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3 Fundamental Details About PR-171 Outlined

Treatment find more groups received 250 ��L lentivirus by intravenous injection into the tail vein every 24 hr for 3 weeks. Upon termination, each mouse was weighted and tumors were harvested for Western blot analysis and quantitative real-time RT-PCR. Data were expressed as mean �� SD. The difference between two groups was analyzed by the Student's t-test using SPSS software (Release 11.0, SPSS Inc.). A value of P < 0.05 was considered as statistical significance. In order to determine whether NUPR1 was involved in the regulation of tumorigenesis of human NSCLC, we assessed the NUPR1 expression level in NSCLC tissue specimens and corresponding peritumoral lung tissues. As shown in (Fig. 1A), NSCLC samples showed up-regulation of NUPR1 as compared with peritumoral lung tissues (P <0.01). The expression levels of NUPR1 were examined in human NSCLC cells A549, SK-MES-1, 95-D, NCI-H460, NCI-H1650, H1299, and human fetal lung fibroblast cell line MRC-5 by real-time quantitative RT-PCR analysis. The result indicated that NUPR1 expression was upregulated in the entire human nonsmall cell A lung cancer cell lines examined (Fig. 1B). The expression levels of NUPR1 mRNAs in A549, SK-MES-1, 95-D, NCI-H460 and NCI-H1650 were relatively lower than those in H1299 cells. Therefore, we selected the H1299 cell line for use in the following in vitro assays. To determine the impact of NUPR1 expression on the growth and proliferation of human lung cancer cells, we generated lentiviruses that expressed the NUPR1 RNAi and control RNAi, respectively. The success of gene silencing is dependent on the choice of appropriate target sequences and a sensitive assay to select the RNAi that suppresses <a href="http://www.selleckchem.com/products/azd9291.html">AZD9291 datasheet gene YES1 expression most efficiently. The effect of NUPR1 RNAi down-regulation of NUPR1 expression was tested in human lung cancer cell line H1299 (Fig. 2A). After the cells were infected with NUPR1 RNAi or control RNAi for 4 days, the mRNA level of cells infected with NUPR1 RNAi was down-regulated by 98.4% compared to cells infected with control RNAi, and there is significant difference between the NUPR1 RNAi and the control RNAi infected cells (P<0.01, Fig. 2B). Thus, NUPR1 RNAi could down-regulate NUPR1 expression level efficiently. To investigate the possible function of NUPR1 in the proliferation of human lung cancer H1299 cells, the dynamics of the control RNAi and the NUPR1-specific RNAi cell proliferation were determined by cell count assay. Following a 5-day period, the proliferation of control RNAi NUPR1 cells showed strong proliferation. However, the proliferation of NUPR1 RNAi cells was much slower, as demonstrated by that significantly lower numbers of NUPR1 RNAi H1299 cells were observed on Day 2, and the number of cells were reduced by about 90.5% as compared with that in control groups on Day 5 (P < 0.01) (Fig. 3A). The effect of NUPR1 down-regulation on the potential cells growth was examined by colony formation assay in H1299 cells. As shown in Fig.</div>
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