STK10 mutations block erythropoiesis in acquired pure red cell aplasia via impairing ribosome biogenesis.

Acquired pure red cell aplasia (PRCA) is anemia associated with the absence of erythroblasts and is characterized by persistent and easy recurrence. However, the underlying mechanisms of acquired PRCA remain obscure, and the role of gene mutations in the pathogenesis of acquired PRCA is not fully characterized. In the present study, we detected thirty newly diagnosed patients with acquired PRCA using whole exome sequencing, and a potential role for STK10 in acquired PRCA was uncovered. The mRNA levels of STK10 in three patients with STK10 mutations were decreased. These three patients had a poor response to immunosuppressive therapy and two died in the follow-up period. Here we report that knockdown of STK10 inhibits erythroid differentiation and promotes apoptosis of K562 cells. We show that knockdown of STK10 resulted in inhibition of ribosome biogenesis and reduced ribosome levels in K562 cells. We also show that the p53 signaling pathway is activated by knockdown of STK10. Our results imply that ribosome biogenesis downregulation together with pathological p53 activation prevents normal erythropoiesis. Our study uncovers a new pathophysiological mechanism leading to acquired PRCA driven by STK10 mutations.

Higher systolic blood pressure is specifically associated with better islet beta-cell function in T2DM patients with high glycemic level.

BACKGROUND: Patients with type 2 diabetes mellitus (T2DM) usually have higher blood viscosity attributed to high blood glucose that can decrease blood supply to the pancreas. A mild increase in blood pressure (BP) has been reported as a potential compensatory response that can maintain blood perfusion in the islet. However, how BP influences beta-cell function in T2DM subjects remains inconsistent. This study aimed to examine the relationship between BP and beta-cell function in patients with T2DM under different HbA1c levels. METHODS: This is a cross-sectional study of 615 T2DM patients, whose clinical data were extracted from hospital medical records. Beta-cell function was assessed by insulin secretion-sensitivity index-2 (ISSI2). Multivariable linear regression analysis and restricted cubic splines (RCS) analysis were performed to identify the association between systolic BP (SBP) and ISSI2. Mediation analysis was performed to determine whether higher SBP could reduce blood glucose by enhancing beta-cell function. RESULTS: After adjustment of potential confounders, in participants with HbA1c ≥ 10%, the SBP between 140 to150 mmHg had the highest log ISSI2 (b = 0.227, 95% CI 0.053-0.402), an association specific to participants with < 1 year duration of diabetes. RCS analyses demonstrated an inverted U-shaped association between SBP and ISSI2 with the SBP at 144 mmHg corresponding to the best beta-cell function. This higher SBP was "paradoxically" associated with lower 2 h postprandial blood glucose (PBG) when SBP < 150 mmHg that was almost exclusively mediated by ISSI2 (mediating effect = - 0.043, 95%CI - 0.067 to - 0.018; mediating effect percentage = 94.7%, P < 0.01). SBP was however not associated with improvement in ISSI2 or 2 h PBG in participants with HbA1c < 10%. CONCLUSIONS: In early stage of diabetes, a slightly elevated SBP (140-150 mmHg) was transiently associated with better beta-cell function in T2DM patients with HbA1c ≥ 10% but not in those with HbA1c < 10%.

Efficacy and Safety of Concentrated Growth Factor Fibrin on the Extraction of Mandibular Third Molars: A Prospective, Randomized, Double-Blind Controlled Clinical Study.

PURPOSE: To investigate the efficacy and safety of concentrated growth factor fibrin (CGF) for the extraction of mandibular third molars. PATIENTS AND METHODS: This was a randomized, double-blind, and controlled clinical study. Patients who underwent mandibular impacted tooth extraction were randomly divided into 2 groups. In the CGF group, the tooth extraction fossa was utilized to place CGF gel. In the control group, the fossa was filled with serum. The visual analogue scale (VAS), reductions in swelling and trismus, incidence of postoperative dry socket, distal periodontal depth and bone regeneration of the second molar, and bone density (BMD) of the extraction fossa at 24 weeks were evaluated. RESULTS: One hundred eighteen patients were enrolled in this study. There was no significant difference in baseline clinical characteristics between the 2 groups. The pain score of the CGF group was significantly lower than that of the control group at 2, 24, and 48 hours after operation. There was no significant difference in the reduction in swelling or trismus between the 2 groups. There were no cases of dry socket in the CGF group and 3 cases of dry socket in the control group. The periodontal probing depth and bone regeneration of the second molar when the socket was implanted with CGF were better than those that healed naturally (P < .05). The bone mineral density of each group was significantly increased at 24 weeks but was significantly different between groups (P < .05). CONCLUSION: CGF can effectively reduce reactive tooth extraction pain and help avoid dry sockets. It can promote periodontal tissue and bone healing in distal and extracted sockets.

