Application of Metabolomics in Fungal Research.

Metabolomics is an essential method to study the dynamic changes of metabolic networks and products using modern analytical techniques, as well as reveal the life phenomena and their inherent laws. Currently, more and more attention has been paid to the development of metabolic histochemistry in the fungus field. This paper reviews the application of metabolomics in fungal research from five aspects: identification, response to stress, metabolite discovery, metabolism engineering, and fungal interactions with plants.

Three new C21 steroidal glycosides isolated from Metaplexis japonica and their potential inhibitory effects on tyrosine protein kinases.

Three new steroidal glycosides, metapregnoside A-C (II-IV), together with one known compound, byzantionoside B (I), were isolated from the fresh whole herb of Metaplexis japonica by using high-speed countercurrent chromatography and semi-preparative liquid chromatography. Their structures and relative configurations were elucidated by spectroscopic methods including 1D NMR, 2D NMR and HR-ESI-MS. The potential targets of compound I-IV were identified by virtual screening. And the potential inhibitory effects of these compounds on tyrosine protein kinases were compared by molecular docking. Byzantionoside B (I) was the first isolated compound from Metaplexis genus. The docking score of metapregnoside C (IV) was the highest. And the sugar chain residues at position C-20 in the pregn-4-en-3-one derivatives is the main factor affecting their docking scores on tyrosine protein kinases Fes/Fps.

Immunotherapy Summary for Cytokine Storm in COVID-19.

COVID-19 pneumonia caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) has ravaged the world, resulting in an alarming number of infections and deaths, and the number continues to increase. The pathogenesis caused by the novel coronavirus was found to be a disruption of the pro-inflammatory/anti-inflammatory response. Due to the lack of effective treatments, different strategies and treatment methods are still being researched, with the use of vaccines to make the body immune becoming the most effective means of prevention. Antiviral drugs and respiratory support are often used clinically as needed, but are not yet sufficient to alleviate the cytokine storm (CS) and systemic inflammatory response syndrome. How to neutralize the cytokine storm and inhibit excessive immune cell activation becomes the key to treating neocoronavirus pneumonia. Immunotherapy through the application of hormones and monoclonal antibodies can alleviate the immune imbalance, but the clinical effectiveness and side effects remain controversial. This article reviews the pathogenesis of neocoronavirus pneumonia and discusses the immunomodulatory therapies currently applied to COVID-19. We aim to give some conceptual thought to the prevention and immunotherapy of neocoronavirus pneumonia.

A rapid and practical prediction method for the Arizona solvent system family used in high speed countercurrent chromatography.

Selecting the appropriate solvent system is the key to the successful separation of samples by using countercurrent chromatography. Although high-speed countercurrent chromatography has been widely used in the separation and preparation of natural products, the selection of a solvent system has always been a stumbling block to the application of high-speed countercurrent chromatography. In order to explore a rapid and practical prediction method to select countercurrent chromatography solvent system, five linear prediction models of the Arizona solvent system family (HEMW) was established by using fourteen compounds with different structures and five HPLC columns of different brands. And two different solvent system selection methods (The partition coefficient K of the target compound in the solvent system was in the range of 0.25 < K < 2.5) were proposed for targeted separation of compounds and multi-component separation in a complex sample respectively. The appropriate HSCCC solvent system of five known compounds was determined by a HPLC analysis and a shake flask test and the appropriate HSCCC solvent system of two Chinese herbal extracts was determined by a HPLC analysis to verify the prediction method. In this study, solid-liquid partition chromatography (HPLC) and liquid-liquid partition chromatography (HSCCC) were linked by polarity to simplify the screening process of solvent system. This method reduced the difficulty and workload of solvent system selection, which provided methods and ideas for more solvent system prediction models.

Secretory IgA in Mucosa of Pharynx and Larynx Plays an Important Role against Influenza A Virus Infection in Kidney Yang Deficiency Syndrome Model.

