Gene commander in the trash heap: Transcriptional regulation and ubiquitination modification mediated by RNF6 in carcinogenesis.

RING finger protein 6 (RNF6), a RING finger protein, has been identified as a potential tumor promoter in several cancers. However, the exact mechanism of RNF6 in cancer remains elusive. As in various diseases, RNF6 may be involved in regulating cell growth, cell proliferation, invasion, cell cycle progression, apoptosis and cell adhesion through E3 ligase-mediated ubiquitination. Thus, the research on RNF6 is mainly focused on the ubiquitination of RNF6 in recent years. This article summarizes the role of RNF6 ubiquitination in various physiological and pathological mechanisms, such as Akt/mTOR signaling pathway, Wnt/ß-catenin pathway, RNF6/ERα/Bcl-xL axis, and provides knowledge and understanding for the treatment of diseases.

Validation of ESM1 Related to Ovarian Cancer and the Biological Function and Prognostic Significance.

Background: Ovarian cancer (OC), a serious gynecological malignant disease, remains an enormous challenge in early diagnosis and medical treatment. Based on the GEO and TCGA databases in R language, endothelial cell-specific molecule 1 (ESM1) was confirmed separately with the bioinformatic analysis tool. ESM1 has been demonstrated to be upregulated in multiple cancer types, but the oncogenic mechanism by which ESM1 promotes OC is still largely unknown. Methods: In this study, we used WGCNA and random survival forest variable screening to filter out ESM1 in OC differentially expressed genes (DEGs). Next, we confirmed the mRNA and protein levels of ESM1 in OC samples via PCR and IHC. The correlation between the ESM1 level and clinical data of OC patients was further confirmed, including FIGO stage, lymph node metastasis, and recurrence. The role of ESM1 in OC development was explored by several functional experiments in vivo and in vitro. Then, the molecular mechanisms of ESM1 were further elucidated by bioinformatic end experimental analysis. Results: ESM1 was significantly upregulated in OC and was positively correlated with PFS but negatively correlated with OS. ESM1 knockdown inhibited cell proliferation, apoptosis escape, the cell cycle, angiogenesis, migration and invasion in multiple experiments. Moreover, GSVA found that ESM1 was associated with the Akt pathway, and our results supported this prediction. Conclusion: ESM1 was closely correlated with OC development and progression, and it could be considered a novel biomarker and therapeutic target for OC patients.

Systemic Analysis of the DNA Replication Regulator MCM Complex in Ovarian Cancer and Its Prognostic Value.

Microliposome maintenance (MCM) 2, MCM3, MCM4, MCM5, MCM6, and MCM7 are DNA replication regulators and are involved in the progression of multiple cancer types, but their role in ovarian cancer is still unclear. The purpose of this study is to clarify the biological function and prognostic value of the MCM complex in ovarian cancer (OS) progression. We analyzed DNA alterations, mRNA and protein levels, protein structure, PPI network, functional enrichment, and prognostic value in OC based on the Oncomine, cBioPortal, TCGA, CPTAC, PDB, GeneMANIA, DAVID, KEGG, and GSCALite databases. The results indicated that the protein levels of these DNA replication regulators were increased significantly. Moreover, survival analysis showed a prognostic signature based on the MCM complex, which performed moderately well in terms of OS prognostic prediction. Additionally, protein structure, functional enrichment, and PPI network analyses indicated that the MCM complex synergistically promoted OC progression by accelerating DNA replication and the cell cycle. In conclusion, our study suggested that the MCM complex might be a potential target and prognostic marker for OC patients.

Propofol­induced miR­125a­5p inhibits the proliferation and metastasis of ovarian cancer by suppressing LIN28B.

