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  • A MTT analysis of AsPC pLXSN Adh

    2020-03-24

    A) MTT analysis of AsPC-1 + pLXSN (Adh.) HG-9-91-01 plated with the different NAX compounds. In Panel B, the IC50s are presented graphically. All of the experiments indicated in this figure were performed on the same day. These experiments were repeated 3 times and similar results were obtained.
    have significant activity on any of the three PDAC lines. The means and standard deviations are presented in Figs. 2–4. The IC50 values and the fold-difference of the IC50s in comparison to BBR are presented in Tables 1–3, as well as, statistical analysis of the differences of the IC50s.
    3.3. Effects of BBR and NAX compounds on colony formation in MIA-PaCa-2 cells
    As an additional approach to examine the effects of the NAX compounds cell growth, we determined their effects on colony formation in the presence and absence of chemotherapeutic drugs and they are presented in Fig. 5. The more effective NAX com-pounds, which we determine to be more effective by MTT analysis were also more potent in suppressing colony formation (Fig. 5). Essentially, no growth was observed after 200 nM NAX053 or NAX060 treatment, while this same concentration of BBR only reduced colony formation about 50% in BBR treated cells. All the cells and NAX053, NAX060 and BBR were set up the same day and incubated for the same period of time. In contrast, incubation of the cells with NAX054 did not have any effects on the colony formation ability of the MIA-PaCa-2 + pLXSN (Adh.) cells.
    3.4. Effects of BBR and NAX compounds on MIA-PaCa-2 cells containing introduced WT-TP53
    Fig. 4. Effects of the Nutraceuticals Berberine And NAX Compounds on the Proliferation and IC50s of PANC-28 + pLXSN (Adh.) Cells. Panel A) MTT analysis of PANC-28 + pLXSN (Adh.) cells plated with the different NAX compounds. In Panel B, the IC50s are presented graphically. All of the experiments indicated in this figure were performed on the same day. These experiments were repeated 4 times and similar results were obtained.
    Table 1 Effects of Berberine and NAX Compounds on MIA-PaCa-2 + pLXSN (Adh.) Cells.a
    Compound IC50
    Fold Difference of IC50 with BBR Statistical Significance
    a Determined by MTT Analysis as presented in Abrams et al. (2018).
    Table 2 Effects of Berberine and NAX Compounds on AsPC-1 + pLXSN (Adh.) Cells.a
    Compound IC50
    Fold Difference of IC50 with Berberine Statistical Significance
    a Determined by MTT Analysis as presented in Abrams et al. (2018).
    Table 3 Effects of Berberine and NAX Compounds on PANC-28 + pLXSN (Adh.) Cells.a
    Compound IC50
    Fold Difference of IC50 with Berberine Statistical Significance
    a Determined by MTT Analysis as presented in Abrams et al. (2018).
    Fig. 5. Effects of Nutraceutical BBR and NAX Compounds on Colony Formation in MIA-PaCa-2 + pLXSN (Adh.) Cells. The colony formation was determined and normalized to untreated samples. These experiments were repeated 3 times.
    4. Discussion
    BBR is consumed by millions of people for various diseases. Various attempts have been tried to improve the effectiveness of BBR in suppressing various biological/clinical problems. In our studies, we examined the effects of a panel of modified BBRs (NAX compounds) on three different PDAC cell lines. Some of the NAX compounds inhibited proliferation more than other NAX compounds and the parental BBR both in terms of MTT assays and IC50 determination and also in colony formation assays. Interestingly, the effects of the NAX compounds were similar on all three PDAC cell lines.
    MIA-PaCa-2 cells have a gain of function mutation at TP53 (Deer et al., 2010). We have examined previously the effects of BBR on these cells (Abrams et al., 2018). The MIA-PaCa-2 + WT-TP53 (Adh.) were approximately 3-fold more sensitive to BBR than MIA-PaCa-2+ pLXSN (Adh) cells which lacked introduced WT-TP53 (Abrams et al., 2018). In this current manuscript the effects of the modified NAX compounds were examined the same day as the natural BBR on MIA-PaCa-2 + pLXSN cells. An overall similar patter of sensitivity to the modified NAX compounds was observed in the MIA-PaCa-2 + WT-TP53 (Adh.) as in the MIA-PaCa-2 + pLXSN (Adh.) cells. Thus, cells containing WT-TP53 were sensitive to the NAX compounds.