A peptide coating preventing the attachment of Porphyromonas gingivalis on the surfaces of dental implants.

OBJECTIVES: The aim of this study was to investigate whether a peptide-based coating can prevent the adhesion of Porphyromonas gingivalis, a key human pathogen associated with periodontitis and peri-implantitis. BACKGROUND: Nonsurgical and surgical interventions have been used for the treatment of peri-implantitis; however, the effectiveness of these approaches is usually unsatisfactory. The main reason is that dental plaque on the surface of the implant is difficult to remove due to its rough surface and thread design. Recently, a peptide-based coating for implant surfaces that can reject the adhesion of Escherichia coli and improve the attachment of host cells was developed. METHODS: A salivary pellicle was created on the surfaces of peptide-coated bare discs and verified with anti-human immunoglobulin G, A and M, and anti-fibrinogen. Early colonizers, Veillonella parvula and Streptococcus sobrinus, and the later colonizer, Porphyromonas gingivalis, were labelled with green and red fluorescent dyes, respectively, and seeded on the discs. Bacterial attachment was semi-quantified by fluorescence intensity. RESULTS: The salivary pellicle was evenly distributed on the discs, with or without the peptide coating, with an average thickness of 3.84 µm. A multi-species dental biofilm was created on the salivary pellicle. The peptide coating resulted in an approximate 25% reduction in the attachment of Veillonella parvula and Streptococcus sobrinus, and a 50% reduction in Porphyromonas gingivalis, when compared to control, uncoated implant discs. CONCLUSION: The novel peptide-based coating can inhibit the attachment of Porphyromonas gingivalis. It may have the potential to impede the development of peri-implantitis.

Clinical Application of Concentrated Growth Factor Fibrin Combined With Bone Repair Materials in Jaw Defects.

PURPOSE: We investigated the promoting effect of concentrated growth factor (CGF) fibrin on the repair of jaw bone defects. PATIENTS AND METHODS: We designed a clinical trial composed of patients with jaw defects. Forty patients were divided into the test and control groups. CGF fibrin combined with Bio-Oss bone powder (Giestlich Pharma, Wolhusen, Switzerland) was used in the test group. Bio-Oss bone powder alone was used in the control group. The concentration of vascular endothelial growth factor (VEGF) and transforming growth factor (TGF)-ß in the red blood cell (RBC) layer and CGF gel was measured. At different time points before and after surgery, the serum bone alkaline phosphatase (BAP), osteocalcin, and bone mineral density levels were measured. Regular examinations and computed tomography scans were also performed in the follow-up period. RESULTS: The CGF fibrin available for clinical use was obtained by centrifugation. One day after preparation by centrifugation, the VEGF and TGF-ß concentration in the CGF gel was 2.57-fold and 3.4-fold greater than the concentration in the RBC layer, respectively. The BAP and osteocalcin levels increased at 1 and 12 weeks postoperatively in both groups. Furthermore, the BAP and osteocalcin levels in the test group were significantly greater than those in the control group at 1 and 12 weeks postoperatively (P < .05 for all). The bone mineral density in the bone defect area of the test group was also significantly greater than that of the control group at 6 months postoperatively (P < .05). Evaluation of the regular radiographic scans revealed that the effects in the test group were better than those in the control group. CONCLUSIONS: CGF fibrin could promote new bone formation in jaw defects, with benefit to the healing of bone tissue and, thus, is a promising bone repair material.

Isoliquiritigenin Inhibits Ovarian Cancer Metastasis by Reversing Epithelial-to-Mesenchymal Transition.

