title: Rottlerin induces autophagy and apoptosis in prostate cancer stem cells via PI3K/Akt/mTOR signaling pathway.
abstract: Autophagy plays an important role in cellular homeostasis through the disposal and recycling of cellular components. Cancer stem cells (CSCs) play major roles in cancer initiation, progression, and drug resistance. Rottlerin (Rott) is an active molecule isolated from Mallotus philippinensis, a medicinal plant used in Ayurvedic Medicine for anti-allergic and anti-helminthic treatments, demonstrates anticancer activities. However, the molecular mechanisms by which it induces autophagy in prostate CSCs have not been examined. The main objective of the paper was to examine the molecular mechanisms by which Rott induces autophagy in prostate CSCs. Autophagy was measured by the lipid modification of light chain-3 (LC3) and the formation of autophagosomes. Apoptosis was measured by flow cytometer analysis. The Western blot analysis was used to examine the effects of Rott on the expression of PI3K, phosphorylation of Akt, phosphorylation of mTOR, and phosphorylation of AMPK in pros CSCs. RNAi technology was used to inhibit the expression of Beclin-1 and ATG-7. Rott induced the lipid modification of light chain-3 (LC3) and the formation of autophagosomes after 24h of Rott treatment in prostate CSCs. Rott-treated prostate CSCs induced transition from LC3-I to LC3-II, a hall mark of autophagy. Rott also induced the expression of Atg5, Atg7, Atg12 and Beclin-1 proteins during autophagy. The knock-down of Atg7 and Beclin-1 blocked Rott-induced autophagy. Furthermore, Rott induced AMPK phosphorylation was blocked by 3-MA, Baf and CHX. In addition, inhibition of AMPK expression by shRNA blocked Rott induced autophagy. In conclusion, a better understanding of the biology of autophagy and the pharmacology of autophagy modulators has the potential for facilitating the development of autophagy-based therapeutic interventions for prostate cancer.
authors.affiliation: Department of Pharmacology, Toxicology and Therapeutics, and Medicine, The University of Kansas Medical Center, 3901 Rainbow Boulevard, Kansas City, 66160, USA. Electronic address: dkumar2@kumc.edu.
authors.forename: Dhruv
authors.initials: D
authors.lastname: Kumar
authors.affiliation: Department of Pathology and Laboratory Medicine, The University of Kansas Medical Center, 3901 Rainbow Boulevard, Kansas City, KS 66160, USA. Electronic address: sshankar@kumc.edu.
authors.forename: Sharmila
authors.initials: S
authors.lastname: Shankar
authors.affiliation: Department of Pharmacology, Toxicology and Therapeutics, and Medicine, The University of Kansas Medical Center, 3901 Rainbow Boulevard, Kansas City, 66160, USA. Electronic address: rsrivastava@kumc.edu.
authors.forename: Rakesh K
authors.initials: RK
authors.lastname: Srivastava
mesh_terms: D000098:Acetophenones / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000494:pharmacology; D017209:Apoptosis / Q000187:drug effects; D001343:Autophagy / Q000187:drug effects; D001578:Benzopyrans / Q000494:pharmacology*; D015153:Blotting, Western; D045744:Cell Line, Tumor; D005434:Flow Cytometry; D006801:Humans; D008297:Male; D058539:Phosphatidylinositol 3-Kinase / Q000378:metabolism*; D011471:Prostatic Neoplasms / Q000188:drug therapy; D015398:Signal Transduction / Q000187:drug effects*; D058570:TOR Serine-Threonine Kinases / Q000378:metabolism*
chemical_list: D000098:Acetophenones; D000970:Antineoplastic Agents; D001578:Benzopyrans; C085746:rottlerin; C546842:MTOR protein, human; D058539:Phosphatidylinositol 3-Kinase; D058570:TOR Serine-Threonine Kinases
keywords: 4′,6-diamidino-2-phenylindole; 5′ adenosine monophosphate-activated protein kinase; AMPK; Akt; Atg; Autophagy; Baf; CHX; CSCs; DAPI; FBS; LC3; PI3K; Rott; autophagy related gene; bafilomycin; cancer stem cells; cycloheximide; fetal bovine serum; mTOR; mammalian target of rapamycin; microtubule-associated protein 1 light chain 3; phosphatidylinositide 3-kinases; rottlerin