Propofol, a commonly used intravenous anesthetic agent during surgery, has relatively widespread pharmacological actions. Previous studies have reported that propofol may act as an antitumor drug in several cancer types, such as pancreatic cancer, lung cancer and gastric cancer. However, the underlying mechanism in ovarian cancer remain unknown. Therefore, the present study investigated the pharmacological effect of propofol on microRNAs (miRNAs) in ovarian cancer treatment. Propofol (1, 5 or 10 µg/ml) was used to treat A2780 and SKOV3 ovarian cancer cells for 1, 2, 3, 4 or 5 days. The MTT assay was used to detect cell viability, while wound healing and Transwell assays were utilized to assess the invasive and migratory abilities. The bioinformatics prediction approach identified differentially expressed miRNAs (miRs) that were used in Gene Ontology, Gene Set Enrichment Analysis and Kyoto Encyclopedia of Genes and Genomes analyses. The expression levels of miR­125a­5p and lin­28 homolog B (LIN28B) were evaluated by reverse transcription­quantitative PCR (RT­qPCR). A luciferase assay was performed to identify the relationship between miR­125a­5p and LIN28B. Western blotting was conducted to measure the protein expression of LIN28B. It was demonstrated that propofol significantly upregulated miR­125a­5p to exert its antitumor activity. RT­qPCR results suggested that propofol could upregulate miR­125a­5p and LIN28B expression levels in ovarian cancer cell lines. Western blot analysis also indicated that propofol could enhance the expression of LIN28B in ovarian cancer cell lines. The luciferase assay identified that miR­125a­5p could directly inhibit the expression of LIN28B to suppress proliferation and metastasis in ovarian cancer. In conclusion, these results suggested that propofol inhibited ovarian cancer proliferation and metastasis by enhancing miR­125a­5p, which targets LIN28B.

Identification and validation of a prognostic index based on a metabolic-genomic landscape analysis of ovarian cancer.

PURPOSE: Tumour metabolism has become a novel factor targeted by personalised cancer drugs. This research evaluated the prognostic significance of metabolism-related genes (MRGs) in ovarian serous cystadenocarcinoma (OSC). METHODS: MRGs in 379 women surviving OSC were obtained using The Cancer Genome Atlas (TCGA) database. Then, several biomedical computational algorithms were employed to identify eight hub prognostic MRGs that were significantly relevant to OSC survival. These eight genes have important clinical significance and prognostic value in OSC. Subsequently, a prognostic index was constructed. Drug sensitivity analysis was used to screen the key genes in eight MRGs. Immunohistochemistry (IHC) staining confirmed the expression levels of key genes and their correlations with clinical parameters and prognosis for patients. RESULTS: A total of 701 differentially expressed MRGs were confirmed in women with OSC by the TCGA database. The random walking with restart (RWR) algorithm and the univariate Cox and lasso regression analyses indicated a prognostic signature based on eight MRGs (i.e., ENPP1, FH, CYP2E1, HPGDS, ADCY9, NDUFA5, ADH1B and PYGB), which performed moderately well in prognostic predictions. Drug sensitivity analysis indicated that PYGB played a key role in the progression of OSC. Also, IHC staining confirmed that PYGB has a close correlation with clinical parameters and poor prognosis in OSC. CONCLUSION: The results of the present study may help to establish a foundation for future research attempting to predict the prognosis of OSC patients and to characterise OSC metabolism.

Thiol-ene Click Derivatization for the Determination of Acrylamide in Potato Products by Capillary Electrophoresis with Capacitively Coupled Contactless Conductivity Detection.

The development of analytical methods for acrylamide formed during food processing is of great significance for food safety, but limited by its inherent characteristics, the analysis of acrylamide is a continuing challenge. In this study, an efficient derivatization strategy for acrylamide based on thiol-ene click reaction with cysteine as derivatization reagent was proposed, and the resulting derivative was then analyzed by capillary electrophoresis with capacitively coupled contactless conductivity detection (CE-C4D). After systematic investigation including catalyst dosage (0-20 mM), reaction temperature (30-90 °C) and time (1-60 min), and cysteine concentration (0.2-3.6 mM), acrylamide could be efficiently labeled by 2.0 mM cysteine at 70 °C for 10 min using 4 mM n-butylamine as catalyst. Application of 10 mM triethylamine as separation buffer, the labeled acrylamide was analyzed within 2.0 min, and the relative standard deviations of migration time and peak area were less than 0.84% and 5.6%, indicating good precision. The C4D signal of acrylamide derivative showed a good linear relationship with acrylamide concentration in the range of 7-200 µM with the correlation coefficient of 0.9991. The limit of detection and limit of quantification were calculated to be 0.16 µM and 0.52 µM, respectively. Assisted further by the QuEChERS (quick, easy, cheap, effective, rugged, and safe) sample pretreatment, the developed derivatization strategy and subsequent CE-C4D method were successfully applied for the determination of acrylamide in potato products.