The epithelial-to-mesenchymal transition (EMT) plays a prominent role in cancer metastasis. Isoliquiritigenin (ISL), one of the flavonoids in licorice, has been shown to exhibit anticancer activities in many cancer types through various mechanisms. However, it is unknown whether ISL impacts the EMT process. Here, we show that ISL is able to suppress mesenchymal features of ovarian cancer SKOV3 and OVCAR5 cells, evidenced by an apparent morphological change from a mesenchymal to an epithelial phenotype and reduced levels of mesenchymal markers accompanied by the gain of E-cadherin expression. The suppression of EMT is also supported by the observed decrease in cell migration and in vitro invasion upon ISL treatment. Moreover, we show that ISL effectively blocks the intraperitoneal xenograft development of the SKOV3 cell line and prolonged the survival of tumor-bearing mice. These data suggest that ISL inhibits intraperitoneal ovary tumor development through the suppression of EMT, indicating that ISL may be an effective therapeutic agent against ovarian cancer.

An immune and epigenetics-related scoring model and drug candidate prediction for hepatic carcinogenesis via dynamic network biomarker analysis and connectivity mapping.

Hepatocellular carcinoma (HCC) is a malignant tumor with high mortality. This study aimed to build a prognostic signature for HCC patients based on immune-related genes (IRGs) and epigenetics-related genes (EPGs). RNA-seq data from Gene Expression Omnibus were used for dynamic network biomarker (DNB) analysis to identify 56 candidate IRG-EPG-DNBs and their first-neighbor genes. These genes were screened using LASSO-Cox regression analysis to finally obtain five candidate genes-RNF2, YBX1, EZH2, CAD, and PSMD1-which constituted the prognostic signature panel. According to this panel, patients in The Cancer Genome Atlas and International Cancer Genome Consortium were divided into high- and low-risk groups. The prognosis, clinicopathological features, and immune cell infiltration significantly differed between the two risk groups. The prognostic ability of the signature panel and expression profiling were further validated using online databases. We used an independent cohort of patients to validate the expression profiles of the five genes using reverse transcription-PCR. CMap and CellMiner predicted four small molecule drug-protein pairs based on the five prognostic genes. Of them, two market drugs approved by the Food and Drug Administration (AT-13387 and KU-55933) have emerged as candidates for HCC study. This new signature panel may serve as a potential prognostic marker, engendering the possibility of novel personalized therapy with classification of HCC patients.

Shenqi Fuzheng injection reverses M2 macrophage-mediated cisplatin resistance through the PI3K pathway in breast cancer.

BACKGROUND: Shenqi Fuzheng injection (SQFZ) combined with chemotherapy can sensitize tumour cells. However, the mechanisms underlying SQFZ's effects remain unknown. In human breast cancer cell lines and M2 macrophages, we showed that SQFZ was a significantly potent agent of sensitization. METHODS: The human breast cancer cell line, MDA-MB-231/DDP, and the human acute leukaemia mononuclear cell line, THP-1, were used. MDA-MB-231/DDP breast cancer xenografts were established to monitor tumour growth. Resistance-associated proteins were examined by western blotting. Levels of cytokines and chemokines were detected by ELISA. Cell viability was measured using the MTT assay. Apoptosis was detected by flow cytometric analysis. RESULTS: SQFZ significantly enhanced the capability of cisplatin to reduce tumour mass. SQFZ and cisplatin decreased the expression of CD206 by 1.89-fold and increased that of CD86 by 1.76-fold as compared to cisplatin alone. The levels of PGE2, IL-6, and CCL1 decreased significantly, and the activation of p-PI3K and the expressions of P-gp and ABCG2 were also inhibited by SQFZ in combination with cisplatin treatment in vivo. The survival following cisplatin administration of 60 µM and 120 µM reduced significantly in the presence of SQFZ in MDA-MB-231/DDP and M2 co-cultured cells. IGF-1, a PI3K activator, combined with SQFZ weakened the effects of SQFZ-induced apoptosis from 28.7% to 10.5%. The effects of IGF-1 on increasing the expressions of P-gp, ABCG2, and Bcl-2, and decreasing that of Bax were reversed by SQFZ. CONCLUSION: Our findings provide evidence that SQFZ is a potential therapeutic drug for cancer therapy.