OBJECTIVE: Influenza virus poses a major threat to human health and has serious morbidity and mortality which commonly occurs in high-risk populations. Pharynx and larynx of the upper respiratory tract mucosa is the first defense line against influenza virus infection. However, the ability of the pharynx and larynx organ to eliminate the influenza pathogen is still not clear under different host conditions. METHODS: In this study, a mouse model of kidney yang deficiency syndrome (KYDS) was used to mimic high-risk peoples. Two different methods of influenza A (H1N1) virus infection by nasal dropping or tracheal intubation were applied to these mice, which were divided into four groups: normal intubation (NI) group, normal nasal dropping (ND) group, model intubation (MI) group, and model nasal dropping (MD) group. The normal control (NC) group was used as a negative control. Body weight, rectal temperature, and survival rate were observed every day. Histopathologic changes, visceral index, gene expressions of H1N1, cytokine expressions, secretory IgA (SIgA) antibodies of tracheal lavage fluids in the upper respiratory tract, and bronchoalveolar lavage fluids were analyzed by ELISA. RESULTS: The MD group had an earlier serious morbidity and mortality than the others. MI and NI groups became severe only in the 6th to 7th day after infection. The index of the lung increased significantly in NI, MI, and MD groups. Conversely, indices of the thymus and spleen increased significantly in NC and ND groups. H&E staining showed severe tissue lesions in MD, MI, and NI groups. H1N1 gene expressions were higher in the MD group compared with the MI group on the 3rd day; however, the MD group decreased significantly on the 7th day. IL-6 levels increased remarkably, and SIgA expressions decreased significantly in the MD group compared with the NC group. CONCLUSIONS: SIgA secretions are influenced directly by different conditions of the host in the pharynx and larynx in the upper respiratory tract mucosa. In the KYDS virus disease mode, SIgA expressions could be inhibited severely, which leads to serious morbidity and mortality after influenza A virus infection. The SIgA expressions of the pharynx and larynx would be an important target in high-risk populations against the influenza A virus for vaccine or antiviral drugs research.

A facile and sensitive electrochemical sensor for non-enzymatic glucose detection based on three-dimensional flexible polyurethane sponge decorated with nickel hydroxide.

A novel three-dimensional nickel hydroxide/polyurethane (Ni(OH)2/PU) electrode was prepared by a simple and environmentally friendly method and used for non-enzymatic detection of glucose. The Ni(OH)2/PU electrode was obtained by one-pot hydrothermal method of loading nickel hydroxide on a cheap, easily available and flexible polyurethane sponge, which is facile and energy-saving. The porous structure of the polyurethane sponge provides a large surface area and a rich electrochemical active site for the electrode, which is beneficial to the oxidation reaction of glucose on the surface of the electrode with Ni(OH)2. The Ni(OH)2/PU electrode structure was characterized by scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS). The cyclic voltammetry test was used to study the catalytic performance of Ni(OH)2/PU electrode for oxidation of glucose and the chronoamperometry was used to investigate the detection performance of Ni(OH)2/PU electrode on glucose. The results indicate that this non-enzymatic glucose sensor had a high sensitivity of 2845 µA mM-1 cm-2, a low detection limit of 0.32 µM (S/N = 3), a detection range of 0.01-2.06 mM and response time of less than 5 s. In addition, the Ni(OH)2/PU electrode had excellent selectivity, reproducibility and stability and also exhibited effective detection of glucose in fetal bovine serum (FBS). In summary, Ni(OH)2/PU electrode had broad prospects as an excellent candidate for non-enzymatic glucose sensors. The study also opens up a facile and energy-saving approach for preparing three-dimensional (3D) functionalized polymer electrode via hydrothermal method as electrochemical sensors.

Rational Design of Novel Efficient Palladium Electrode Embellished 3D Hierarchical Graphene/Polyimide Foam for Hydrogen Peroxide Electroreduction.

