Diabetes was associated with a decrease in podocyte marker proteins

imaging on Day 35 clearly showed that SSHE significantly suppressed the growth of Panc-1 cells without loss of body weight. Additionally, SSHE also suppressed tumor growth of subcutaneously transplanted mouse colon carcinoma LuM1 cells without loss of body weight. -Shogaol causes cell growth retardation and death of pancreatic cancer cells To investigate which constituents of SSHE are active in retarding cell growth and inducing the death of Panc-1 cells, we examined the effect of PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19667322 -shogaol and -gingerol. The results showed that -shogaol reduced the viability of Panc-1 cells with an IC50 value of ~18.8 M. It also caused cell growth retardation and cell death in other pancreatic 11 / 22 Antitumor Activity of Ginger Extract against Pancreatic Cancer Antitumor Activity of Ginger Extract against Pancreatic Cancer Fig 5. ROS production in SSHE-treated Panc-1 cells. Panc-1 cells treated with vehicle alone, 100 g/ml SSHE or 200 g/ml SSHE for 20 h were stained with 10 M H2DCFDA for 10 min and immediately observed under a confocal laser microscope. Panc-1 cells treated as in A were subjected to cytometry. Effect of NAC on SSHE-induced cell death. Panc-1 cells were treated with 200 g/ml SSHE in the presence or absence of 10 mM NAC for 42 h. Cell viability was assessed by a trypan blue dye exclusion test. Bars; SD. As was observed for SSHE, -shogaol decreased the mitochondrial membrane potential, however, it did not activate caspase-3, and zVAD-fmk was ineffective in reversing -shogaol-induced cell death. Nuclear translocation of AIF was not observed, and necrostatin-1 did not show any effect. On the other hand, -shogaol increased the LC3-II/LC3-I ratio, and 3-methyladenine partly recovered the loss of viability caused by -shogaol. -Shogaol increased ROS generation, and the loss of viability caused by -shogaol was reversed by NAC. Treatment of Panc-1 cells with -shogaol resulted in the induction of nuclear shrinkage, cytoplasmic vacuolization, electron-dense mitochondria and empty cytoplasmic spaces. In addition, 0P29 cells were resistant to -shogaol-induced cell PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19666613 death compared with P29mtP29 cells. Together, these results suggest that -shogaol also induces the ROS-mediated autotic death of Panc-1 cells. 13 / 22 Antitumor Activity of Ginger Extract against Pancreatic Cancer Fig 6. Antitumor effect of SSHE in the peritoneal dissemination model of Panc02 cells. Experimental set-up. Bioluminescence imaging on Day 15. Left, IVIS images. Right, Quantitation of photons. FD&C Green No. 3 web Kaplan-Meier survival curve. Body weight. doi:10.1371/journal.pone.0126605.g006 14 / 22 Antitumor Activity of Ginger Extract against Pancreatic Cancer Fig 7. Antitumor effect of SSHE in the orthotopic model of Panc-1 cells. Experimental set-up. Bioluminescence imaging on Day 35. Left, IVIS images. Right, Quantitation of photons. Body weight. Control group; n = 6, SSHE group; n = 5. Presence of potent cell death-inducing component other than shogaol in SSHE To investigate whether -shogaol is the major cell death-inducing component in SSHE, we estimated the concentration of -shogaol in SSHE. To this end, we carried out reversed-phase HPLC followed by the measurement of area corresponding to -shogaol in SSHE. By extrapolating the area to the standard curve, the concentration of -shogaol in SSHE was determined to be approximately 4.6 g in 200 g SSHE. This concentration was not sufficient to achieve full activity of SSHE, which suggested the presence of other active constituent. We then fracti