[Penehyclidine hydrochloride regulates angiopoietin 2/vascular endothelial cadherin (Ang2/VE-cadherin) pathway to alleviate LPS induced lung injury in rats].

Objective To explore the effect and mechanism of penehyclidine hydrochloride (PHCD) on vascular endothelial injury in septic rats. Methods Fifty male SD rats were randomly divided into control group, lipopolysaccharide (LPS) induced sepsis group (model group), low dose PHCD (0.3 mg/kg) group, medium dose PHCD (1.0 mg/kg) group and high dose PHCD (3.0 mg/kg) groups, ten mice for each group. Normal saline was injected into the tail vein of the control group, and 10 mg/kg lipopolysaccharide (LPS) was injected into the tail vein of the rats in other groups to prepare the sepsis rat models. After the models were successfully established, low, medium and high doses (0.3, 1.0, 3.0 mg/kg) of PHCD solution were injected into the tail vein of the rats of corresponding groups. Wet/dry mass ratio (W/D) of lung tissue of rats in each group was measured, and ELISA was used to assay interleukin-1ß (IL-1ß), tumor necrosis factor-α (TNF-α), IL-6 content and rat plasma angiopoietin 2 (Ang2) content in bronchoalveolar lavage fluid (BALF). HE staining was used to observe the pathological changes of lung tissues. Immunohistochemical staining was used to observe the expression of Ang2 in the right lung tissues. Western blot analysis was performed to detect Ang2 and vascular endothelial cadherin (VE-cadherin) protein in lung tissues. Results Compared with the control group, the W/D ratio of the lung tissues of rats in the model group and the contents of IL-1ß, IL-6 and TNF-α in BALF were significantly increased; the lung tissues showed obvious pathological damage, with up-regulation of Ang2 expression and down-regulation of VE-Cadherin expression. Compared with the model group, the W/D ratio of the lung tissues of rats in three PHCD treatment groups and the contents of IL-1ß, IL-6 and TNF-α in BALF were significantly reduced; the pathological damage of lung tissue was significantly reduced, with down-regulation of Ang2 expression and up-regulation of VE-cadherin expression. Conclusion PHCD can reduce LPS-induced lung inflammation in rats with sepsis by regulating the Ang2/VE-Cadherin pathway, thereby improving vascular endothelial injury.

Chemokine C-C motif receptor 5 and C-C motif ligand 5 promote cancer cell migration under hypoxia.

The chemokine CC motif receptor 5 (CCR5) and its ligands have been reported to be associated with cancer progression and metastasis. Although recent researches have demonstrated a fundamental role of hypoxia in cancer, the effect of hypoxia on the expression and function of CCR5 and its ligands in cancer cells is unknown. Here, we investigated the status of CCR5 and its ligands in cancer cells under hypoxic conditions. Quantitative polymerase chain reaction, western blotting and immunofluorescence staining showed that hypoxia induced a strong increase of CCR5 expression. Dual luciferase assay and mRNA stability analysis indicated that hypoxia-induced CCR5 mRNA expression relied on both transcriptional and posttranscriptional mechanisms. We detected the expression of CCR5 ligands and found that chemokine CC motif ligand 5 (CCL5) was induced under hypoxia. Recombinant human CCL5 stimulated cell migration rather than cell proliferation under hypoxia, and neutralization of CCL5 inhibited hypoxia-induced migration of cancer cells. Similarly, overexpression of CCR5 increased cell migration, and knockdown of CCR5 attenuated hypoxia-mediated cell migration. We further showed that hypoxia-inducible factor-1α (HIF-1α) was involved in CCR5 and CCL5 regulation under hypoxia. HIF-1α mRNA levels were highly correlated with CCR5 mRNA and CCL5 mRNA levels in clinical samples. CCR5 and CCL5 were highly expressed in breast cancer lymph nodes metastases. Taken together, our data suggest that CCR5-CCL5 interaction promotes cancer cell migration under hypoxia.

Dynamic network biomarker analysis and system pharmacology methods to explore the therapeutic effects and targets of Xiaoyaosan against liver cirrhosis.

