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Several Catabolic Proteins are Down-regulated in Starvation Induced Autophagy in HeLa Cells
Dan-Hui Liu, Shan-Shan Han, Lu-Yun Li, Ye-Min Qu, Hong Lin, Jie Wen, Jian-Xin Lü*
Institute of Molecular & Cellular Medicine, Zhejiang Provincial Key Laboratory of Medical Genetics, Institute of Proteomics, Wenzhou Medical College, Wenzhou 325035, China
Abstract: Autophagy is a conversed catabolic pathway that plays an important role in maintaining cell homeostasis by degrading damaged cytosolic components and redundant proteins in eukaryotic cells. Autophagy plays a protective role against various human diseases， including cancer. Although many autophagy related proteins have been investigated， there are many unsolved problems about the mechanism of the occurrence and regulation of autophagy. In this study， autophagy was induced in HeLa cells by starvation. The occurrence of autophage was confirmed with transmission electron microscopy and LC3-I conversion， which are hallmarks of autophagy. We used 2-DE coupled with LC-MS/MS to analyze the differential protein expression in starved HeLa cells. We found fructose-bisphosphate aldolase A， GAPDH and ATP synthase O subunit were significantly decreased in starvationinduced autophagy. This was confirmed with real-time quantitative PCR. To address if these proteins were directly related with autophagy， HeLa cells were pretreated with the autophagy inhibitor 3-MA， and the total RNA were extracted for real-time quantitative PCR after starvation. The data revealed 3-MA can rescue the mRNA decrease of fructose-bisphosphate aldolase A， GAPDH and ATP synthase O subunit. Taken together， our results indicate that these proteins may be involved in the regulation of autophagy， and the mechanism need further investigation.
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