[Experimental study on apoptosis induced by elemene in glioma cells].
Hongyu Zhou, Jian-kang Shen, Ju-sheng Hou, Yongming Qiu, Qi-zhong Luo
Abstract
Hongyu Zhou, Jian-kang Shen, Ju-sheng Hou, Yongming Qiu, Qi-zhong Luo
Abstract
BACKGROUND & OBJECTIVE: Elemene, isolated from the Chinese medicinal herb Rhizoma Zedoariae, was shown to exhibit antitumor activity. Our previous studies showed that elemene had a markedly antineoplastic activity on glioma. This study was designed to investigate the proliferation inhibitory effect and the apoptosis-inducing activity of elemene on glioma cells. METHODS: The effects of elemene on cell proliferation were studied in vitro by using (3)H-TdR incorporation. The morphological alterations were confirmed by Hoechst 33258/PI staining. The apoptosis was evaluated by flow cytometry analysis and agarose gel electrophoresis. RESULTS: Elemene exhibited a marked antiproliferative effect on rat glioma cell C6 and human glioma cell SHG-44. The fifty percent inhibition concentration (IC(50)) of elemene against glioma cell lines at different time points (D1-D4) by (3)H-TdR incorporation was C6 7.33-11.02 mg/L, SHG-44 13.29-27.16 mg/L. At the same concentration, human glioma cell line SHG-44 was found to be less sensitive to elemene compared to rodent cell line C6. The characteristic nucleolus alternations under fluorescent microscope included condensation of chromatin arranged under the nuclear membrane and apoptotic bodies, with a low nuclear/cytoplasmic ratio. In flow cytometry analysis, a typical subdiploid peak before Phase G(0)/G(1) (apoptotic peak) was detected in DNA frequency distribution histograms. Also the apoptosis in glioma cells was confirmed by DNA ladder formation on gel electrophoresis. CONCLUSION: Elemene exhibited a marked antiproliferative effect on glioma cells, and it could induce apoptosis in vitro.
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BACKGROUND & OBJECTIVE: Elemene, isolated from the Chinese medicinal herb Rhizoma Zedoariae, was shown to exhibit antitumor activity. Our previous studies showed that elemene had a markedly antineoplastic activity on glioma. This study was designed to investigate the proliferation inhibitory effect and the apoptosis-inducing activity of elemene on glioma cells. METHODS: The effects of elemene on cell proliferation were studied in vitro by using (3)H-TdR incorporation. The morphological alterations were confirmed by Hoechst 33258/PI staining. The apoptosis was evaluated by flow cytometry analysis and agarose gel electrophoresis. RESULTS: Elemene exhibited a marked antiproliferative effect on rat glioma cell C6 and human glioma cell SHG-44. The fifty percent inhibition concentration (IC(50)) of elemene against glioma cell lines at different time points (D1-D4) by (3)H-TdR incorporation was C6 7.33-11.02 mg/L, SHG-44 13.29-27.16 mg/L. At the same concentration, human glioma cell line SHG-44 was found to be less sensitive to elemene compared to rodent cell line C6. The characteristic nucleolus alternations under fluorescent microscope included condensation of chromatin arranged under the nuclear membrane and apoptotic bodies, with a low nuclear/cytoplasmic ratio. In flow cytometry analysis, a typical subdiploid peak before Phase G(0)/G(1) (apoptotic peak) was detected in DNA frequency distribution histograms. Also the apoptosis in glioma cells was confirmed by DNA ladder formation on gel electrophoresis. CONCLUSION: Elemene exhibited a marked antiproliferative effect on glioma cells, and it could induce apoptosis in vitro.
Key concepts: Elemene, Apoptosis, Glioma, Flow cytometry, Agarose gel electrophoresis, Molecular biology, Biology, Cell culture