ETHNOPHARMACOLOGICAL RELEVANCE: Xiaoyaosan is a traditional Chinese herbal formula that has long been used to treat liver cirrhosis, liver failure, and hepatocarcinoma (HCC). However, little is known about its mechanism of action and targets in treating chronic liver disease. AIM OF THE STUDY: This study aimed to detect the critical transition of HCC progression and to explore the regulatory mechanism and targets of Xiaoyaosan treating liver cirrhosis (cirrhosis) using integrative medicinal research involving system biology and pharmacology. MATERIALS AND METHODS: We recruited chronic liver disease participants to obtain gene expression data and applied the dynamic network biomarker (DNB) method to identify molecular markers and the critical transition. We combined network pharmacology and DNB analysis to locate the potential DNBs (targets). Then we validated the DNBs in the liver cirrhosis rat models using Xiaoyaosan treatment. The expression of genes encoding the four DNBs, including Cebpa, Csf1, Egfr, and Il7r, were further validated in rat liver tissue using Western blot analysis. RESULTS: We found EGFR, CEBPA, Csf1, Ccnb1, Rrmm2, C3, Il7r, Ccna2, and Peg10 overlap in the DNB list and Xiaoyaosan-Target-Disease (XTD) network constructed using network pharmacology databases. We investigated the diagnostic ability of each member in the DNB cluster and found EGFR, CEBPA, CSF1, and IL7R had high diagnostic abilities with AUC >0.7 and P-value < 0.05. We validated these findings in rats and found that liver function improved significantly and fibrotic changes were relieved in the Xiaoyaosan treatment group. The expression levels of CSF1 and IL7R in the Xiaoyaosan group were significantly lower than those in the cirrhosis model group. In contrast, CEBPA expression in the Xiaoyaosan group was significantly higher than that in the cirrhosis model group. The expression of EGFR in the Xiaoyaosan group was slightly decreased than in the model group but not significantly. CONCLUSION: Using the DNB method and network pharmacology approach, this study revealed that CEBPA, IL7R, EGFR, and CSF1 expression was remarkably altered in chronic liver disease and thus, may play an important role in driving the progression of cirrhosis. Therefore, CEBPA, IL7R, EGFR, and CSF1 may be important targets of Xiaoyaosan in treating cirrhosis and can be considered for developing novel therapeutics.

Hybrid Pyramid Convolutional Network for Multiscale Face Detection.

Face detection remains a challenging problem due to the high variability of scale and occlusion despite the strong representational power of deep convolutional neural networks and their implicit robustness. To handle hard face detection under extreme circumstances especially tiny faces detection, in this paper, we proposed a multiscale Hybrid Pyramid Convolutional Network (HPCNet), which is a one-stage fully convolutional network. Our HPCNet consists of three newly presented modules: firstly, we designed the Hybrid Dilated Convolution (HDC) module to replace the fully connected layers in VGG16, which enlarges receptive field and reduces its loss of local information; secondly, we constructed the Hybrid Feature Pyramid (HFP) to combine semantic information from higher layers together with details from lower layers; and thirdly, to deal with the problem of occlusion and blurring effectively, we introduced Context Information Extractor (CIE) in HPCNet. In addition, we presented an improved Online Hard Example Mining (OHEM) strategy, which can enhance the average precision of face detection by balancing the number of positive and negative samples. Our method has achieved an accuracy of 0.933, 0.924, and 0.848 on the Easy, Medium, and Hard subset of WIDER FACE, respectively, which surpasses most of the advanced algorithms.

Lingual Linear Lesions: A Clinical Sign Strongly Suggestive of Severe Vitamin B12 Deficiency.

BACKGROUND: Lingual linear lesions (LLLs) are the oral linear lesions located on the dorsum, lateral borders, and/or ventral surface of tongue. It has been suggested that LLLs might be an early clinical sign of vitamin B12 deficiency. Here, a retrospective study was conducted to further investigate and validate the association between LLL and vitamin B12 deficiency. METHODS: Based on the clinical examination, patients with LLLs were enrolled and analyzed retrospectively. Data regarding clinical and laboratory features were obtained. Follow-up was done at least 6 months following appropriate supplementation therapy. RESULTS: A total of 57 patients, consisting of 20 males and 37 females with a mean age of 59.12 years (range, 18-80), were enrolled in this study. The hematological examination revealed that 56 (98.25%) of the 57 patients had severe serum vitamin B12 deficiency, and the other 1 patient had a borderline low level of vitamin B12 . All the enrolled patients responded well to cobalamin replacement therapy. CONCLUSION: LLLs might be a clinical sign strongly suggestive of severe vitamin B12 deficiency.

