Three new glycosides from the stems of Eurya chinensis R. Br.

Two new phenolic glycosides (1 and 2), one known analogue (3), along with a new diterpene glucoside (4) were obtained from ethanolic extract of the stems of Eurya chinensis R. Br. The structures of these isolated compounds were identified by extensive analysis of HRESIMS and NMR spectroscopic data. The cytotoxicities of these compounds were evaluated on MCF-7, A549, HepG2, CaCo2 and 5-8 F cell lines by MTT method, but no obvious activities were observed.

Two new withanolides from the whole plants of Physalis peruviana.

Two new withaphysalin-type withanolides (18-O-ethylwithaphysalin R and 5-O-ethylphysaminimin C, 1 and 2), along with twelve known withanolides (3-14), were purified and identified from Physalis peruviana L. The chemical structures of these new isolates were elucidated through analyzing spectroscopic and HRESIMS data. All the obtained metabolites were appraised for their potential antiproliferative activity against the human breast cancer cell line MCF-7. Compound 7 was discovered to exhibit potent activity with an IC50 value of 3.51 µM and compounds 2, 6 and 14 showed weak cytotoxic effect.

Peruranolides A-D, four new withanolides with potential antibacterial and cytotoxic activity from Physalis peruviana L.

BACKGROUND: Many drugs for anti-tumour have been developed, nevertheless, seeking new anticancer drug is the focus of ongoing investigation. Withanolides have been reported to possess potent antiproliferative activity. Literature findings revealed that a diversity of withanolides were obtained from Physalis peruviana, however, the antitumor activity of these bioactive compounds is still unclear. METHODS: The EtOAc fraction of P. peruviana were decolorized on Middle Chromatogram Isolated (MCI) Gel column, repeatedly subjected to column chromatography (CC) over sephadex LH-20, preparative High Performance Liquid Chromatography (HPLC) and silica gel to afford compounds. Their chemical structures of the new isolates were elucidated through analyzing spectroscopic and HRESIMS data. All these obtained metabolites were appraised for their potential antiproliferative activity against the human breast cancer cell line MCF-7 by MTT assay, and in vitro antibacterial activity of the isolated compounds (1-7) were evaluated against E. coli, B. cereus and S. aureus. Results: Four new withanolides, including one withaphysalin-type withanolide (peruranolide A, 1), two 13,14-seco-withaphysalins (peruranolides B-C, 2-3), as well as one normal withanolide (peruranolide D, 4), were purified and separated from P. peruviana L.. Compound 5 was discovered to exhibit potent cytotoxic effect with an IC50 value of 3.51 µM. In vitro antibacterial activities, compounds 1-7 had no obvious inhibitory activity against E. coli, but had moderate inhibitory activities against B. cereus and S. aureus. CONCLUSIONS: Our findings might offer valuable clues for the utilization of withanolides as lead compounds for antineoplastic or antibacterial drug development.

Two new 7,20-epoxy-ent-kaurane diterpenoids from the aerial parts of Isodon serra.

Two new compounds (1 and 2), belonging to C-20 oxygenated ent-kauranes-type diterpenoids, were identified from the aerial parts of Isodon serra. Their structures were elucidated by extensive analysis of HRESI-MS and NMR spectroscopic data. Both these two compounds possess a common 7,20-epoxy-ent-kauranes skeleton with a hydroxyl group rarely occurring at C-13. Compounds 1 and 2 were evaluated for their cytotoxic activity against Hela-60 and HepG2 as well as the antibacterial activity against Staphylococcus aureus, Bacillus cereus and Escherichia coli.

Rosanortriterpenes A-B, Two Promising Agents from Rosa laevigata var. leiocapus, Alleviate Inflammatory Responses and Liver Fibrosis in In Vitro Cell Models.

Rosanortriterpenes A-B (RTA and RTB), two nortriterpenoids, are characteristic constituents in the fruits of Rosa laevigata var. leiocapus. However, pharmacological studies on these compounds are still scarce. In the present study, we aim to investigate the anti-inflammatory mechanisms associated with the effects of RTA-B in RAW264.7 macrophages and LO2 cells by detecting cell viabilities, nitric oxide (NO) production, tumour necrosis factor-alpha (TNF-α), and interleukin-6 (IL-6) production. Simultaneously, the anti-inflammatory action mechanisms of these two compounds were illustrated through western blot assay. Besides, the antihepatic fibrosis activities of these compounds have also been explored. The results demonstrated that RTA and RTB inhibited the production of NO, TNF-α, and IL-6 and suppressed liver fibrosis. RTA and RTB treatment also greatly inhibited the activation of NF-kappaB (NF-κB) pathway. Our study confirmed the promising anti-inflammatory and anti-liver fibrosis actions of RTA-B, suggesting that they might be developed as alternative and promising drugs for the treatment of hepatic inflammatory and fibrotic diseases.

