Pleiocarpumlignan A, a new dineolignan from <i>Piper pleiocarpum</i> Chang ex Tseng

The investigation of chemical constituents from the whole plants Piper pleiocarpum Chang ex Tseng resulted in the isolation of one new dineolignan, pleiocarpumlignan A (1), along with one known benzoate derivative, trans-2,3-diacetoxy-1-[(benzoy1oxy)methyl]-cyclohexa-4,6-diene (2), and two known neolignans (3–4) as (±)-trans-dehydrodiisoeugenol (3), (7R,8R,3′S)-△8′-3′,6′-dihydro-3′-methoxy-3,4-methylenedioxy-6′-oxo-8,3′,7,O,4′-lignan (4). Their structures were elucidated through extensive spectroscopic analyses including 1D, 2D NMR, HR-ESI-MS, and by comparison with the literature. All compounds (1–4) were firstly isolated from Piper pleiocarpum Chang ex Tseng. The 13C NMR spectra of 2 were completely assigned for the first time. Cytotoxic activities of these isolated compounds against five human cancer cell lines (including A-549, SMMC-7721, HL-60, MCF-7, and SW-480) were evaluated.</p

CCL22 and TGF-β1 expression in BC.

<p>(A) Negative expression of TGF-β1 in BC (×200). (B) Weak expression of TGF-β1 in BC (×200). (C) Strong expression of TGF-β1 in BC (×200). (D) Negative expression of CCL22 in BC (×200). (E) Weak expression of CCL22 in BC (×200). (F) Strong expression of CCL22 in BC (×200).</p

Prognostic significance of CCL22 Expression in BC.

<p>Kaplan–Meier curves of overall survival (OS) (A) and progression-free survival (PFS) (B) for CCL22 expression in BC to show the association of CCL22 expression with reduced OS (P<0.0001) and PFS (P<0.0001). P values were calculated by the log-rank test.</p

Expression of both CCL22 and Foxp3 ⁺Tregs infiltration in the tumor bed was associated with BC prognosis.

<p>Kaplan-Meier curves are shown for overall survival (OS) (A) and progression-free survival (PFS) (B) was stratified by expressions of two factors to divide the patients into three subsets, CCL22^-Foxp3^low+ group, CCL22^-Foxp3^high+/CCL22⁺Foxp3^low+ group, and CCL22⁺Foxp3^high+ group. The CCL22⁺Foxp3^high+ group was associated with both shorter OS (P=0.001) and PFS (P=0.001) than CCL22^-Foxp3^high+/CCL22⁺Foxp3^low+ group, and CCL22^-Foxp3^low+ group.</p

Syndecan binding protein expression in normal breast and breast cancer tissue.

*<p>, the <i>P</i> value was calculated by Mann–Whitney <i>U</i>-test.</p

Construction of SDCBP silenced breast cancer cells.

<p>(<b>A</b>) Syndecan binding protein (SDCBP) and negative control (NC) short-hairpin RNA (shRNA) expression constructs were transiently transfected into human embryonic kidney 293T cells and the holoproteins in cell lysates were western-blotted for SDCBP shRNA selection. Digits 397, 523, 611 and 987 represent initiation site of four candidate target sequences selected in SDCBP mRNA (NM_001007067) and (*) represent the target sequence we selected as SDCBP shRNA. (<b>B</b>) MDA-MB-231 cells with SDCBP silenced were selected by Western Blot analysis. (*) represented the clone we selected as the MDA-MB-231-SDCBP shRNA cell. (<b>C</b>) BT-549 cells with SDCBP silenced were selected by Western Blot analysis. (*) represented the clone we selected as the BT-549-SDCBP shRNA cell.</p

The effect of SDCBP silence on breast cancer cell cycle.

