GSK-3ß controls autophagy by modulating LKB1-AMPK pathway in prostate cancer cells.

BACKGROUND: Glycogen synthase kinase 3ß (GSK3B, GSK-3ß) is a multi-functional protein kinase involved in various cellular processes and its activity elevates after serum deprivation. We have shown that inhibition of GSK-3ß activity triggered a profound autophagic response and subsequent necrotic cell death after serum deprivation in prostate cancer cells. In this study, we dissected the mechanisms involved in GSK-3ß inhibition-triggered autophagy. METHODS: Prostate cancer PC-3 and DU145 cells were used in the study. Multiple GSK-3ß specific inhibitors were used including small chemicals TDZD8, Tideglusib, TWS119, and peptide L803-mts. Western blot assay coupled with phospho-specific antibodies were used in detecting signal pathway activation. ATP levels were assessed with ATPLite kit and HPLC methods. Autophagy response was determined by evaluating Microtubule-associated proteins 1A/1B light chain 3B (LC3B) processing and p62 protein stability in Western blot assays. Immunofluorescent microscopy was used to detect LKB1 translocation. RESULTS: Inhibition of GSK-3ß activity resulted in a significant decline of cellular ATP production, leading to a significant increase of AMP/ATP ratio, a strong trigger of AMP-activated protein kinase (AMPK) activation in prostate cancer PC-3 cells. In parallel with increased LC-3B biosynthesis and p62 protein reduction, the classical sign of autophagy induction, AMPK was activated after inhibition of GSK-3ß activity. Further analysis revealed that Liver kinase B1 (LKB1) but not Calcium/calmodulin-dependent protein kinase kinase ß (CaMKKß) is involved in AMPK activation and autophagy induction triggered by GSK-3ß inhibition. Meanwhile, GSK-3ß inhibition promoted LKB1 translocation from nuclear to cytoplasmic compartment and enhanced LKB1 interaction with its regulatory partners Mouse protein-25 (MO25) and STE20-related adaptor (STRAD). CONCLUSIONS: In conclusion, our data suggest that GSK-3ß plays an important role in controlling autophagy induction by modulating the activation of LKB1-AMPK pathway after serum deprivation.

Two novel phenanthraquinones with anti-cancer activity isolated from Bletilla striata.

7-Hydroxy-2-methoxy-phenanthrene-3,4-dione and 3',7',7-trihydroxy-2,2',4'-trimethoxy-[1,8'-biphenanthrene]-3,4-dione, two novel compounds and four known compounds were isolated from Bletilla striata. The structures of the compounds were established on the basis of extensive spectroscopic analysis. The two compounds exhibited antiproliferative effects using the MTT test; these effects may be due to cell cycle arrest and inducing ROS generation.

Biological Evaluation of Ferrocenyl Olefins: Cancer Cell Growth Inhibition, ROS Production, and Apoptosis Activity.

The antiproliferative effects of various ferrocenyl olefins were evaluated against the cell lines MCF-7 (human breast cancer cells), DLD-1 (human colon adenocarcinoma cells), HUVEC (human umbilical vein endothelial cells), and A549 (human lung carcinoma cells), using the MTT test. IC50 values were determined. Compounds 8, 9, 11, and 12 with high antiproliferative activity were tested for their reactive oxygen species (ROS) production, and cell cycle analysis was performed on A549 cells. The results show that these compounds might perform their antiproliferative activity through inducing ROS generation, apoptosis induction, and cell cycle arrest.

Neuroprotection by saponins.

Saponins, an important group of bioactive plant natural products, are glycosides of triterpenoid or steroidal aglycones. Their diverse biological activities are ascribed to their different structures. Saponins have long been recognized as key ingredients in traditional Chinese medicine. Accumulated evidence suggests that saponins have significant neuroprotective effects on attenuation of central nervous system disorders, such as stroke, Alzheimer's disease, Parkinson's disease, and Huntington's disease. However, our understanding of the mechanisms underlying the observed effects remains incomplete. Based on recently reported data from basic and clinical studies, this review highlights the proposed mechanisms of their neuroprotective function including antioxidant, modulation of neurotransmitters, anti-apoptosis, anti-inflammation, attenuating Ca(2+) influx, modulating neurotrophic factors, inhibiting tau phosphorylation, and regeneration of neural networks.

[Clinicopathologic features of inflammatory fibroid polyp in gastrointestinal tract].

