MedChemExpress Celgosivir onated the constituents in SSHE and examined the cell death-inducing activity of each fraction. The results showed that potent activity was eluted in a few fractions, all of which were ahead of the -gingerol fraction, although the chemical composition of these fractions is presently unknown. -Gingerol fractions showed moderate activity, most likely due to its high content in the extract. -Shogaol fractions showed weak activity. 15 / 22 Antitumor Activity of Ginger Extract against Pancreatic Cancer Discussion PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19666584 The present study demonstrated that the extract of Syussai ginger had potent growthinhibitory and cell death-inducing activity against pancreatic cancer cells including Panc-1 cells. Normal cells such as HUVEC and HPAEpiC were relatively resistant to SSHE PubMed ID:http://www.ncbi.nlm.nih.gov/pubmed/19666601 compared to Panc-1 cells. The extract was also effective under hypoxic conditions, which inevitably develop in all solid tumors to varying degrees and influence the resistance of tumor cells to radiotherapy and conventional chemotherapy. Among various cancers, pancreatic cancer is notorious for its unusually hypoxic microenvironments. Our results showed that SSHE could induce the death of hypoxic Panc-1 cells, which may be a notable characteristic of SSHE for treating pancreatic cancers. In addition to pancreatic cancer, SSHE also induced a marked cell growth retardation and cell death in a variety of tumor cells, which supports the possibility of applying ginger extract and its active constituents for treating cancers. After the treatment of Panc-1 cells with SSHE, the mitochondrial membrane potential was dramatically decreased and the number of annexin V-positive cells increased by 24 h after administration of SSHE. However, little increase in subG1 phase cells, fragmented nuclei and caspase-3 activation, all of which are features of apoptosis, were observed. Moreover, zVAD-fmk failed to rescue SSHE-induced cell death. Because annexin V stains both primary necroptotic and apoptotic cells and mitochondria dysfunction can also be brought about by mitophagy, the autophagy-dependent elimination of mitochondria, we considered the possibility that SSHE-induced cell death was not due to apoptosis. Additionally, necroptosis was excluded because necrostatin-1 could not ameliorate the cell death induced by SSHE. We then explored the possibility of autophagic cell death and noticed many cytoplasmic vacuoles and LC3 puncta in the SSHE-treated Panc-1 cells at early stages. SSHE treatment dramatically increased the ratio of LC3-II/LC3-I and caused loss of SQSTM1/p62. SSHE activated AMPK and inhibited mTOR. 3-Methyladenine and chloroquine partially rescued SSHE-induced cell death. All of these data suggested that autophagy was occurring in the SSHE-treated cells. However, a close look at the SSHE-treated cells revealed several features that are not observed in classical type autophagy, including nuclear shrinkage, focal membrane rupture, electrondense mitochondria, empty vacuoles and focal perinuclear swelling. Eventually, it appeared that these morphological 
features coincided well with the recently discovered form of cell death, “autosis”. Thus, we presently consider that the SSHE-induced cell death of Panc-1 cells is mainly due to caspase-independent, autotic cell death rather than due to apoptosis and necroptosis. Several lines of evidence indicate that ROS, more specifically mitochondria-derived ROS, are the inducer of autophagy upon various stresses such as nutrient deprivation. Our data