Enhanced Antitumor Effects of Adenoviral-Mediated siRNA against GRP78 Gene on Adenosine-Induced Apoptosis in Human Hepatoma HepG2 Cells
| العنوان: | Enhanced Antitumor Effects of Adenoviral-Mediated siRNA against GRP78 Gene on Adenosine-Induced Apoptosis in Human Hepatoma HepG2 Cells |
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| المؤلفون: | Yan-Qing Ye, Ze-Jin Pu, Jia-Lin Feng, Ling-Fei Wu, Yi-Tian Guo, Meng-Qi Xiang, Guan-You Huang, Wei Deng, Qing-Hua Zhang |
| المصدر: | International Journal of Molecular Sciences, Vol 15, Iss 1, Pp 525-544 (2014) International Journal of Molecular Sciences; Volume 15; Issue 1; Pages: 525-544 International Journal of Molecular Sciences |
| بيانات النشر: | MDPI AG, 2014. |
| سنة النشر: | 2014 |
| مصطلحات موضوعية: | caspase-4, Cell, Apoptosis, Small hairpin RNA, lcsh:Chemistry, RNA interference, RNA, Small Interfering, Endoplasmic Reticulum Chaperone BiP, lcsh:QH301-705.5, Spectroscopy, Heat-Shock Proteins, Membrane Potential, Mitochondrial, Gene knockdown, Liver Neoplasms, General Medicine, Transfection, Hep G2 Cells, Caspases, Initiator, Computer Science Applications, Cell biology, medicine.anatomical_structure, adenosine, Signal transduction, ER stress, medicine.drug, Signal Transduction, Carcinoma, Hepatocellular, Genetic Vectors, glucose-regulated protein 78 (GRP78), Down-Regulation, Antineoplastic Agents, Biology, Catalysis, Article, Adenoviridae, Inorganic Chemistry, medicine, Humans, RNA, Messenger, Physical and Theoretical Chemistry, Molecular Biology, Organic Chemistry, Adenosine, Molecular biology, G1 Phase Cell Cycle Checkpoints, lcsh:Biology (General), lcsh:QD1-999, Apoptosis Regulatory Proteins, Transcription Factor CHOP |
| الوصف: | Our previous studies show that adenosine-induced apoptosis is involved in endoplasmic reticulum stress in HepG2 cells. In this study, we have investigated whether knockdown of GRP78 by short hairpin RNA (shRNA) increases the cytotoxic effects of adenosine in HepG2 cells. The adenovirus vector-delivered shRNA targeting GRP78 (Ad-shGRP78) was constructed and transfected into HepG2 cells. RT-PCR assay was used to determine RNA interference efficiency. Effects of knockdown of GRP78 on adenosine-induced cell viabilities, cell-cycle distribution and apoptosis, as well as relative protein expressions were determined by flow cytometry and/or Western blot analysis. The intracellular Ca2+ concentration was detected by laser scanning confocal microscope. Mitochondrial membrane potential (ΔΨm) was measured by a fluorospectrophotometer. The results revealed that GRP78 mRNA was significantly downregulated by Ad-shGRP78 transfection. Knockdown of GRP78 enhanced HepG2 cell sensitivity to adenosine by modulating G0/G1 arrest and stimulating Bax, Bak, m-calpain, caspase-4 and CHOP protein levels. Knockdown of GRP78 worsened cytosolic Ca2+ overload and ΔΨm loss. Knockdown of caspase-4 by shRNA decreased caspase-3 mRNA expression and cell apoptosis. These findings indicate that GRP 78 plays a protective role in ER stress-induced apoptosis and show that the combination of chemotherapy drug and RNA interference adenoviruses provides a new treatment strategy against malignant tumors. |
| وصف الملف: | application/pdf |
| اللغة: | English |
| تدمد: | 1422-0067 |
| الوصول الحر: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::aae8e6dc968bc0273c9462ecb3531d94Test http://www.mdpi.com/1422-0067/15/1/525Test |
| حقوق: | OPEN |
| رقم الانضمام: | edsair.doi.dedup.....aae8e6dc968bc0273c9462ecb3531d94 |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 14220067 |
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