Carboplatin prodrug conjugated Fe3O4 nanoparticles for magnetically targeted drug delivery in ovarian cancer cells.

Platinum (Pt)-based drugs including cisplatin, carboplatin and oxaliplatin have been widely used as first-line anticancer regimens due to their excellent anticancer efficacy. However, the clinical application of these drugs is greatly limited due to their side effects and drug resistance. In this study, an antitumor drug delivery system based on carboplatin prodrug loading Fe3O4 nanoparticles (NPs@carboplatin) was developed and the antitumor activity was also investigated. The as-synthesized NPs@carboplatin has an average diameter of 7.88 nm with a zeta potential of 8.11 mV. It exhibited a higher cytotoxic effect than carboplatin on both A2780 (cisplatin sensitive) and A2780DDP (cisplatin resistant) ovarian cancer cells via MTT assay, which can overcome Pt resistance. Moreover, the nanoparticles (NPs) loaded with carboplatin possess excellent delivery capability, which can be effectively taken up by ovarian cancer cell lines through an endocytosis process. With excellent delivery capability as carriers, NPs@carboplatin can promote drug internalization and result in the increase of intracellular drug accumulation with enhanced cellular cytotoxicity. Furthermore, in vivo experiments demonstrated that NPs@carboplatin can be widely distributed into major organs, and in the presence of an external magnetic field, the Fe3O4 nanocarrier is beneficial to visualize the tumor site location and promote the subsequent antitumor efficacy. It revealed a relatively high tumor inhibition rate without obvious potential toxicity to normal organs. Taken together, these results indicate that the magnetically targeted NPs@carboplatin can act as a drug delivery system and will have great potential in ovarian cancer therapeutic applications.

Sensitive analysis of curcuminoids via micellar electrokinetic chromatography with laser-induced native fluorescence detection and mixed micelles-induced fluorescence synergism.

Curcuminoids, the major bioactive constituents of traditional medicine known as turmeric, have exhibited extensive therapeutic benefits. Excited by violet-blue light, curcuminoids can emit native fluorescence, making them possible to be detected with high sensitivity and specificity by laser-induced native fluorescence (LINF). Here, a commercial 445 nm laser diode was used as an excitation source to construct a confocal laser-induced fluorescence (LIF) detector and then a complete capillary electrophoresis (CE) system coupled with LIF detection was established. With three major curcuminoids, curcumin, demethoxy curcumin (DMC) and bisdemethoxy curcumin (BDMC) as target analytes, a micellar electrokinetic chromatography (MEKC) method was proposed using mixed micelles consisting of Triton X-100 and SDS to sensitize the native fluorescence of curcuminoids and enhance their separation efficiency. Fluorescence spectra revealed that the mixed micelles induced fluorescence synergism could enhance the signals of three curcuminoids by 77-, 57-, and 47-fold for curcumin, DMC, and BDMC. After systematic investigation, the optimal separation buffer for curcuminoids was chosen as follows: 20 mM Triton X-100, 20 mM SDS, 30% (v/v) methanol in 10 mM borax solution at pH 10.0. Under these conditions, a baseline separation of three curcuminoids was achieved within 10 min and the detection limits were found to be 4.1, 2.6, and 0.4 ng/mL for curcumin, DMC, and BDMC, respectively. Furthermore, the developed MEKC-LINF method was validated in terms of precision, linearity, accuracy and successfully applied for the determination of three curcuminoids in turmeric, medicinal turmeric liniment, curry seasoning, and human urine samples.