Anticancer activity of Voacangine against human oral cancer cells is due to G2/M cell cycle arrest, ROS-mediated cell death and inhibition of PI3K/AKT signalling pathway.

PURPOSE: Oral cancer is the 6th most prevalent type of cancer and is responsible for high human morbidity and mortality. The present study was designed to investigate the anticancer effects of Voacangine against human oral cancer and to decipher the underlying molecular mechanisms responsible for its anticancer properties. METHODS: CCC-1 oral cancer cell line and normal hTRET-OME cell line were used in this study. Cell viability was determined by MTT assay. Acridine orange (AO)/ ethidium bromide (EB) and annexin V/propidium iodide (PI) assay were used for assessment of apoptosis. Cell cycle analysis and reactive oxygen species (ROS) determination was done by flow cytometry. The protein expression was determined by western blot analysis. RESULTS: The results showed that Voacangine caused a remarkable decline in proliferation of SCC-1 human oral cancer cells with negligible toxic effects on the normal human hTRET-OME cells. The IC50 of Voacangine was 9 µM against SCC-1 cells relative to IC50 of 100 µM against normal hTRET-OME cells. The reduction of the proliferative rates was attributed to the induction of ROS triggered apoptosis which was associated with activation of Caspase-3, upregulation of Bax and suppression of Bcl-2. Voacangine induced G2/M cell cycle arrest in a dose-dependent manner. Additionally, the anticancer effects of Voacangine on oral cancer cells were exerted through the inhibition of PI3K/AKT signaling cascade. CONCLUSION: Taken all together, we conclude that Voacangine is a potent anticancer molecule and may be utilized for the development of systemic therapy for oral cancer.

Two novel metal-organic frameworks based on pyridyl-imidazole-carboxyl multifunctional ligand: selective CO2 capture and multiresponsive luminescence sensor.

Two novel metal-organic frameworks, formulated as [Mn(CIP-)2] (1) and [Ag(CIP-)] (2) (HCIP = 4-(4-carboxylphenyl)-2,6-di(4-imidazol-1-yl)phenyl)pyridine), were solvothermally synthesized based on a pyridyl-imidazole-carboxyl multifunctional ligand. The single-crystal X-ray diffraction analysis shows that complex 1 is a 3D microporous framework with uncoordinated imidazole groups, and complex 2 is a 2D + 2D → 2D 3-fold parallel interpenetrated network. Complex 1 exhibited excellent CO2 selective absorption over N2 and CH4. IAST calculations revealed that the selectivities of 1 for the CO2/CH4 (50 : 50) and CO2/N2 (15 : 85) mixtures were 8.0 and 117 at 273 K under 1 bar, respectively. Moreover, the luminescence investigations displayed that complex 2 is an excellent MOF-based multiresponsive fluorescent probe for Fe3+, CrO42-/Cr2O72- and the pesticide 2,6-Dich-4-nitroaniline, with high selectivity and sensitivity. Notably, complex 2 exhibited a highly sensitive sensing ability (5.2 × 104 M-1) and a low detection limit (1.7 × 10-7 M) for 2,6-Dich-4-nitroaniline. To the best of our knowledge, this is the first Ag-MOF-based fluorescent sensor that can simultaneously detect metal ions, inorganic anions and pesticides.

Scaffolds for epithelial tissue engineering customized in elastomeric molds.