OBJECTIVE: To study the clinicopathologic characteristics, immunophenotypes, pathologic diagnosis and differential diagnosis of gastrointestinal inflammatory fibroid polyp (IFP). METHODS: The clinical data, histologic features and immunohistochemical results of 6 cases of gastrointestinal inflammatory fibroid polyp were analyzed, with review of literature. RESULTS: There were altogether 5 women and a man. The age of patients ranged from 44 to 87 years (median age = 61 years and mean age = 62 years). The sites of involvement included stomach (number = 2), jejunum (number = 2), distal ileum (number = 1) and rectum (number = 1). The patients often presented with abdominal pain or discomfort and tarry stool. All the cases studied showed similar morphology, were submucosa-based and composed of bland spindle cells associated with loose collagenous stroma and perivascular edema. Prominent concentric fibroblastic proliferation ("onion-skin" appearance) and eosinophilic infiltrate were characteristics. Three of the patients underwent surgical resection. The remaining patients had the lesions removed with endoscopy. Follow-up data were available in 5 patients and none of them had disease recurrence or metastasis. CONCLUSIONS: IFP is a rare benign tumor of gastrointestinal tract. Correct diagnosis primarily relies on pathologic examination. It needs to be distinguished from gastrointestinal stromal tumors or inflammatory myofibroblastic tumor.

Expression status of fatty acid synthase (FAS) but not HER2 is correlated with the differentiation grade and prognosis of esophageal carcinoma.

BACKGROUND/AIMS: Fatty acid synthase (FAS) and human epidermal growth factor receptor 2 (HER2) are overexpressed in a series of cancers. However, few studies have investigated the potential role of FAS and HER2 in esophageal carcinoma. The aim of this study was to evaluate the expression of FAS and HER2 and the possible link between FAS/HER2 expression and the pathological prognostic variables. METHODOLOGY: The frequency of FAS and HER2 expression was determined immunohistochemically. The overall survival rate was analysed by the Kaplan-Meier method and the log-rank test using SPSS 17.0 software. RESULTS: The majority of the cases were esophageal squamous cell carcinomas (n=142). FAS and HER2 overexpression in the studied cases are 73.2% and 14.1%, respectively. There was a significant difference in FAS expression regarding tumor differentiation and FAS overexpression showed its prognostic value for patients with different tumor differentiation. Meanwhile, HER2 overexpression did not significantly relate to the clinicopathological characteristics of the tumors, with the only exception of the surgical margins. CONCLUSIONS: FAS and HER2 overexpression are common in esophageal carcinomas. FAS overexpression showed its prognostic value for esophageal carcinoma patients with different tumor differentiation, which lead us to consider FAS-inhibitors as potential candidates for target-based adjuvant therapies.

[A comparative study of laparoscopic-assisted and open distal gastrectomy for advanced gastric cancer].

OBJECTIVE: To evaluate the safety, feasibility and the long-term outcomes of laparoscopy-assisted gastrectomy (LAG) for advanced gastric cancer (AGC). METHODS: The clinical and follow-up data of 46 cases after LAG from June 2008 to December 2009 were analyzed, and compared with 85 cases after conventional open gastrectomy (OG) for advanced gastric cancer at the same period at our hospital. The surgical safety, postoperative recovery, complications, radical degree, survival rate were compared. RESULT: As compared with OG group, operation time was longer in LATG group ((274 ± 78) min vs. ( 217 ± 41) min, t = 4.635, P = 0.000). Estimated blood loss in the LAG group ((254 ± 112) ml) was significantly less than in the OG group (t = 3.942, P = 0.000). Time to ambulation ((63 ± 16) hours), first flatus ((77 ± 20) hours), resumed liquid diet ((88 ± 15) hours), duration of analgesic medication ((53 ± 20) hours) and postoperative hospital stay ((11.1 ± 4.6) days) were significantly shorter in the LAG group (t = 5.549, 6.508, 9.436, 9.464 and 2.980 respectively, all P < 0.01). The distance of the proximal and distal resection margin were (5.7 ± 1.4) cm and (3.9 ± 1.5) cm in LAG group, (5.8 ± 1.1) cm and (4.7 ± 1.5) cm in OG group respectively, but the difference was not significant. The number of lymph node dissections was also similar, (30.5 ± 10.4) in LAG group and (32.6 ± 12.3) in OG group (t = 0.960, P = 0.339). The incidence of postoperative complications and mortality rate in LAG group (8.7% and 0 respectively) were also lower than in the OG group, with no statistically significant difference (P > 0.05). The mean follow-up was 31.0 months (range 6-48 months), and the cumulative survival of the 2 groups was similar (χ(2) = 1.594, P = 0.207). CONCLUSIONS: Laparoscopy-assisted gastrectomy for advanced gastric cancer is not significantly different with open surgery in surgical safety, radical degree, and survival rate. It is less traumatic and of fewer complications.

