Blocking programmed death-ligand 1 attenuates maturation inhibition of dendritic cells by co-cultured breast cancer cells / 南方医科大学学报

<p><b>OBJECTIVE</b>To study if programmed death-ligand 1 (PL-L1) expression in breast cancer cell activates PD-L1/PD-1 pathway in dendritic cells to inhibit dendritic cell maturation.</p><p><b>METHODS</b>Human monocytes were induced to differentiate into immature dendritic cells using GM-CSF and IL-4, and further to mature dendritic cells using TNF-α. PD-L1-expressing breast cancer cell line MDA-MB-231 was co-cultured in contact with the dendritic cells to observe the effects of the breast cancer cells on the maturation of the dendritic cells. A PD-L1 blocking antibody was applied to the co-culture, and the changes in the inhibitory effect of the MDA-MB-231 cells on dendritic cell maturation was observed. TNF-α-induced dendritic cells were treated with a recombinant human PD-L1 protein to study the effect of PD-L1/PD-1 pathway activation on the maturation of dendritic cells. The expression of PD-L1 in MDA-MB-231 cells and the dendritic cell maturation marker HLA-DR and CD83 were analyzed using flow cytometry.</p><p><b>RESULTS</b>MDA-MB-231 cell line showed PD-L1 positivity on the cell membrane cells at a rate as high as (99.7∓0.15)%. In mature dendritic cells, the positivity rates for HLA-DR and CD83 were (88.8∓6.96)% and (18.36∓3.07)%, respectively, but in the co-culture system, the positivity rates of the dendritic cells were significantly decreased to (42.76∓10.52)% (P<0.01) and (9.93∓2.74)% (P<0.05), respectively, indicating that MDA-MB-231 cells inhibited the maturation of dendritic cells. Following treatment with a PD-L1 antibody isotype control, the percentages of HLA-DR- and CD83-positive cells in the co-culture were (45.17∓10.19)% and (10.15∓2.54)%, which were significantly increased to (63.46∓1.72)% and (16.46∓2.58)% after treatment with PD-L1 antibody, respectively (both P<0.05). Compared with the mature dendritic cell controls, the cells treated with the recombinant human PD-L1 protein exhibited significantly lowered percentages of HLA-DR-positive [from (84.23∓4.18)% to (2.56∓2.39)%, P<0.05] and CD83-positive cells [(87.26∓1.54)% to (60.67∓1.63)%, P<0.05].</p><p><b>CONCLUSION</b>The effect of PD-L1 antibody therapy on triple negative breast cancer can be partially mediated by blocking PD-L1 expression on breast cancer cell membrane, which attenuates the inhibition of dendritic cell maturation in the cancer microenvironment.</p>

Nanog promotes the invasion of breast cancer cells by increasing PKCε expression / 南方医科大学学报

<p><b>OBJECTIVE</b>To study the relationship between Nanog-promoted metastasis of breast cancer and ezrin(T567) phosphorylation, and explore the possible mechanism by which Nanog regulates ezrin(T567) phosphorylation.</p><p><b>METHODS</b>A siRNA construct targeting Nanog was transfected in breast cancer cells to knock down Nanog expression, and the changes in the cell invasion was detected using Transwell assay. The expression levels of Nanog and PKC and the phosphorylation level of ezrin(T567) were detected using Western blotting and immunofluorescent staining; the protein interaction between PKCε and ezrin was assayed by co-immunoprecipitation and Western blotting.</p><p><b>RESULTS</b>Nanog knockdown significantly decreased the expression of PKCε protein, phosphorylation level of ezrin(T567) and the invasion ability of breast cancer cells. PKCε knockdown obviously decreased the phosphorylation level of ezrin(T567) in the cells, and PKCε and ezrin were co-immunoprecipitated.</p><p><b>CONCLUDIONS</b>Nanogcan can upregulate the expression of PKCε to promote the phosphorylation of ezrin(T567), which can be a new mechanism by which Nanog promotes tumor metastasis.</p>

Phosphorylation of ezrin Tyr477 is critical in invasion of breast cancer cells stimulated by precursor of nerve growth factor / 南方医科大学学报

<p><b>OBJECTIVE</b>To investigate the effect of precursor of nerve growth factor (proNGF) in promoting invasion of breast cancer cells and its relation with ezrin expression and phosphorylation of ezrin Thr567 and Tyr477.</p><p><b>METHODS</b>Human breast cancer cell lines MDA-MB-231 and MCF-7 were stimulated by gradient concentrations of proNGF (0, 2.5, 5 and 10 ng/mL) for 16 h, and the invasion of the cells was assessed with Transwell assay. The expression of ezrin and the phosphorylation of ezrin Thr567 and ezrin Tyr477 in the treated cells were examined by Western blotting. MDA-MB-231 cells were transfected with pEnter-His-ezrinY477F (a dominant negative mutant) to study the role of phosphrylation of ezrin Tyr477 in the invasion of breast cancer cell stimulated by proNGF.</p><p><b>RESULTS</b>proNGF significantly promoted MDA-MB-231 and MCF-7 cell invasion in a concentration-dependent manner (P<0.05), and concentration- and time-dependently increased the phosphorylation of ezrin Tyr477 (P<0.05) without affecting the expression of ezrin or the phosphorylation of ezrin Thr567. The specific inhibitor of src, SKI-606, significantly inhibited the phosphorylation of ezrin Tyr477 induced by proNGF. Transfection with pEnter-His- ezrinY477F inhibited proNGF-induced invasion and phosphorylation of ezrin Tyr477 in MDA-MB-231 cells (P<0.05).</p><p><b>CONCLUSION</b>Phosphorylation of ezrin Tyr477 plays a critical role in the invasion of breast cancer cells stimulated by proNGF via proNGF/src/ezrin Tyr477 pathway.</p>

Intervention of xuezhikang on patients of acute coronary syndrome with different levels of blood lipids / 中国中西医结合杂志

<p><b>OBJECTIVE</b>To observe the intervention of Xuezhikang (XZK) on patients of acute coronary syndrome (ACS) with different levels of blood lipids.</p><p><b>METHODS</b>Adopting the double blind randomized controlled method, 105 patients of ACS were divided into two groups. The 53 patients in the treated group, 26 with normal blood lipids (NBL) and 27 with hyperlipemia (HL) were treated with conventional western medicine plus XZK 1.2 g per day for 12 weeks. The 52 patients in the control group, 25 with NBL and 27 with HL, were treated with conventional western medicine alone. Besides, a healthy control group consisted of 40 subjects was set up. The changes of brachial arterial endothelium-dependent diastolic function (FMD) before and after treatment was observed, the changes of blood levels of nitric oxide (NO), endothelin-1 (ET-1), C-reactive protein (CRP) and lipids were also recorded.</p><p><b>RESULTS</b>Before treatment, FMD value and serum NO level were lower and ET-1 and CRP levels in ACS patients were higher than those in the healthy subjects, and a significant correlation existed between CRP, NO and FMD with LDL-C. After treated for 12 weeks, FMD value and serum NO level increased, levels of ET-1 and CRP decreased significantly in the treated group, showing significant difference to those in the control group (P < 0.05, P < 0.01). Serum levels of TC, TG and LDL-C in the treated group lowered significantly. HDL-C level in patients with HL increased significantly while in those with NBL, it showed a trend of increasing but with no statistical significance.</p><p><b>CONCLUSION</b>Applying XZK in ACS patients in early stage, either with NBL or with HL, could improve the endothelial function, antagonize inflammatory response to stabilize the atheromatous plaque.</p>