The electrocatalytic applications of traditional polyimide film and carbon nanomaterials are hindered due to a shortage of three-dimensional hierarchical conductivity and porous structure. Herein, a novel polyimide-based electrode based on a highly efficient palladium nanocatalyst embellished three-dimensional reduced graphene oxide/polyimide foam (Pd/3D RGO@PI foam, signed PRP) toward H2O2 electroreduction was designed and prepared through thermal foaming procedure, followed by facile dip-drying method and electrodeposition. As expected, such a binder-free, 3D hierarchical structure PRP electrode presented high catalytic property, good stability, as well as low activation energy toward H2O2 electroreduction during the electrochemical measurement period. The PRP electrode showed a reduction current density of 810 mA·cm-2 at -0.2 V (vs Ag/AgCl) in 2.0 mol·L-1 H2SO4 and 2.0 mol·L-1 H2O2. Moreover, the PRP electrode also illustrated good reproducibility and repeatability. Reproducibility presented almost 95.8% of the initial current density after 1000 cycles test. Also, the activation energy of H2O2 electroreduction on 3D PRP electrode was 21.624 kJ·mol-1. Benefiting from the 3D hierarchical structure and efficient catalyst, the PRP electrode exhibited excellent electrocatalytic performance and was considered to be a potential candidate material for fuel cells.

Isolation, purification, structural analysis and biological activities of water-soluble polysaccharide from Glehniae radix.

A new polysaccharide named GRP (Glehniae radix polysaccharide) was isolated and purified from Glehniae radix by hot water extraction, ethanol precipitation, anion-exchange and gel-filtration chromatography. GRP was homogeneous, with a molecular weight of 1.33×104Da, as determined by high-performance size-exclusion chromatography-refractive index detector analysis. Its structural characteristics were investigated and elucidated by methylation analysis, gas chromatography mass spectrometry, Fourier transform infrared and nuclear magnetic resonance spectroscopy. Based on obtained data, GRP was found to be α­d­glucan containing (1→6)-linked and (1→3)-linked backbone with a branch of one (1→6)-linked and terminal glucoses submitting at the C-4 position every fourteen residues. The biological activities of GRP upon proliferation of splenic lymphocyte, RAW264.7 cells and A549 cell, and production of nitric oxide (NO) were investigated in vitro. The results showed that GRP exhibited inhibition against A549 cells proliferation and NO production in RAW264.7 cells, and displayed promotion for proliferation of mouse spleen lymphocytes and RAW264.7 cells, which suggested that GRP may have potential immunoregulation, anti-inflammatory and anti-tumor activity.

Simultaneous quantification of fifteen compounds in rat plasma by LC-MS/MS and its application to a pharmacokinetic study of Chaihu-Guizhi decoction.

A simple, sensitive and selective high-performance liquid chromatography electrospray ionization tandem mass spectrometry (HPLC-ESI-MS/MS) method was developed and validated for simultaneous determination and pharmacokinetic study of 15 active compounds (Saikosaponin A, Baicalin, Wogonin, Glycyrrhizic acid, Glycyrrhetinic acid, Albiflorin, Paeoniflorin, Liquiritin, Isoliquiritin, Liquiritigenin, Isoliquiritigenin, Cinnamic acid, Gallic acid, Wogonoside and Oroxylin A) in rat plasma. After a feasible protein precipitation using methanol for sample preparation, chromatographic separation was carried out with a Halo® C18 column (2.1 × 100 mm, 2.7 µm) at 35 °C, water containing 0.1% formic acid and acetonitrile were used as the mobile phase with a flow rate of 0.3 mL/min. Multiple reaction monitoring (MRM) with positive and negative ion switching mode was performed for the quantification of the standards and internal standard in plasma. All the calibration curves showed good linear regression within the linear range (r2 > 0.9923). In particular, the results of the method validation including specificity, linearity, accuracy, precision, extraction recovery, matrix effect, and stability of compounds in bio-samples were all within the current acceptance criteria. The established method was successfully applied to the pharmacokinetic study of 15 compounds in rats after oral administration of CGD and laid the foundation for studying the active components and mechanism of CGD in vivo.

