PKM2 Knockdown Induces Autophagic Cell Death via AKT/mTOR Pathway in Human Prostate Cancer Cells
| Autor: | Richa Sachan, Prasanta Kumar Dey, Kyungsil Yoon, Nam Deuk Kim, Jae Hyeon Park, Jaewon Lee, Amit Kundu, Hyung Sik Kim, Byung Mu Lee, Mee Young Ahn, In Su Kim |
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| Rok vydání: | 2019 |
| Předmět: |
Male
0301 basic medicine Programmed cell death Physiology Lactate dehydrogenase A Pyruvate Kinase PKM2 lcsh:Physiology lcsh:Biochemistry 03 medical and health sciences Prostate cancer 0302 clinical medicine DU145 Cell Line Tumor Autophagy medicine Humans lcsh:QD415-436 RNA Small Interfering education Mechanistic target of rapamycin Glucose Transporter Type 1 education.field_of_study L-Lactate Dehydrogenase lcsh:QP1-981 biology Chemistry TOR Serine-Threonine Kinases Prostatic Neoplasms Cancer medicine.disease Isoenzymes 030104 developmental biology 030220 oncology & carcinogenesis Cancer cell Cancer research biology.protein RNA Interference Lactate Dehydrogenase 5 Proto-Oncogene Proteins c-akt Signal Transduction |
| Zdroj: | Cellular Physiology and Biochemistry, Vol 52, Iss 6, Pp 1535-1552 (2019) |
| ISSN: | 1421-9778 1015-8987 |
| DOI: | 10.33594/000000107 |
| Popis: | Background/aims Pyruvate kinase M2 (PKM2) is essential for aerobic glycolysis. Although high PKM2 expression is observed in various cancer tissues, its functional role in cancer metabolism is unclear. Here, we investigated the role of PKM2 in regulating autophagy and its associated pathways in prostate cancer cells. Methods Immunohistochemistry was performed to compare the expression level of PKM2 in prostate cancer patients and normal human, whereas expression of PKM2 in several cell lines was also examined by using western blot. PKM2 expression was silenced using various small interfering RNAs (siRNAs). Cell viability was examined using IncuCyte ZOOM™ live cell imaging system. Western blotting and immunofluorescence were performed to investigate the PKM2 knockdown on other cellular signaling molecules. Acridine orange and Monodansylcadaverine staining was performed to check effect of PKM2 knockdown on autophagy induction. High performance thin layer chromatography was carried out to quantify the level of different cellular metabolites (pyruvate and lactate). Colony formation assay was performed to determine the ability of a cells to form large colonies. Results PKM2 was highly expressed in prostate cancer patients as compared to normal human. PKM2 siRNA-transfected prostate cancer cells showed significantly reduced viability. Acridine orange, Monodansylcadaverine staining and western blotting analysis showed that PKM2 downregulation markedly increased autophagic cell death. Results of western blotting analysis showed that PKM2 knockdown affected protein kinase B/mechanistic target of rapamycin 1 pathway, which consequently downregulated the expression of glycolytic enzymes lactate dehydrogenase A and glucose transporter 1. Knockdown of PKM2 also reduced the colony formation ability of human prostate cancer cell DU145. Conclusion To the best of our knowledge, this is the first study to show that PKM2 inhibition alters prostate cancer cell metabolism and induces autophagy, thus providing new perspectives for developing PKM2-targeting anticancer therapies for treating prostate cancer. |
| Databáze: | OpenAIRE |
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