[Change of Bcl-2 expression and telomerase during apoptosis induced by oridonin on human hepatocelluar carcinoma cells].
OBJECTIVE
To investigate the change of Bcl-2 expression and telomerase during the apoptosis induced by oridonin on human hepatocelluar carcinoma BEL7402 cells.


METHODS
BEL-7402 cells in culture medium were given 8,16,24,32 micromol x L(-1) different concentrations of oridonin. The cell apoptotic rate was detected by flow cytometry (FCM), morphology of cell apoptosis was observed by Hoechst 3325 staining. Bcl-2 and Bax expressions were detected by Western blotting. Reverse transcriptase polymerase chain reaction (RT-PCR) and PCR enzyme-linked immunosorbent assay (ELISA) were used to detect hTERT mRNA expression and telomerase activity.


RESULTS
Oridonin induced BEL-7402 cells apoptosis significantly, and the apoptosis rate was both in time-and dose-dependent manner. Marked morphological changes of cell apoptosis were observed very clearly by Hoechst 33258 staining after the cells exposed to oridonin for 60 hours; Western blotting showed that Bcl-2 expression was down-regulated and Bax expression up-regulated concurrently along with the apoptotic process, and the expression of hTERT mRNA as well as activity of telomerase were decreased concurrently.


CONCLUSIONS
Oridonin could decrease the expression of hTERT mRNA and telomerase activity as well as down-regulation of Bcl-2 and up-regulation of Bax expression during the apoptosis of BEL-7402 cells.
Zhang
Jun-Feng
JF
Department of Liver Transplantation, The Third Affiliated Hospital of Sun Yat-sen University, Guangzhou 510630, China.
Chen
Gui-Hua
GH
Lu
Ming-Qing
MQ
Li
Hua
H
Cai
Chang-Jie
CJ
Yang
Yang
Y
D017209:Apoptosis / Q000187:drug effects*; D006528:Carcinoma, Hepatocellular / Q000378:metabolism* / Q000473:pathology; D045744:Cell Line, Tumor; D004224:Diterpenes / Q000008:administration & dosage / Q000302:isolation & purification / Q000494:pharmacology*; D045786:Diterpenes, Kaurane / Q000008:administration & dosage / Q000302:isolation & purification / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D006801:Humans; D037602:Isodon / Q000737:chemistry; D008113:Liver Neoplasms / Q000378:metabolism* / Q000473:pathology; D010946:Plants, Medicinal / Q000737:chemistry; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000378:metabolism*; D012333:RNA, Messenger / Q000096:biosynthesis / Q000235:genetics; D019098:Telomerase / Q000096:biosynthesis* / Q000235:genetics; D051028:bcl-2-Associated X Protein / Q000378:metabolism
D004224:Diterpenes; D045786:Diterpenes, Kaurane; D019253:Proto-Oncogene Proteins c-bcl-2; D012333:RNA, Messenger; D051028:bcl-2-Associated X Protein; C011959:oridonin; D019098:Telomerase