MG-132 inhibits telomerase activity, induces apoptosis and G(1) arrest associated with upregulated p27kip1 expression and downregulated survivin expression in gastric carcinoma cells.

Abstract

Ubiquitin-proteasome pathway (UPP) is the major system for the selective degradation of cellular proteins that play key roles in cellular processes. Previous study indicated that ubiquitin-proteasome inhibitor MG-132 could inhibit growth of some carcinoma. However, anti-carcinoma mechanism of MG-132 is unclear. Our objective was to investigate mechanisms of growth inhibitory effect of MG-132 on gastric carcinoma cells. Gastric carcinoma cell SGC-7901 was treated with ubiquitin-proteasome inhibitor MG-132. Cell growth suppression was evaluated with 3-(4,5-dimethylthiazole-2-yl)-2,5-diphenyl tetrazolium bromide (MTT) assay. DNA synthesis was evaluated by (3)H-thymidine ((3)H-TdR) incorporation. Activity of telomerase was examined by telomeric repeat amplification protocol (TRAP) PCR-ELISA. Cell cycle and apoptosis were detected by flow cytometry (FCM). DNA fragment analysis was used to confirm the presence of apoptosis. Expression of p27kip1 and survivin was detected using the western blot method. After exposed to MG-132, the growth and value of (3)H-TdR incorporation of gastric carcinoma cells were obviously inhibited. TRAP PCR-ELISA showed that light absorption of cells gradually decreased after exposed to 5 microM of MG-132 for 24, 48, 72 and 96 h (P < 0.01). The percentage of cells at G(0)/G(1) phase was increased and that at S and G(2)/M phase was decreased (P < 0.01). The ratio of apoptotic cells treated with 5 microM MG-132 for 96 h was 53.7 +/- 6.4%. Agarose electrophoresis showed marked ladders. Moreover, expression of p27kip1 of cells was increased and expression of survivin was decreased. Our results suggest that MG-132 inhibits telomerase activity, induces apoptosis and G(1) arrest which is associated with upregulated p27kip1 expression and downregulated survivin expression in gastric carcinoma cells.

• Authors: Weiguo Zhang, Qiang Tong, Shengbao Li, Xiaohu Wang, Qiang Wang
• Journal: Cancer investigation (Cancer Invest) Vol. 26 Issue 10 Pg. 1032-6 (Dec 2008) ISSN: 1532-4192 [Electronic] England
• PMID: 19093261 (Publication Type: Journal Article)

Chemical References

• Antineoplastic Agents
• BIRC5 protein, human
• CDKN1B protein, human
• Inhibitor of Apoptosis Proteins
• Intracellular Signaling Peptides and Proteins
• Leupeptins
• Microtubule-Associated Proteins
• Survivin
• Cyclin-Dependent Kinase Inhibitor p27
• Telomerase
• benzyloxycarbonylleucyl-leucyl-leucine aldehyde

Topics

• Antineoplastic Agents (therapeutic use)
• Apoptosis (drug effects)
• Cell Division (drug effects)
• Cell Line, Tumor
• Cyclin-Dependent Kinase Inhibitor p27
• DNA Replication (drug effects)
• Down-Regulation
• Flow Cytometry
• G1 Phase (drug effects, physiology)
• Gene Expression Regulation, Neoplastic
• Humans
• Inhibitor of Apoptosis Proteins
• Intracellular Signaling Peptides and Proteins (genetics)
• Leupeptins (pharmacology)
• Microtubule-Associated Proteins (genetics)
• Stomach Neoplasms (drug therapy, genetics, pathology)
• Survivin
• Telomerase (antagonists & inhibitors)
• Tumor Cells, Cultured