Proteomic analysis reveals that pheophorbide a-mediated photodynamic treatment inhibits prostate cancer growth by hampering GDP-GTP exchange of ras-family proteins.

BACKGROUND: We previously reported that pheophorbide a (PhA), excited by 630 nm light, significantly inhibited the growth of prostate cancer cells. In this study, we employed whole-cell proteomics to investigate photodynamic treatment (PDT)-related proteins. METHODS: Two-dimensional gel electrophoresis (2-DE) coupled with tandem mass spectrometry was employed to reveal the proteins involved in PhA-mediated PDT in LNCaP and PC-3 prostate cancer cells. RESULTS: After PhA-PDT treatment, decreased expression of translationally-controlled tumor protein (TCTP) was found in both PC-3 and LNCaP whole-cell proteomes. In contrast, human rab GDP dissociation inhibitor (GDI) in LNCaP cells and ras-related homologs GDI in PC-3 cells were up-regulated. CONCLUSIONS: GDP-GTP exchange is an underlying target of photodynamic treatment in prostate cancer cells.

[Action mechanism of Mahuang Xixin Fuzi decoction for mice with influenza based on metabolomics information].

This study aimed to analyze the endogenous metabolite changes in the serum of mice infected with H1N1 virus after intervention by Mahuang-Xixin-Fuzi decoction (MXF) based on metabolomics method, investigate potential biomarkers and related metabolic pathways, and explore the therapeutic mechanism of MXF through metabolomics technology. Thirty-six Kunming (KM) mice were randomly divided into three groups: normal group, model group and MXF group. Influenza virus H1N1 was used by nasal drip to establish influenza mice model. The mice in MXF group were orally administrated with MXF for 6 consecutive days after inoculation, and the other two groups were given with equal volume of saline solution in the same way. Body weight, rectal temperature, morbidity and mortality were recorded daily. Serum samples were collected 24 hours after the last administration for HPLC-TOF-MS analysis. The results showed that as compared with the normal group, the body weight and rectal temperature were decreased in model group, and their lung index and mortality rate were significantly increased (P<0.05); MXF had good therapeutic effects on the abnormity of body weight, rectal temperature, lung index and high mortality rate of mice infected with H1N1 virus. The original data collected from the serum samples were analyzed with R language, MPP, SIMCA-P and other software, and significant changes were found in 14 kinds of endogenous substances from mice serum (P<0.05). As compared with model group, the potential metabolic markers in MXF group recovered to normal levels to a certain degree after being intervened by MXF. Further analysis with MetPA data platform showed that, the pathways involved in 14 metabolites included glucose metabolism, arachidonic acid metabolism, glycerophospholipids and sphingolipids metabolism etc. The metabolomics study and pharmacological experiment showed that MXF might play a role of efficacy by improving glucose metabolism, regulating arachidonic acid metabolism, glycerophospholipid and sphingolipid metabolic pathways.

Grifolin induces apoptosis and promotes cell cycle arrest in the A2780 human ovarian cancer cell line via inactivation of the ERK1/2 and Akt pathways.

Grifolin, a secondary metabolic product isolated from the mushroom Albatrellus confluence, has been demonstrated to possess antitumor activities in a variety of malignant cells. However, the signaling pathways and the molecular mechanisms underlying the anticancer effects of the agent in human ovarian cancer remain to be elucidated. The aim of the present study was to examine the effect of grifolin treatment on the human ovarian cancer cell line, A2780. MTT and flow cytometry analysis were used to analyze the viability of A2780 cells following treatment with grifolin. Western blotting was used analyze the expression of apoptosis-associated and cell cycle arrest-associated proteins. The results of MTT assays and flow cytometry analysis revealed that grifolin suppressed cell viability, induced apoptosis and triggered cell cycle arrest. Western blotting revealed that grifolin treatment resulted in inactivation of protein kinase B (Akt) and extracellular signal-related kinase 1/2 (ERK1/2), accompanied by upregulation of Bcl-2 associated X, apoptosis regulator, cleaved-caspase-3 and cleaved-poly (ADP-ribose) polymerase, and downregulation of B cell lymphoma-2, cyclin dependent kinase 4 and cyclinD1. The results of the present study indicated that grifolin had significant anti-cancer effects on the human ovarian cancer A2780 cells, which occurred via the Akt and ERK1/2 signaling pathways to at least a certain extent. These results demonstrate the therapeutic potential of grifolin as a treatment for ovarian cancer.