Design of cationic surfactant reinforced carrageenan waterproof composite films and applied as water induced electricity generator.

Carrageenan (CR) is a renewable polysaccharide material for packaging application due to its good film-forming property, but its use can be limited by the water solubility. In this research, CR hydrogels were modified by quaternary ammonium surfactants with different length of hydrocarbon tails (n, 8≦n≦16) by adsorption method and waterproof films were obtained after drying. The composition and charge interaction of composite films was confirmed by FTIR. Both thermogravimetric analysis and energy dispersive spectrometer indicated that the surfactant ions replaced K+ to form complexes with CR. The X-ray diffraction revealed the decreased amorphous nature of composite films compared to neat CR film. Water-related physical properties, such as water content, weight percentage change after contact with water, water vapor transmission, and water contact angle were intimately related to n. When 8≦n≦14, the waterproof properties were enhanced with the increase of n. Meanwhile, the waterproof property of composite film was ascertained by the no leakage result in the boiling water packaging experiment. When n = 16, sandwich structure was found in the sectional micromorphology images, and water bag structure formed after immersed into water. By comparing the mechanical properties of the composite films in different condition, we found that quaternary ammonium surfactants improved significantly the tensile strength in water and increased elongation at break in dry state. The composite films can be used as water induced voltage generator for their polyelectrolyte nature. Benefiting from the high stability of the composite films in water, their water-induced voltage generation process had good recyclability. Due to the antimicrobial activity of the quaternary ammonium salts and the waterproof property, composite films were more stable and degraded more slowly than neat CR film in nature environment.

[Research progresses of plant polysaccharides on cell adhesion].

Cell adhesion mediated by cell adhesion molecules (CAMs) constitutes essential life phenomenon. In inflammation, immunity, infection, thrombosis, tumor metastasis and wound healing, cell adhesion comes into being the basic physiological and pathological process. Intervening with cell adhesion has been the important therapeutic and prophylactic strategies for diseases. Accumulated evidence has indicated that plant polysaccharides especially those exacted from Chinese traditional and herbal drugs displayed various pharmacological effects such as anti-inflammation, anti-cancer, anti-infection, immunomodulation, cardiovascular protective effects and so on. In this paper, the research progress of plant polysaccharides on cell adhesion is reviewed.

[LiCl suppresses tumor growth and inhibits DNA replication in prostate cancer].

OBJECTIVE: To assess the effects of LiCl on prostate cancer growth and to explore the underlying mechanisms. METHODS: Effects of LiCl on cell growth in vitro and in vivo were determined by cell counting and xenografts of prostate cancer cells. Alterations in cell proliferation and the expression of DNA replication-related protein were determined by MTT assay, BrdU incorporation and Western blot. RESULTS: Compared to PBS control group, the number of prostate cancer cells (PC-3) were lower treated with 10 mmol/L LiCl, the number was 1.9×10(5), 4.8×10(5) and the difference was significant (P < 0.05). The inhibition rate of cellular proliferation were 50%, 95% and 98%, respectively, in LiCl group, NaCl and KCl control group, the difference was significant (P < 0.05). The A-Value of BrdU incorporation was 1.5, 1.3 treated with 10 mmol/L, 30 mmol/L LiCl, while the A-value of BrdU incorporation was 4 in PBS control group, the difference was significant (P < 0.05). On the protein level, LiCl downregulates expression of cdc 6, cyclins A and cyclins E, and cdc 25C, and upregulates expression of the CDK inhibitor p21(CIP1). The mean volume and weight of xenograft tumor were 50 mm(3) and 296 mg after LiCl intraperitoneal injection, But PBS control group were 180 mm(3) and 957 mg, the difference was significant (P < 0.05). CONCLUSION: LiCl disrupts DNA replication and suppresses tumor growth of prostate cancer cells in vitro and in vivo.

[Gastrointestinal adenocarcinomas with a micropapillary pattern: a clinicopathologic and immunohistochemical study].