<p>(<b>A</b>) Flow cytometry were used to analyze the cell cycle. The proportion of cells at G₁, S and G₂/M phase were represented by three independent experiments. (<b>B</b>) The proportions of syndecan binding protein (SDCBP) silenced MDA-MB-231 and BT-549 cells at G₁ were compared with those of their corresponding control shRNA transfected cells at G₁ respectively. **<i>P</i><0.01; ***<i>P</i><0.001 (Student's <i>t</i>-test). (<b>C</b>) The cell cycle regulators p21, p27, phospo-Rb (S780) and cyclin E were western-blotted in above-mentioned cells. β-actin was used as the internal control.</p

Syndecan binding protein expression and pathological features of breast cancers.

*<p>, <i>P</i> values were calculated by Spearman's Rank-Correlation test (n = 160).</p>#<p>, Age: Expressed as median (range), F = 1.105, <i>P</i> = 0.349 (ANOVA test).</p>$<p>, Tumor size: Expressed as mean ± standard deviation, F = 0.153, <i>P</i> = 0.928 (ANOVA test).</p

The relationship between SDCBP expression levels and estrogen treatment in estrogen-responsive MCF-7.

<p>(<b>A</b>) Semi-quantitative reverse transcription-PCR and (<b>B</b>) Western Blot analysis of syndecan binding protein (SDCBP) in MCF-7 cells under different concentrations of 17-β estradiol (E₂) stimulation. β-actin was used as the internal control. (<b>C</b>) Quantitative analysis of SDCBP mRNA level in MCF-7 under steroid hormone deprivation and 10 nM E₂ stimulation by real-time quantitative reverse transcription-PCR. SDCBP mRNA expression levels were normalized against β-actin mRNA expression. Each experiment was repeated three times. **<i>P</i><0.01 (Student's <i>t</i>-test).</p

Syndecan Binding Protein (SDCBP) Is Overexpressed in Estrogen Receptor Negative Breast Cancers, and Is a Potential Promoter for Tumor Proliferation

<div><p>Background</p><p>Syndecan binding protein (SDCBP), an adapter protein containing PDZ domains, contributes to the tumorigenicity and metastasis of many malignant tumors, such as malignant melanoma. Our study aimed in revealing the expression profile of SDCBP in breast cancer (BCa) and its role in tumor cell proliferation, and then exploring its value in the targeted treatment of BCa.</p> <p>Methodology/Principal Findings</p><p>We first evaluated the SDCBP expression by immunohistochemistry in normal breast and BCa tissues. Then we explored the expression profile of SDCBP in different BCa cell lines. By constructing SDCBP-silenced BCa cell clones, we further assessed the effects of SDCBP suppression on tumor cells <i>in vitro</i> by cell culture and <i>in vivo</i> by tumorigenicity. SDCBP expression was detected in 80.6% (n = 160) of BCa tissues, in contrast to its expression in 13% (n = 23) of normal breast tissues (<i>P</i><0.001). Among the tumors, the level of its expression was positively correlated with histological grade and tumor staging while negatively correlated with the estrogen receptor (ER) expression. Higher expression of SDCBP was also noted in ER-negative BCa cell lines. It was also identified that SDCBP expression was down-regulated in a dose-dependent mode by 17-β estradiol in estrogen-responsive MCF-7. Furthermore, SDCBP silence inhibited ER-negative tumor cell growth <i>in vivo</i> and <i>in vitro</i>. Cell cycle studies showed that SDCBP silence increased G1 cell population and resulted in related cell-cycle-regulator changes: up-regulation of p21 and p27 while down-regulation of cyclin E.</p> <p>Conclusion/Significance</p><p>Our results suggested that SDCBP played an important role in tumor growth of ER-negative BCas. In these tumors where the estrogen signaling pathway is not available, SDCBP probably contribute to tumor growth through an alternative signaling pathway by promoting tumor cells passing the G1/S checkpoint into the cell cycle. Suppression of SDCBP expression may have its potential to become a targeted therapy for ER-negative BCas.</p> </div