistically significant(P <0.05); when the concentration increases from 100 ng / ml to 400 ng / ml, there are also differences between different concentrations(P <0.05), whereas no significant difference between 400 ng / ml group and 800 ng / ml group(P> 0.05).2. Cell migration test results showed that: Compared with the control group, the 400 ng / ml HMGB1 role transference number MDA-MB-231 cells after 36 h increased 2.17 times(± 0.21)(P <0.05). Cell invasion test showed: Compared with control group, 400 ng / ml HMGB1 effect MDA-MB-231 cells 36 h after passing through the increased number of Matrigel 2.58 times(± 0.13)(P <0.05).3. PT-PCR test results showed that::(1) relative to the control group, no different concentrations of HMGB1 m RNA expression after treatment MDA-MB-231 cells p65 significant difference(P> 0.05), and between the various treatment groups p65 m RNA expression of no significant difference(P> 0.05).(2) different concentrations of HMGB1 treated MDA-MB-231 cell m RNA expression of MMP-9 were higher(P <0.05), and between the various treatment groups ratio of MMP-9 m RNA expression of pairwise comparisons were statistical differences significance(P <0.05).4. Western-blot test results showed that:(1) HMGB1 protein p65 treatment group was significantly higher expression levels(P <0.05), and in comparisons between the treatment groups, and no significant difference between p65 expression 100 ng / ml group and 200 ng / ml group(P> 0.05); p65 expression between 400 ng / ml group and 800 ng / ml group had no significant difference(P> 0.05). Protein expression levels(2) HMGB1-treated group was significantly higher than that of MMP-9(P <0.05), and MMP-9 between each treatment group differences in the ratio of pairwise comparisons were statistically significant(P <0.05).Conclusion: exogenous HMGB1 could promote breast cancer cell proliferation, migration and invasion, and it May be related to NF- κ B signaling pathway... |
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