Forkhead transcription factor 1 inhibits endometrial cancer cell proliferation via sterol regulatory element-binding protein 1.

The morbidity and mortality associated with endometrial cancer (EC) has increased in recent years. Regarded as a tumor suppressor, forkhead transcription factor 1 (FOXO1) has various biological activities and participates in cell cycle progression, apoptosis and differentiation. Notably, FOXO1 also functions in the regulation of lipogenesis and energy metabolism. Lipogenesis is a feature of cancer and is upregulated in EC. Sterol regulatory element-binding protein 1 (SREBP1) is a transcription factor that is also able to regulate lipogenesis. Increased expression of SREBP1 is directly correlated with malignant transformation of tumors. A previous study demonstrated that SREBP1 was highly expressed in EC and directly resulted in tumorigenesis. However, the association between FOXO1 and SREBP1 in EC is not clear. In the present study, lentiviruses overexpressing FOXO1 were used in cell transfection and transduction. Cell viability assays demonstrated that the overexpression of FOXO1 was able to suppress cell proliferation significantly in Ishikawa and AN3 CA cell lines. In addition, FOXO1 overexpression significantly inhibited cell migration and invasion ability in vitro. In xenograft models, overexpression of FOXO1 suppressed cell tumorigenesis, and western blot analysis demonstrated that SREBP1 expression was markedly reduced in the FOXO1-overexpressing cells. It may therefore be concluded that FOXO1 is able to inhibit the proliferative capacity of cells in vitro and in vivo, in addition to the migratory and invasive capacities in vitro by directly targeting SREBP1.

Grifolin induces autophagic cell death by inhibiting the Akt/mTOR/S6K pathway in human ovarian cancer cells.

Grifolin, a secondary metabolic product isolated from the mushroom Albatrellus confluence, has been reported to possess antitumor activities in various tumors. To date, no report exists on the role of autophagy in grifolin-treated human ovarian cancer cells. In the present study, we investigated the effect and the mechanism of autophagy in ovarian cancer. Ovarian cancer cell lines A2780 and SKOV3 were treated with grifolin. Cell proliferation was assessed by MTT assay and the autophagic effect was determined using flow cytometry, electron microscopy, immunofluorescence staining and GFP-LC3 puncta formation assay. The expression of autophagy markers and the main autophagy-associated Akt/mTOR/S6K pathway proteins were measured by western blot analysis. MTT assay indicated that grifolin inhibits the proliferation of human ovarian cancer cell lines A2780 and SKOV3. Flow cytometry, electron microscopy, immunofluorescence and GFP-LC3 puncta formation assay proved that grifolin induces autophagic cell death in human ovarian cancer. The results of the western blot analysis suggested that grifolin treatment leads to upregulation of autophagy markers LC3B, Atg7, Beclin-1 along with downregulation of P62. In addition, the proteins of the pathways p-Akt, p-mTOR, p-p70S6K and p-4E-BP1 were downregulated while the total of these proteins remained unaffected. The present study indicated that grifolin could induce autophagic cell death in human ovarian cancer by inhibiting the Akt/mTOR/S6K pathway.

[The Component Analysis of Petroleum Ether Extract in Scutellariae barbatae,Hedyotis diffusa and the Herb Pair and the Investigation of Anti-Endometrial Cancer Cells Activity].