Restoration of soft tissue defects remains a challenge for surgical reconstruction. In this study, we introduce a new approach to fabricate poly(d,l-lactic acid) (PDLLA) scaffolds with anatomical shapes customized to regenerate three-dimensional soft tissue defects. Highly concentrated polymer/salt mixtures were molded in flexible polyether molds. Microcomputed tomography showed that with this approach it was possible to produce scaffolds with clinically acceptable volume ratio maintenance (>90%). Moreover, this technique allowed us to customize the average pore size and pore interconnectivity of the scaffolds by using variations of salt particle size. In addition, this study demonstrated that with the increasing porosity and/or the decreasing of the average pore size of the PDLLA scaffolds, their mechanical properties decrease and they degrade more slowly. Cell culture results showed that PDLLA scaffolds with an average pore size of 100 µm enhance the viability and proliferation rates of human gingival epithelial cells up to 21 days. The simple method proposed in this article can be extended to fabricate porous scaffolds with customizable anatomical shapes and optimal pore structure for epithelial tissue engineering. © 2017 Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 880-890, 2018.

Suppression of lung cancer progression by isoliquiritigenin through its metabolite 2, 4, 2', 4'-Tetrahydroxychalcone.

BACKGROUND: Licorice is an herb extensively used for both culinary and medicinal purposes. Various constituents of licorice have been shown to exhibit anti-tumorigenic effect in diverse cancer types. However, majority of these studies focus on the aspect of their growth-suppressive role. In this study, we systematically analyzed known licorice's constituents on the goal of identifying component(s) that can effectively suppress both cell migration and growth. METHODS: Effect of licorice's constituents on cell growth was evaluated by MTT assay while cell migration was assessed by both wound-healing and Transwell assays. Cytoskeleton reorganization and focal adhesion assembly were visualized by immunofluorescence staining with labeled phalloidin and anti-paxillin antibody. Activity of Src in cells was judged by western blot using phosphor-Src416 antibody while Src kinase activity was measured using Promega Src kinase assay system. Anti-tumorigenic capabilities of isoliquiritigenin (ISL) and 2, 4, 2', 4'-Tetrahydroxychalcone (THC) were investigated using lung cancer xenograft model. RESULTS: Using a panel of lung cancer cell lines, ISL was identified as the only licorice's constituent capable of inhibiting both cell migration and growth. ISL-led inhibition in cell migration resulted from impaired cytoskeleton reorganization and focal adhesion assembly. Assessing the phosphorylation of 141 cytoskeleton dynamics-associated proteins revealed that ISL reduced the abundance of Tyr421-phosphorylation of cortactin, Tyr925- and Tyr861-phosphorylation of FAK, indicating the involvement of Src because these sites are known to be phosphorylated by Src. Enigmatically, ISL inhibited Src in cells while displayed no effect on Src activity in cell-free system. The discrepancy was explained by the observation that THC, one of the major ISL metabolite identified in lung cancer cells abrogated Src activity both in cells and cell-free system. Similar to ISL, THC deterred cell migration and abolished cytoskeleton reorganization/focal adhesion assembly. Furthermore, we showed both ISL and THC suppressed in vitro lung cancer cell invasion and in vivo tumor progression. CONCLUSION: Our study suggests that ISL inhibits lung cancer cell migration and tumorigenesis by interfering with Src through its metabolite THC. As licorice is safely used for culinary purposes, our study suggests that ISL or THC may be safely used as a Src inhibitor.

Overexpression of MARCKS indicates a poor prognosis of oral squamous cell carcinoma.

Myristoylated alanine-rich C kinase substrate (MARCKS) is a protein kinase C substrate functioning in different physiological and pathological mechanisms. Previous studies have suggested that MARCKS is capable of influencing tumorigenesis and progression. However, a limited number of studies are available regarding the role of MARCKS in oral squamous cell carcinoma (OSCC). The present study primarily examined MARCKS expression in the OSCC tissues. Furthermore, increased expression of MARCKS was confirmed in the majority of OSCC tissues. Increased MARCKS expression was correlated with more advanced tumor stages, lymphatic metastasis and a poorer overall patient survival. Further molecular mechanistic examinations revealed that downregulated MARCKS expression inhibited the proliferation and migration of OSCC cells in vitro through interruption of MARCKS expression. In addition, the present study demonstrated that MARCKS aggravated OSCC progression via the phosphoinositide 3-kinase/protein kinase B pathway. Accordingly, the present study considered MARCKS to be a promoter of OSCC tumorigenesis and progression, with the potential utility as a biomarker of a poor prognosis.