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Front Genet ; 11: 847, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32973867


Introduction: WD repeat domain phosphoinositide-interacting protein 3 (WIPI3) is a member of the WIPI protein family, autophagy marker, that is associated with the malignant progression of various human cancers, but its role in hepatocellular carcinoma (HCC) is still unclear. Materials and Methods: Firstly, we collected the mRNA expression of WIPI3 in HCC through the platform of Oncomine, as well as the DNA copy number variations (CNVs), and verified it on human HCC cell line and the GEO database. Then, the subgroups and prognosis of HCC were performed by the UALCAN web tool. The mutation of WIPI3 was analyzed by cBioPortal. The coexpression of WIPI3 in HCC was identified from the LinkedOmics database, and function enrichment analysis was done using the LinkFinder module in LinkedOmics. Coexpression gene network was constructed through the STRING database, and the MCODE plug-in of which was used to build the gene modules; both of them were visualized by the Cytoscape software. Finally, the top modular genes in the same patient cohort were constructed through data mining in The Cancer Genome Atlas (TCGA) liver hepatocellular carcinoma (LIHC) by using the UCSC Xena browser. Results: The results indicated that WIPI3 was frequently overexpressed in HCC, which could lead to a poor prognosis through the Kaplan-Meier (KM) analysis. Moreover, there existed mutations of WIPI3 in HCC, and the prognosis of WIPI3-altered group was significantly poor based on KM plotter data. Coexpression analysis showed that the coexpression gene of WIPI3 was associated with cell cycle and spliceosome. Further analysis suggested that WIPI3 and eukaryotic translation initiation factor 4A3 (EIF4A3) coordinately regulated the cancer cell cycle by spliceosome as a result of the strong positive correlation between them. Conclusion: In summary, WIPI3 is constantly overexpressed in HCC tissues, resulting in a poor prognosis; therefore, we can identify it as an effective target for the treatment of HCC.

Nanoscale ; 11(6): 2624-2630, 2019 Feb 07.
Artigo em Inglês | MEDLINE | ID: mdl-30693354


The great challenge in preparing a biomimetic enzyme sensor is to have sensitivity and selectivity equal to or better than its corresponding biological sensor. Porous electrodes possess a large surface area and are often used to greatly improve the sensor sensitivity. However, how to tailor the pore structure, especially the pore size distribution to further improve the sensitivity and selectivity of a biomimetic sensor, has not been investigated yet. The superoxide anion (O2˙-) plays essential roles in various biological processes and is of importance in clinical diagnosis and life science research. It is generally detected by the superoxide dismutase enzyme. Herein, we delicately tailor the pore structure of carbon nanofibers (CNFs) by pyrolysis to obtain an optimal mesopore structure for strong adsorption of DNA, followed by guiding the growth of Mn3(PO4)2 as a biomimetic enzyme toward highly sensitive detection of O2˙-. The Mn3(PO4)2-DNA/CNF sensor achieves the best sensitivity among the reported O2˙- sensors while possessing good selectivity. The enhancement mechanism is also investigated, indicating that the mesopore ratio of CNFs plays an essential role in the high sensitivity and selectivity due to their strong adsorption of DNA for guiding the growth of a large amount of uniform sensing components, Mn3(PO4)2, toward high sensitivity and selectivity. The biomimetic sensor was further used to in situ monitor O2˙- released from human keratinocyte cells and human malignant melanoma cells under drug stimulation, showing high sensitivity to real-time quantitative detection of O2˙-. This work provides a highly sensitive in situ real-time biomimetic O2˙- sensor for applications in biological research and diagnosis, while shedding light on the enhancement mechanism of the pore structure, especially the pore size distribution of a porous electrode for high performance sensing processes.

Técnicas Biossensoriais/métodos , DNA , Enzimas , Nanotubos de Carbono/química , Compostos Organometálicos , Superóxidos/análise , Adsorção , Linhagem Celular Tumoral , DNA/química , DNA/metabolismo , Enzimas/química , Enzimas/metabolismo , Humanos , Nanofibras/química , Compostos Organometálicos/química , Compostos Organometálicos/metabolismo , Porosidade
Nan Fang Yi Ke Da Xue Xue Bao ; 37(12): 1609-1614, 2017 Dec 20.
Artigo em Chinês | MEDLINE | ID: mdl-29292253


OBJECTIVE: To construct a cell model of 4.1R gene knockout in murine macrophage cell line RAW264.7 using CRISPR/Cas9 technique. METHODS: Three high?grade small?guide RNAs (sgRNAs) that could specifically identify 4.1R gene were synthesized and inserted into lentiCRISPRv2 plasmid. RAW264.7 cells were infected with sgRNA?Cas9 lentivirus from 293T cells transfected with the recombinant sgRNA?lentiCRISPRv2 plasmid, and the positive cells were screened using puromycin and the monoclonal cells were obtained. The expression of 4.1R protein in the monoclonal cells was measured by Western blotting, and the mutation site was confirmed by sequence analysis. Result A 4.1R gene knockout RAW264.7 cell line was obtained, which showed a 19?bp deletion mutation in the 4.1R gene sequence and obviously enhanced proliferation. CONCLUSION: We successfully constructed a 4.1R gene knockout macrophage cell line using CRISPR/Cas9 technique, which may facilitate further investigation of the function of 4.1R in macrophages.

Sistemas CRISPR-Cas , Técnicas de Inativação de Genes , Proteínas dos Microfilamentos/genética , RNA Guia/genética , Animais , Camundongos , Células RAW 264.7