Objective: To investigate the chemical components and the activity of anti-endometrial cancer cells of the petroleum ether extract in Scutellariae barbatae and Hedyotis diffusa and the herb pair. Methods: Main composition analysis and identification were determined by the GC-MS technology combined with Kovats retention index( KI). Activity of anti-endometrial cancer cells was researched by MTT assay. Results: Unsaturated fatty acid,esters,sterol and other compounds in Scutellariae barbatae,Hedyotis diffusa and the herb pair were identified by GC-MS. Hedyotis diffusa and the herb pair contained more anthraquinones which distinguished from Scutellariae barbatae. The IC50 values for HEC-1A cells of petroleum ether extract in Scutellariae barbatae and Hedyotis diffusa and the herb pair were 275. 204 µg / m L,105. 826 µg / m L,148. 645 µg / m L. The IC50 values for Ishikawa cells of petroleum ether extract in Scutellariae barbatae and Hedyotis diffusa and the herb pair are 189. 114 µg / m L,77. 974 µg / m L,137. 999 µg / m L. Conclusion: Petroleum ether extract in Scutellariac barbatae and Hedyotis diffusa and the herb pair has inhibition effect on the proliferation of HEC-1A and Ishikawa cells,the Hedyotis diffusa has the strongest activity of anti-endometrial cancer. It is speculated that the strongest activity could be related to the higher content of anthraquinones.

Biglycan enhances the ability of migration and invasion in endometrial cancer.

OBJECTIVE: This study aimed to confirm that biglycan (BGN) can promote the migration and invasion in endometrial cancer both in vitro and in vivo and the possible therapeutic value of BGN in endometrial cancer. METHODS: Western blot was used to screen out the higher protein level of BGN in human endometrial cancer cells; BGN knocked down cells were constructed by lentiviral transfection; The effect of BGN in endometrial cancer detected by wound healing, transwell migration, and invasion, endothelial tube formation assay in vitro, and xenograft model in vivo. RESULTS: (1) We found that BGN expression level is higher in the Ishikawa (ISK, high differentiation) and AN3CA (poor differentiation) cells than other endometrial cancer cells. (2) BGN enhances endometrial cancer cell wound healing, invasion, and migration ability and formation ability of endothelial cells in vitro. Xenograft model has confirmed the outcome in vivo. CONCLUSIONS: BGN might play an important role on metastasis in human endometrial cancer and it might be a target marker for the molecular therapy of advanced and recurrence endometrial cancer.

SIRT1 promotes endometrial tumor growth by targeting SREBP1 and lipogenesis.

Silent information regulator 1 (SIRT1) is involved in a number of cellular regulatory mechanisms affecting cellular life span, stress resistance, apoptosis and cellular metabolism. Recent studies have revealed that SIRT1 plays a dual role as a tumor suppressor and a tumor promoter in multiple stages of carcinogenesis. Increased lipogenesis has been found in cancer cells, sterol regulatory element binding protein 1 (SREBP1) are nuclear lipogenic transcription factors, which mainly regulate lipogenic processes by activating genes involved in fatty acid and triglyceride biosynthesis. In the present study, we detected expression of SIRT1 in endometrial cancer (EC) and illustrated the relationship between SIRT1 and SREBP1, which indicated that SIRT1 could stimulate endometrial tumor growth through the lipogenic pathway. Gene expression levels of SIRT1 were assayed using quantitative real-time PCR and protein expression levels were detected by western blotting. RNA interference was conducted in order to explore the subsequent effect on tumor cells and on the expression of SREBP1. Expression levels of SIRT1 in EC were found to be significantly higher than in normal endometrium. Knockdown of SIRT1 could downregulate expression of SREBP1 and suppress cell proliferation. These results demonstrated that SIRT1 may play a role as a tumor promoter in EC and can promote endometrial tumor growth by promoting lipogenesis. Our findings suggest that targeting SIRT1 may provide a theoretical basis for the management of EC.

[GC-MS analysis for volatile components from alpiniae katsumadai semen by three extraction methods].