Two new ent-kaurane diterpenes from the stems of Eurya chinensis.

Two new ent-kaurane diterpenes (1-2), together with five known analogs, were isolated from the stems of Eurya chinensis. The structures of new compounds were established by extensive analysis of mass spectrometric and 1D and 2D NMR spectroscopic data. Compound 3 exhibited noticeable anti-inflammatory activity as denoted by inhibiting LPS-induced nitric oxide (NO) production in RAW264.7 cells with an IC50 value of 7.82 µM. Compound 4 showed potent cytotoxic activity against human cancer cell lines NCI-H46, HepG2 and SW480 with IC50 values ranging from 7.45 to 8.54 µM.

Three new kavalactone dimers from Piper methysticum (kava).

Three new dimeric kavalactones, designated as diyangonins A-C (1-3), along with two known analogs were isolated from the roots of Piper methysticum. Their structures were elucidated by means of extensive analysis of their 1D, 2D NMR, and mass spectroscopic data. All these dimers possess a skeleton featuring a cyclobutane ring connecting two kavalactone units in head-to-tail or head-to-head mode. Compounds 1-5 were evaluated for their cytotoxic activities against human tumor cell lines.

Yangonindimers A-C, three new kavalactone dimers from Piper methysticum (kava).

Three new kavalactone dimers, designated as yangonindimers A-C (1-3), along with one known analogue were isolated from the roots of Piper methysticum. Their structures were elucidated via extensive analysis of their 1D, 2D NMR and mass spectroscopic data. All these dimers possess a skeleton featuring a cyclobutane ring connecting two kavalactone units. Compounds 1-4 were evaluated for their cytotoxic activities against human tumour cell lines NCI-H46, SW480 and HepG2, but none showed significant activity.

MiR-107 down-regulates SIAH1 expression in human breast cancer cells and silencing of miR-107 inhibits tumor growth in a nude mouse model of triple-negative breast cancer.

We have reported that SIAH1 is down-regulated and associated with apoptosis and invasion in human breast cancer. However, the molecular mechanisms leading to SIAH1 down-regulation remain to be elucidated. Here, we demonstrated that miR-107 directly down-regulates SIAH1 expression in human breast cancer cells. Over- expression of miR-107 reduced SIAH1 expression, promoted human breast cancer cell proliferation, colony formation, migration and invasion, and inhibited apoptosis. On the contrary, silencing of miR-107 increased SIAH1 expression and inhibited the tumor growth of MDA-MB-231 cells, a kind of triple-negative breast cancer (TNBC) cells, in vitro and in vivo. Our results reveal that miR-107 is an upstream regulator for SIAH1 down-regulation in human breast cancer cells and miR-107 provides a potential effective target for the treatment of TNBC.

Integrin ß3 and its ligand regulate the expression of uPA through p38 MAPK in breast cancer.

Interplay between integrins and extracellular matrix is suggested to play an important role in malignant progression and tumor differentiation. The aim of the study was to determine the combined expression of integrin ß3 and tenascin-c (TN-c) in breast cancer and examine whether integrin ß3 and TN-c can activate urokinase-type plasminogen activator (uPA) through p38 mitogen-activated protein kinase (p38 MAPK). We detected the expression of integrin ß3, TN-c, p-p38, and uPA in 80 cases of breast invasive ductal carcinoma by immunohistochemistry. In addition, we blocked integrin ß3 and TN-c in the MDA-MB-231 breast cancer cells and detected the expression of p-p38 and uPA by Western blot. Integrin ß3, TN-c, p-p38, and uPA showed high levels of expression in breast invasive ductal carcinoma. The expression of integrin ß3, TN-c, and uPA was correlated with lymph node metastasis and TNM stage in breast cancer. Furthermore, correlations were noted between any two of the three proteins. The expression of p-p38 and uPA decreased in MDA-MB-231 cells after the addition of integrin ß3 antibody and TN-c antibody. The expression of uPA decreased after addition of SB203580. Our results demonstrate that inhibition of the expression of integrin ß3 and TN-c could decrease the expression of uPA through p38 MAPK in breast cancer, suggesting that the interaction between integrin ß3 and TN-c serves an important role in breast cancer.