OBJECTIVE: To study the clinicopathologic features and immunophenotype of gastrointestinal adenocarcinomas with a micropapillary pattern differentiation (GAMPD). METHODS: Seventy-three cases of GAMPD arising in gastrointestinal tract were retrospectively reviewed. Immunohistochemical study for epithelial membrane antigen (EMA), insulin-like growth factor II mRNA-binding protein-3 (IMP3) and E-cadherin was performed. RESULTS: Amongst the 73 cases studied, the micropapillary pattern accounted for 5% to 70% of the tumor component. It was often seen in a background of moderately differentiated adenocarcinoma. As compared with conventional adenocarcinoma, nodal metastasis was more frequently observed and the TNM tumor stage was statistically higher in GAMPD. The occurrence of micropapillary component in metastatic lymph nodes positively correlated with the proportion of micropapillary pattern in primary lesions. EMA staining on the stroma-facing surface of tumor micropapillae was demonstrated in 52.1% (38/73) of the cases. As compared with EMA-negative GAMPD, EMA-positive GAMPD was more in the stomach (P = 0.018), and with more metastatic lymph nodes (6.6 ± 5.8 vs 3.8 ± 4.7, P = 0.029). The rate of IMP3 expression in EMA-positive GAMPD was 86.8%(33/38), which was higher than that in conventional adenocarcinoma. In contrast, the rate of E-cadherin expression in GAMPD was lower than that in conventional adenocarcinoma. CONCLUSIONS: GAMPD is a distinctive variant of gastrointestinal adenocarcinomas and different from conventional adenocarcinoma in tumor morphology, immunophenotype and biologic behavior. It carries an aggressive clinical course and poor prognostic outcome. Immunohistochemical study for EMA, IMP3 and E-cadherin would be helpful in the diagnosis and prognostic evaluation of GAMPD.

Adeno-associated virus-delivered short hairpin-structured RNA for androgen receptor gene silencing induces tumor eradication of prostate cancer xenografts in nude mice: a preclinical study.

The androgen receptor (AR) is the most critical factor in prostate cancer progression. We previously demonstrated that silencing the AR using 2 unique small interfering RNAs (no. 8 and no. 31 AR siRNA) induces apoptotic cell death in AR-positive prostate cancer cells. To develop this AR siRNA technique into a therapy for prostate cancers, we generated an adeno-associated virus (AAV) vector to stably express a short hairpin-structured RNA (shRNA) against the AR gene in vivo. In addition to the no. 8 AR shRNA (ARHP8), we also screened a group of AR shRNAs with different sequences and identified a less effective AR shRNA (ARHP4) that was used as an shRNA control. An empty AAV vector (AAV-GFP) was used as a negative control. Intratumoral injection of AAV-ARHP8 viruses significantly suppressed tumor growth of xenografts derived from either androgen-responsive or castration-resistant prostate cancer cells. Most interestingly, systemic delivery of the AAV-ARHP8 but not AAV-ARHP4 or AAV-GFP viruses via tail vein injection eliminated xenografts within 10 days. Further analysis revealed that AAV-ARHP8 viruses dramatically reduced the expression of AR-regulated cellular survival genes and caused a dramatic apoptotic response. Taken together, our data strongly suggest that AAV-ARHP8 viruses induced a strong AR gene silencing in vivo and that systemic delivery of ARHP8 siRNA via an AAV vector or any other means might be considered as novel gene therapy for prostate cancers.

[Expression of BRG1 and BRM proteins in prostatic cancer].

OBJECTIVE: To study the effect of BRG1 and BRM, the catalytic subunits expressed by SWI/SNF, in benign and malignant prostatic tissues and to correlate the BRG1/BRM expression with the development and progression of prostatic cancer. METHODS: The expression levels of the BRG1 and BRM proteins in benign and malignant prostatic tissues were studied using semi-quantitative immunohisto-chemistry. The results correlated with various clinical and pathologic parameters. RESULTS: The average immuno-reactive score for BRG1 expression in prostatic cancer tissues was significantly higher than that in benign prostatic tissues (57+/-9.8 and 19+/-4.1, respectively, P = 0.000 17). The difference was more obvious in the high-grade cancer. On the other hand, BRM expression exhibited a heterogeneous pattern. The average immuno-reactive score for BRM expression was lower in cancer tissues than in benign tissues (112+/-17 and 151+/-19, respectively, P = 0.0047). BRG1 and BRM demonstrated a reciprocal expression pattern in benign and malignant tissues. The average immuno-reactive score for BRG1 expression was higher in the cancer cases with a larger tumor volume than in the cases with a smaller tumor volume (P = 0.0112). CONCLUSIONS: The expression of BRG1 and BRM correlates with the development of prostatic cancer. Increased BRG1 expression may have certain implications in tumor progression.