Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Marigold Supercritical Extract as Potential Co-adjuvant in Pancreatic Cancer: The Energetic Catastrophe Induced via BMP8B Ends Up With Autophagy-Induced Cell Death

doi: 10.3389/fbioe.2019.00455

Figure Lengend Snippet: BMP8B is a molecular target of marigold SFE. (A) Microarray data of differentially expressed genes after treatment of MiaPaCa-2 pancreatic cancer cell line with 30 and 70 μg/ml marigold SFE for 48 h. Data represent the value of the most significant probe for three independent experiments for each condition. Genes with a statistically significant difference ( P value < 0.05) and more than two-fold absolute change variation compared to control (DMSO) are shown. (B) Validation of the upregulation of BMP8B after marigold SFE treatment in two pancreatic cancer cell lines, MiaPaca-2 and Panc-1. Non-treated cells were kept as controls. Data represent the mean ± S.E.M. of three independent experiments, each one performed in triplicate. Asterisks indicate statistical differences in treated cells in comparison with control (DMSO). ** P < 0.05, **** P < 0.001 indicate statistically significant differences refered to the control C.

Article Snippet: Quantitative polymerase chain reaction (qPCR) was performed in the 7900HT Real-Time PCR System (Life Technologies) using the VeriQuest SYBR Green qPCR Master Mix (Affymetrix, Santa Clara, CA, USA), and Taqman probes were used: Hs01629120_s1, Hs01029413_m1, Hs00245183_m1, and Hs99999901_s1 for BMP8B, TFAP2A, ZFP36L1 , and 18S, respectively; or oligos in the case of the epithelial-to-mesenchymal transition (EMT), stemness and endoplasmic reticulum (ER) stress markers ( displays the list and sequences of the primers used).

Techniques: Microarray

Journal: Frontiers in Bioengineering and Biotechnology

Article Title: Marigold Supercritical Extract as Potential Co-adjuvant in Pancreatic Cancer: The Energetic Catastrophe Induced via BMP8B Ends Up With Autophagy-Induced Cell Death

doi: 10.3389/fbioe.2019.00455

Figure Lengend Snippet: Depletion of BMP8B counteracts the effect of marigold SFE on the inhibition of mitochondrial oxidative phosphorylation. (A) Mitochondrial respiration analysis by flux analysis of the OCR in MiaPaca-2 transfected with si-RNA pools against BMP8B or si-RNA-scramble, in non-treated cells and marigold SFE-treated cells (1 × IC 50 , 2 × IC 50 ). Lower panels: basal respiration rate, spare respiratory capacity, ATP production, and proton leak. (B) Quantitative real-time PCR (qRT-PCR) validation of the partial inhibition of BMP8B by means of si-RNA pools against BMP8B in MiaPaca-2 cells. (C) Effect of marigold SFE on the expression levels of BIP and CHOP in MiaPaca-2 cells transfected with si-RNA pools against BMP8B or si-RNA-scramble. (D) Mitochondrial respiration analysis by flux analysis of the OCR in Panc-1 cells transfected with si-RNA pools against BMP8B or si-RNA-scramble, in non-treated cells and marigold SFE-treated cells (1 × IC 50 , 2 × IC 50 ). (E) qRT-PCR validation of partial inhibition of BMP8B by means of si-RNA pools against BMP8B in Panc-1 cells. (F) Effect of marigold SFE on the expression levels of BIP and CHOP in Panc-1 cells transfected with si-RNA pools against BMP8B or si-RNA-scramble. *Indicates statistical differences referred to control non-treated cell transfected with siRNA scramble: * P < 0.01; ** P < 0.05, *** P < 0.005, **** P < 0.001. # Indicates statistical differences between siRNA BMP8B and siRNA scramble transfected cells for each condition: # P < 0.05, ## P < 0.01, ### P < 0.005, #### P < 0.001.

Article Snippet: Quantitative polymerase chain reaction (qPCR) was performed in the 7900HT Real-Time PCR System (Life Technologies) using the VeriQuest SYBR Green qPCR Master Mix (Affymetrix, Santa Clara, CA, USA), and Taqman probes were used: Hs01629120_s1, Hs01029413_m1, Hs00245183_m1, and Hs99999901_s1 for BMP8B, TFAP2A, ZFP36L1 , and 18S, respectively; or oligos in the case of the epithelial-to-mesenchymal transition (EMT), stemness and endoplasmic reticulum (ER) stress markers ( displays the list and sequences of the primers used).

Techniques: Inhibition, Transfection, Real-time Polymerase Chain Reaction, Quantitative RT-PCR, Expressing