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  • br F br hBMSCs miR NC br

    2020-08-18


    F
    hBMSCs-miR-NC
    expression 2.0
    VEGF
    protein 1.0
    GAPDH
    Relative
    -miR -miR
    NC
    mimic
    downregulating ADAM9 (Figure 11). The observations and data highlight the potential of miR-126-3p as a target for pancreatic cancer treatment. However, this study has its own shortcomings especially with small sample size. Further study concentrating on selection of two or more cell lines and selection of more animal experiments might increase the credibility and implication of the results.
    MATERIALS AND METHODS
    Ethics Statement
    This study was approved by the Institutional Review Board and the Institutional Animal Care and Use Committee of School of Life Sci- 
    Microarray Analysis
    Through the retrieval of the GEO data-
    base (https://www.ncbi.nlm.nih.gov/geo/), four
    pancreatic cancer correlated gene chips
    abnormal expressed genes in pancreatic cancer
    ki67 were screened from the four gene expression
    chips. Affy installation package in R language46
    was employed in order to standardize the pre-
    treatment of the chip Resolvin E1 data. The limma package47 was used for differential gene screening with jlog2 FCj > 1.0, and adj.P.Val (corrected p value) < 0.05 was set as the threshold. Next, the expres-sion thermal map of differential genes was constructed. The Calculate and draw custom Venn diagrams (http://bioinformatics.psb.ugent. be/webtools/Venn/) were used to compare the differential genes in four gene chips. The GEPIA database (http://gepia. cancer-pku.cn/)48 was employed to verify the expression of differen-tial genes and analyze the correlation between gene expression and survival conditions. TargetScan (http://www.targetscan.org/vert_71/), miRSearch (http://www.exiqon.com/microrna-target-prediction),
    Molecular Therapy: Nucleic Acids
    A
    Tumorgrowth 500
    hBMSCs-miR-126-3p mimic hBMSCs-miR-
    hBMSCs-miR-
    E
    C
    Tumor weight (g)
    D
    Relative  2
    GAPDH
    - -miR -miR -3p NC -3p mimic hBMSCs
    hBMSCs
    protein 0.5
    Relative 0.0
    Figure 10. Overexpressed miR-126-3p Inhibited Tumor Growth and Metastasis of Pancreatic Cancer
    (A) The effect of BMSCs-miR-126-3p mimic on the growth of the tumor detected by xenograft in nude mice. (B) Effect of BMSCs-miR-126-3p mimic on tumor volume detected by xenograft in nude mice. (C) Effect of BMSCs-miR-126-3p mimic on the quality of the tumor detected by xenograft in nude mice. (D) Expression of miR-126-3p and ADAM9 after BMSCs-miR-126-3p mimic treatment determined by qRT-PCR. (E and F) Protein expression of ADAM9, COX-2, and MMP-14 determined by western blot analysis (photograph of protein expression of ADAM9, COX-2, and MMP-14, E, and relative protein expression of ADAM9, COX-2, and MMP-14, F). *p < 0.05, compared with the BMSCs-miR-126-3p NC group. The data in the map were all measurement data and expressed by mean ± SD. (A) Data were analyzed by repeated-measurement variance of analysis. (C–E) Data were analyzed by independent sample t test for statistical analysis. n = 12. BMSC, human bone mesenchymal stem cell; COX-2, cyclo-oxygenase-2; miR-126-3p, microRNA-126-3p; MMP-14, matrix metalloproteinase-14; NC, negative control.
    miRTarBase (http://mirtarbase.mbc.nctu.edu.tw/php/search.php), miRWalk (http://mirwalk.umm.uni-heidelberg.de/), and mirDIP (http://ophid.utoronto.ca/mirDIP/), five miRNA-mRNA relation prediction databases, were applied to predict the target miRNA of differentially expressed genes and compare predicted results of five miRNAs. The miRNA expression chip GEO: GSE28955 of pancreatic cancer was analyzed by R language using the same method of gene expression chip. Differentially expressed miRNAs in pancreatic can-cer tissues were screened and compared with the target miRNAs of the differential genes.
    Cell Culture and Tissue Collection
    Six pancreatic cancer cell lines, SW1990, Capan-1, AsPC-1, MIAPaCa-2, PANC-1, and PC-3, and human normal pancreatic cell line HPC-Y5 were purchased from Shanghai YanHui Biotech-nical (Shanghai, China).
    Specimens from 28 patients with pancreatic cancer and 32 patients with pancreatitis who had undergone surgical resection procedures at the School of Life Science, College of Health Sciences, Jiangsu Normal University from January 2018 to May 2018 were recruited for the purposes of the study. Five pancreatic cancer tissues and five pancreatitis tissues were promptly excised using sharp knives within a period of 5 min after the specimens had been removed from the patients. All patients enrolled into the study were yet to be administered chemotherapy, radiotherapy, or immunotherapy prior