Effect of lidamycin on telomerase activity in human hepatoma BEL-7402 cells.
OBJECTIVE
To investigate the effect of lidamycin (LDM) on telomerase activity in human hepatoma BEL-7402 cells under the condition of LDM inducing mitotic cell death and senescence.


METHODS
Chromatin condensation was detected by co-staining with Hoechst 33342 and PI. Cell multinucleation was observed by Giemsa staining and genomic DNA was separated by agarose gel electrophoresis. Fluorescent intensity of Rho123 was determined for mitochondrial membrane potential. MTT assay and SA-beta-gal staining were employed to analyze the senescence-like phenotype. The expression of proteins was analyzed by Western blot. Telomerase activity was assayed by telomerase PCR-ELISA.


RESULTS
Mitotic cell death occurred in LDM-treated cells characterized by unique and atypical chromatin condensation, multinucleation and increased mitochondrial membrane potential. However, no apoptotic bodies or DNA ladders were found. In addition, apoptosis-related proteins remained nearly unaltered. Senescence-like phenotype was identified by increased and elongated size of cells, growth retardation, enhanced SA-beta-gal activity and the changes of senescence-related protein expression. Telomerase activity markedly decreased (P<0.01) in LDM-treated hepatoma BEL-7402 cells.


CONCLUSIONS
Mitotic cell death and senescence could be triggered simultaneously or sequentially after exposure of hepatoma BEL-7402 cells to LDM. The decrease in telomerase activity may play a key role in the defective mitosis and aging morphology. Further investigation of detailed mechanism is needed.
Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100050, China.
Rui-Juan
RJ
Gao
Yue-Xin
YX
Liang
Dian-Dong
DD
Li
Hong-Yin
HY
Zhang
Yong-Su
YS
Zhen
D000617:Aminoglycosides / Q000494:pharmacology*; D000903:Antibiotics, Antineoplastic / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D001399:Azure Stains; D001562:Benzimidazoles; D006528:Carcinoma, Hepatocellular / Q000201:enzymology* / Q000473:pathology*; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D016922:Cellular Senescence / Q000187:drug effects; D002843:Chromatin / Q000378:metabolism; D004273:DNA, Neoplasm / Q000032:analysis; D004305:Dose-Response Relationship, Drug; D053281:Enediynes / Q000494:pharmacology*; D015894:Genome, Human / Q000235:genetics; D006801:Humans; D008113:Liver Neoplasms / Q000201:enzymology* / Q000473:pathology*; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008938:Mitosis / Q000187:drug effects; D010641:Phenotype; D011419:Propidium; D019098:Telomerase / Q000378:metabolism*; D013997:Time Factors; D001616:beta-Galactosidase / Q000378:metabolism
D000617:Aminoglycosides; D000903:Antibiotics, Antineoplastic; D001399:Azure Stains; D001562:Benzimidazoles; D002843:Chromatin; D004273:DNA, Neoplasm; D053281:Enediynes; C058024:C 1027; D011419:Propidium; D019098:Telomerase; D001616:beta-Galactosidase; C017807:bisbenzimide ethoxide trihydrochloride