OBJECTIVE: To analysis the volatile components in Alpiniae Katsumadai Semen. METHODS: The volatile components were extracted from Alpiniae Katsumadai Semen by steam distillation, head space injection and supercritical fluid extraction respectively, and then analyzed by GC-MS combined with Kovat's retention index. RESULTS: The volatile components extracted by steam distillation or head space extraction were found more likely to be terpenoids, whereas components extracted by supercritical fluid extraction were more likely to be alkenes, alcohols and aromatic compounds. CONCLUSION: Different sample pre-treatment methods are focused on different types of volatile components; Identification of the volatile components by GC-MS combined with Kovat's retention index is more accurate and rapid.

Single nucleotide polymorphism of SREBF-1 gene associated with an increased risk of endometrial cancer in Chinese women.

AIM: Elevated levels of sterol regulatory element-binding protein-1 (SREBP-1) have been found in endometrial cancer (EC), suggesting that it is essential to the development of EC. Obesity and diabetes have been established as known risk factors of EC, while SREBF-1 gene polymorphisms have also been found to be associated with obesity and type II diabetes. Therefore, we hypothesize that single nucleotide polymorphism (SNP) in SREBF-1 gene may be associated with increased risk of EC. METHOD: We analyzed the sequence of SREBF-1 in tissue samples from 30 EC cases and 6 benign controls using high throughput method. Based on the primary results, we selected one SNP (rs2297508) as a genetic marker to conduct a hospital-based case-control study with 139 EC cases and 129 benign controls. The samples were examined under the microscope to determine their histopathology prior to the SNP analysis using RT-PCR. RESULTS: Through sequence analysis, we found 10 SNPs of SREBF-1 associated with EC, including 3 new SNPs. Fourteen percent of EC showed the rs2297508 SNP with C allele, while only 7% had the C allele was present in benign controls (p = 0.027, OR = 1.983). Additionally, the C allele was associated with cancer differentiation (p<0.05) and the depth of myometrial invasion (p<0.05). CONCLUSION: Our study indicates that SNP (rs2297508) of SREBF-1 may serve as a genetic predisposition factor for the development of EC and screening of such genetic marker may be helpful in its early detection.

Sterol regulatory element-binding protein-1/fatty acid synthase involvement in proliferation inhibition and apoptosis promotion induced by progesterone in endometrial cancer.

BACKGROUND: The number of endometrial cancer (EC) cases is escalating rapidly, with no evident improvements in survival rates. The downregulation of progesterone receptor, resulting in progestin resistance, is presently a major problem regarding the therapeutic aspect. On the basis of this, we can focus more on the downstream signaling pathways that are controlled by progesterone. Lipid biosynthesis mediated by sterol regulatory element-binding protein-1/fatty acid synthase (SREBP-1/FASN) is of utmost importance to the growth and the proliferation of EC cells, so we hypothesize that SREBP-1/FASN might be involved in suppressing the proliferation and promoting apoptosis in EC cells through the effects induced by progesterone. MATERIAL AND METHODS: The Cell Counting Kit-8 was used to analyze the growth inhibition ratio of Ishikawa cells upon treatment with megestrol acetate (MA; MA is a progesterone derivative, also known as 17α-acetoxy-6-dehydro-6-methylprogesterone) and to determine the 50% inhibitory concentration. Apoptosis ratio was analyzed by treatment of the cells with MA at 50% inhibitory concentration at different time intervals using Annexin V-FITC/propidium iodide. The protein and messenger RNA levels of SREBP-1 and FASN were compared between the experimental and control groups (MA-treated Ishikawa cells were considered to be the experimental group). RESULTS: The experimental group showed obvious growth inhibition that was time and concentration dependent. The apoptosis ratio was also significantly higher in the experimental group compared with the control group (P < 0.01). The protein and messenger RNA levels of SREBP-1 and FASN were significantly reduced by MA too. CONCLUSIONS: Sterol regulatory element-binding protein-1/FASN is involved in the proliferation suppression and apoptosis promotion brought about by MA in Ishikawa cells.