Effects of Engineered Nanoparticles on the Enantioselective Transformation of Metalaxyl Agent and Commercial Metalaxyl in Agricultural Soils

The adsorption coefficient of racemic metalaxyl onto an agriculture soil was small and nonenantioselective. Biotransformation was the predominant pathway for the elimination of <i>R</i>-metalaxyl, while abiotic and biotransformation made a comparable contribution to the degradation of <i>S</i>-metalaxyl. Metalaxyl acid was the main transformation intermediate. The enantiomer fraction of metalaxyl decreased with an increase in its initial spike concentration or the presence of the co-constituents in metalaxyl commercial products. Under simulated solar irradiation, the presence of TiO₂ promoted the overall transformation kinetics through enhanced biotransformation and extra photoinduced chemical reactions. The promotion was enantioselective and thereafter changed the enantiomer fraction. The results obtained in this study showed that some achiral parameters, although they have no direct impact on enantioselective reactions with enantiomers, can significantly affect the enantioselective transformation of racemic metalaxyl. Thus, our results indicate that the contribution of chemical interactions on the enantioselective transformation of chiral pesticides may be underestimated

DataSheet_1_DNAAF5 promotes hepatocellular carcinoma malignant progression by recruiting USP39 to improve PFKL protein stability.docx

PurposesDynein axonemal assembly factor 5 (DNAAF5) is the transcription factor of regulating the cytoskeleton and hydrodynamic protein complex assembly, however, it was not well elucidated in the malignant progression of hepatocellular carcinoma (HCC).MethodsWe investigated the role of DNAAF5 in hepatocellular carcinoma by using multiple groups of clinical tissues combined with data from the TCGA database. Then we overexpressed DNAAF5 in hepatocellular carcinoma tumor tissues, which correlates with poor patient survival outcomes. Furthermore, we constructed stable cell lines of HCC cells to confirm the cancer-promoting effects of DNAAF5 in hepatocellular carcinoma. To explore the mechanisms of DNAAF5, transcriptome sequencing combined with mass spectrometry was also performed, which showed that DNAAF5 affects its downstream signaling pathway by interacting with PFKL and that DNAAF5 regulates PFKL protein stability by recruiting the deubiquitination protein, USP39. To corroborate these findings, the same series of tissue microarrays were used to confirm correlations between DNAAF5 and PFKL expressions. In animal experiments, DNAAF5 also promoted the proliferation of HCC cells.ResultsWe found that DNAAF5 expressions were markedly higher in HCC tissues, compared to the adjacent normal tissues. Increased levels of DNAAF5 were associated with significantly worse prognostic outcomes for HCC patients. Cell function experiments showed that HCC cells of overexpressing DNAAF5 exhibited faster proliferation rates, stronger clone formation abilities and higher drug resistance rates. However, tumor cell proliferation rates and colony formation were significantly decreased after DNAAF5 knockout, accompanied by an increase in sensitivity to sorafenib. In addition, the results of our study showed that DNAAF5 accelerates PFKL protein deubiquitination by recruiting USP39 in HCC cells. Furthermore, The overexpression of DNAAF5 could promote HCC cell proliferation in vivo and in vitro, whereas USP39 knockdown inhibited this effect. Overall, DNAAF5 serves as a scaffold protein to recruit USP39 to form a ternary complex by directly binding the PFKL protein, thereby improving the stability of the latter, which promotes the malignant process of hepatocellular carcinoma.ConclusionsThese findings revealed DNAAF5 was negatively correlated with the prognosis of patients with hepatocellular carcinoma. It underlying mechanism showed that DNAAF5 directly binds PFKL and recruits the deubiquitinated protein (USP39) to improve the stability of the PFKL protein, thus enhancing abnormal glycolysis in HCC cells.</p

Additional file 2: of Long non-coding RNA MIAT promotes gastric cancer growth and metastasis through regulation of miR-141/DDX5 pathway

Figure S1. MIAT was up-regulated in GC cell lines. Figure S2. MIAT depletion inhibited GC cell proliferation by cell cycle arrest and apoptosis. Figure S3. MIAT deletion suppressed GC growth in vivo. Figure S4. MIAT deletion regulated miRNAs expression. Figure S5. MIAT and miR-141 regulated DDX5 expression. (DOC 2958 kb

ART suppressed cervical cancer metastasis in vivo.

<p>HeLa cells were injected into nude mice via the tail vein. Following injection, the mice (ten mice per group) were treated with 100 mg/kg ART or vehicle (sterile PBS) by subcutaneous injection once per day. After 6 weeks, (A) the numbers of tumor nodules on lung surfaces, (B) the weight of the lungs and (C) COX-2 expression was measured. * P < 0.05, **P < 0.01, compared to control.</p

ART inhibited HOTAIR expression in cervical cancer cells.

<p>(A) The expression of HOTAIR, CCAT2, MALAT1, EBIC and MEG3 was measured in metastatic nodules from mice treated with ART or PBS. (B) The HOTAIR expression was measured in CaSki and Hela cells treated with or without ART. **P < 0.01, compared to control. **P < 0.01, compared to control.</p

Downregulation of HOTAIR inhibits COX-2 expression and PGE₂ production in cervical cancer cells.

<p><b>(A)</b> CaSki and Hela cells were transfected with 3 HOTAIR-specific siRNAs (siHOTAIR1, siHOTAIR2 and siHOTAIR3) for 24 h, then, HOTAIR expression was measured. CaSki and Hela cells were transfected with siHOTAIR3 (si-HOTAIR) for 24 h, then, (B) COX-2 protein level and (C) PGE₂ production were measured. **P < 0.01, compared to siNC.</p

COX-2 is required for the effect of HOTAIR on cervical cancer cell migration and invasion.

<p>CaSki cells were transfected with si-HOTAIR and pcDNA3.1-COX-2 for 48 h, then (A) COX-2 protein was measured and (B and C) transwell assays were performed. **P < 0.01, compared to siNC. ##P < 0.01, compared to si-HOTAIR + pcDNA3.1. After transfected with pcDNA3.1 or pcDNA3.1-COX-2 for 24 h, CaSki cells were treated with ART (50 mmol/l) for 24 h. Then, (D and E) transwell assays were performed. **P < 0.01, compared to control. ##P < 0.01, compared to pcDNA3.1 + ART.</p

HOTAIR interacts with COX-2 in cervical cancer cells.

<p>(A) Biotin-labeled HOTAIR or antisense RNA was incubated with nucleoprotein from CaSki cell extracts, and the protein of COX-2 and EZH2 was assayed by Western blot. A non-specific protein (GAPDH) was used as the control. (B) RIP experiments were performed in CaSki cells using an COX-2 antibody or non-specific IgG, and specific primers were used to detect HOTAIR and GAPDH.</p

Overexpression of HOTAIR reversed the effect of ART on cervical cancer cell migration and invasion.

<p>(A) CaSki and Hela cells was transfected with pcDNA3.1 or pcDNA3.1-HOTAIR for 24 h, then, HOTAIR expression was measured. After transfected with pcDNA3.1 or pcDNA3.1-HOTAIR for 24 h, CaSki and Hela cells were treated with ART (50 mmol/l) for 24 h. Then, (B) transwell assays were performed and (C) PGE₂ concentration was measured. **P < 0.01, compared to pcDNA3.1. ##P < 0.01, compared to pcDNA3.1 + ART.</p