Expression and regulation mechanisms of Sonic Hedgehog in breast cancer.

Sonic Hedgehog (Shh) plays an essential role in vertebrate organogenesis as well as the development of some cancers, including breast cancer. The aim of the present study was to characterize more precisely its role in breast carcinogenesis and elucidate its regulation mechanisms. The expression of Shh was investigated in 97 breast carcinomas and 22 paired non-tumorous tissues (distant from the primary tumor) by immunohistochemistry and in four breast cell lines by Western blotting. We also analyzed the methylation status of the Shh gene with methylation-specific PCR and assessed whether nuclear factor-kappa B (NF-kappaB) and Gli1 were expressed in breast tissues by immunohistochemistry. Our results showed that Shh protein expression in breast carcinomas was significant higher than that in normal breast tissues (P < 0.01). The up-regulation of Shh in breast carcinomas was correlated significantly with early clinical stage (P < 0.05). In addition, we found a substantial increase in Shh expression at both the mRNA and protein levels in several human breast carcinoma cell lines. The expression level of nuclear Gli1 was positively associated with the expression level of Shh in breast tissues (P < 0.001). Promoter region hypomethylation (43/61, 70.5%) was frequently observed in breast carcinomas and significantly associated with Shh up-regulation (P < 0.05). The DNA methyltransferase inhibitor 5-azacytidine (5-Aza) reduced the methylation of Shh promoter and increased the expression of Shh protein in MDA-MB-435 and MCF-10A cells. Furthermore, most of the breast carcinoma cases with Shh up-regulation had increased expression of NF-kappaB (35/49, 71.4%; P < 0.001). Taken together, these observations suggest that Shh overexpression is a critical event in breast carcinogenesis, and Shh promoter hypomethylation and NF-kappaB up-regulation are responsible for the up-regulation of Shh.

The expression of SIAH1 is downregulated and associated with Bim and apoptosis in human breast cancer tissues and cells.

Seven in absentia homolog1 (SIAH1) was reported as a tumor suppressor and played an important role in regulating cell apoptosis. However, its effects on the breast carcinogenesis remain unclear. In this study, our aims were to examine the relationship between SIAH1 and Bcl-2-interacting mediator of cell death (Bim) and to explore the effects of SIAH1 on the breast carcinogenesis. Immunohistochemical analysis in 231 cases of breast tissues showed that the expression of SIAH1 and Bim were significantly decreased in the breast carcinogenesis. Moreover, SIAH1 expression was significantly correlated with Bim. Both SIAH1 and Bim expression were significantly higher in well to moderately differentiated and in early-stage breast cancer. Reverse transcription (RT)-polymerase chain reaction (PCR) and Western blot analysis in paired breast cancer tissues and breast cell lines found that the expression of SIAH1 was lower in the breast cancer tissues and cell lines. SIAH1 inducing apoptosis of the breast cancer cells was dependent on Bim. However, SIAH1 inhibiting invasion of the breast cancer cells was independent of Bim. The increase of the function of SIAH1 to upregulate the expression of Bim may play an important role in the progression of breast cancer. Restoration of the function of SIAH1 may be a new therapeutic target of human breast cancer.

SIAH1 induced apoptosis by activation of the JNK pathway and inhibited invasion by inactivation of the ERK pathway in breast cancer cells.

Seven in absentia homolog 1 (SIAH1), a homologue of Drosophila seven in absentia (Sina), has emerged as a tumor suppressor and plays an important role in regulating cell apoptosis. To investigate the role and possible mechanism of SIAH1 in breast cancer cells, we up-regulated the expression of SIAH1 using pcDNA3-myc-SIAH1 and knocked down SIAH1 using SIAH1 siRNA. We found that the overexpression of SIAH1 induced cell apoptosis by up-regulating the level of Bim through the activation of the JNK signaling pathway, and the suppression of SIAH1 expression increased cell invasion via the activation of the ERK signaling pathway in breast cancer cells. All these results indicate that the JNK and ERK signaling pathways may play an important role in the SIAH1-dependent biological behavior of breast cancer, and it may be a good molecular therapeutic target to increase the expression level of SIAH1 through promoting cell apoptosis and inhibiting cell invasion in human breast cancer.

