The PI3K/Akt/mTOR signaling pathway is a key regulator of physiological cell functions which include mobility, difference, apoptosis, proliferation, metabolic process, and autophagy. In CSCs, autophagy plays an essential part in the regulation of drug resistance, self restoration, differentiation, and tumorigenic potential, suggesting autophagy could be a therapeutic target in a subset of cancers. Hence initiating autophagy might abrogate the resistance of CSCs to chemotherapy natural product libraries and can lead to the development of novel therapeutic strategies for the treatment of various cancers. Rottlerin is used as a kinase C delta signaling route inhibitor to verify the natural function of PKC n. It inhibits cell growth and induces apoptosis through mitochondrial membrane depolarization. However, it also acts as an of mitochondrial oxidative phosphorylation in a PKC d independent fashion. Recently, in several human cancer cells, ROT continues to be demonstrated to induce a starvation response, which is a important regulator of autophagy creating its induction. We sought to examine the molecular mechanism by which ROT induces autophagy in pancreatic CSCs, because pancreatic cancer includes pancreatic CSCs. The key aim of the paper is to examine the molecular mechanisms where ROT causes autophagy in pancreatic CSCs. Here we noted that ROT induced early autophagy is principally determined by induction of autophagosomes, transformation of induction of Beclin 1 and Atg7, LC3 I to III and inhibition of Bcl 2 and Bcl XL. Ultimately, ROT induced apoptosis through inhibition of PI3K/Akt/mTOR pathway and activation of caspases. ROT induced apoptosis was increased by Akt1/2 chemical, dominant unfavorable AKT, and rapamycin. Furthermore, inhibition of Beclin 1 and Atg7 increased apoptosisinducing potential of ROT. These studies strongly claim that ROT caused autophagy may play some role as a mechanism against apoptosis. Akt1/2 inhibitors, 3 methyladenine, rottlerin, puromycin, rapamycin, and phenazine methosulfate were from Sigma?Aldrich Corp.. Anti human LC3, Bak, Beclin 1, PKC n, Atg7, Bcl 2, Bcl XL, Bax, cIAP ALK inhibitor 1, Akt, pAkt, mTOR, pmTOR and XIAP were from Cell Signaling Technology. Human pancreatic CSCs were known and described previously. CSCs were grown in DMEM culture medium with 2% B27 Supplement, 1% N2 Supplement, 20 ng/ml individual platelet growth factor, 100 ng/ml epidermal growth factor and 1% antibiotic antimycotic at 37 8C in a atmosphere of five minutes CO2 and 95% air. After drug therapy, total cell lysates were removed using RIPA lysis buffer containing 1-2 protease inhibitor cocktail. Cell lysates were packed and separated on 10. 0-60 Tris?HCl gel. Proteins from the solution were transferred on polyvinylidene difluoride membranes and consequently blocked in blocking buffer and probed with primary antibody at 4 8C for immediately.