[Correlation of p38 mitogen-activated protein kinase signal transduction pathway to uPA expression in breast cancer].

BACKGROUND & OBJECTIVE: p38 mitogen-activated protein kinase (p38MAPK) signal transduction pathway regulates the expression of various metastasis-related genes. Urokinase-type plasminogen activator (uPA) plays an important role in cancer invasion and metastasis. This study was to explore the correlation of p38MAPK signal transduction pathway to uPA expression in breast cancer. METHODS: SP immunohistochemistry was used to test the expression of p-p38, p-Akt, p-Erk, and uPA in 60 specimens of breast cancer. Western blot was used to detect the protein levels of p-p38 and uPA in breast cancer MDA-MB-231 and MCF-7 cells and the protein level of uPA in MDA-MB-231 cells after blocking p38MAPK signal transduction pathway by SB203580, a specific inhibitor of p38MAPK. The correlations of p-p38, p-Akt, p-Erk, and uPA expression to the clinicopathologic characteristics of breast cancer, and the correlations of p-p38, p-Akt, and p-Erk expression to uPA expression were analyzed. The mechanism of p38MAPK signal transduction pathway regulating the protein expression of uPA in breast cancer cells was investigated. The correlation of p-p38 and uPA expression to prognosis of breast cancer was analyzed. RESULTS: The positive rates of p-p38, p-Akt, p-Erk, and uPA proteins in breast cancer tissues were 56.7%, 95.0%, 93.3%, and 60.0%, respectively. The expression of p-p38 was positively correlated to the expression of uPA (r=0.316, P<0.05), while the expression of p-Akt and p-Erk was not related to uPA expression. The expression of p-p38 and uPA was correlated to lymph node metastasis and TNM stage (P<0.05), but not to patients' age and tumor size (P>0.05). The expression of p-Akt and p-Erk were correlated to lymph node metastasis (P<0.05), but not to TNM stage, patients' age, and tumor size (P>0.05). The protein levels of p-p38 and uPA in MDA-MB-231 cells were higher than that in MCF-7 cells. SB203580 concentration-dependently inhibited p38MAPK pathway and induced uPA protein expression. The expression of p-p38 and uPA was negatively correlated to prognosis of breast cancer (log-rank=4.98, 5.40, P=0.026, 0.020). CONCLUSIONS: p38MAPK signal transduction pathway might improve breast cancer progression by up-regulating uPA expression, and might be an important route in invasion and metastasis of breast cancer. p-p38 and uPA might help to evaluate prognosis of breast cancer.

[Relationship between expressions of telomerase genes and apoptosis related genes in mammary ductal atypical hyperplasia].

BACKGROUND & OBJECTIVE: Recent studies showed that the activating of telomerase and excessive expression of apoptosis suppressor genes were related to the development of many tumors. This study was designed to investigate the expression of telomerase genes (hTR, hTRT) and apoptosis related genes (p53, bcl-2) in mammary atypical ductal hyperplasia for exploring the change of telomerase activity and apoptosis related genes in the process of mammary ductal dysplasia to malignant transformation. METHOD: Expressions of telomerase genes (hTR, hTRT) and apoptosis related genes (p53, bcl-2) were detected by in situ hybridization and expression of the mutant p53 protein was detected by immunohistochemistry were detected in 44 patients with mammary atypical ductal hyperplasia, those expression were compared with those of the 6 benign hyperplasia and 26 breast carcinoma. RESULT: High expressions of telomerase genes (hTR, hTRT mRNA) in severe atypical ductal hyperplasia (60.9%, 52.1%) is of significantly difference from that in mild-medium atypical ductal hyperplasia (22.2%, 11.1%; 33.3%, 25.0%) and breast cancer (88.5%, 80.8%). The upgrading of atypia link with decreased expression of wild p53 mRNA (mild: 55.6%; medium: 41.7%; severe: 26.1%) and increased expression of the mutant p53 protein (mild: 11.1%; medium: 25.0%; severe: 34.8%). As for bcl-2 mRNA, it shows moderate expression, especially in severe atypical ductal hyperplasia. CONCLUSION: Our study revealed significant correlation between expressions of telomerase genes (hTR, hTRT) and the state of malignant transformation in mammary atypical ductal hyperplasia. Decreased expression of wild p53 gene, increased expression of mutant p53 protein, and overexpression of bcl-2 gene were associated with telomerase.