PKC-δ在TRAIL与阿霉素联合诱导骨肉瘤细胞株MG-63凋亡中的作用

PKC-δ在TRAIL与阿霉素联合诱导骨肉瘤细胞株MG-63

凋亡中的作用

李广章;刘长剑;汤欣

【期刊名称】《中国骨与关节杂志》 【年(卷),期】2011(010)003

【摘要】Objective To preliminarily observe the synergistic effect of TNF-related apoptosis-inducing ligand (TRAIL) and low doses of adriamycin (ADM) on the apoptosis of cell line MG-63 in osteosarcoma and probe the function of protein kinase C-δ (PKC-δ) in this process.Meanwhile to discuss the function of PKC-δ through the effect produced by phorbol-12-myristate-13-acetate (PMA) which is the activator of PKC and Rottlerin which is the specific inhibitor of PKC-δ on the apoptosis of cell line MG-63 in osteosarcoma.Methods The morphologic changes of the apoptosis of cell line MG-63 conducted with TRAIL and ADM conjointly were observed through inverted phase contrast microscope, and meanwhile the changes of messenger ribose nucleic acid (mRNA) expression of PKC-δ were tested by reverse transcription-polymerase chain reaction (RT-PCR).Flow cytometry (FCM) was used to test the changes of apoptosis rate in the cell group conducted conjointly with PMA and Rottlerin.Results Some MG-63 cells got round and began to float after 24 hours' treating with 5μg/ml ADM and 500ngml TRAIL.Cellular fragments of varying sizes in the cytoplasm can be

seen.The mRNA expression of PKC-δ in the group conjointly applied with TRAIL and ADM increased obviously.Compared with the group conducted with only 1 substance and control group, significant difference existed (P＜0.01).After 12 hours.Rottlerin with combined medications could decrease the apoptosis rate while PMA with combined medications could increase it (P＜0.01).Conclusions TRAIL and low dose ofADM conducted conjointly can promote the apoptosis of cell line MG-63.Rottlerin may inhibit the apoptosis of cell line MG-63 inducted jointly by TRAIL and ADM.PMA plays a positive role.PKC-δ may participate in the process of apoptosis and play a promoting role.%目的 初步观察肿瘤坏死因子相关凋亡诱导配体(TRAIL)与低剂量阿霉素(ADM)联合应用对人骨肉瘤细胞株MG-63细胞凋亡的影响,探讨蛋白激酶C-δ (PKC-δ)在其中的作用,并用PKC激活剂佛波脂(PMA)和PKC-δ特异性抑制剂Rottlerin对这一凋亡过程的影响来探讨PKC-δ在其中的作用.方法 倒置相差显微镜观察TRAIL与ADM联合应用后MG-63细胞凋亡形态的变化,同时逆转录-聚合酶链反应法检测PKC-δ的mRNA表达变化.流式细胞仪(FCM)检测PMA和Rottlerin作用联合组细胞后凋亡率的变化.结果 5μg/ml的ADM与500ng/ml的TRAIL联合作用24h后,部分MG-63细胞变圆、悬浮,可见大小不等的细胞碎片.TRAIL和ADM联合应用组PKC-δ的mRNA的表达明显增加,与单用组和对照组有显著性差异(P＜0.01).Rottlerin与联合用药作用12h后细胞的凋亡率降低,PMA则能使凋亡率增加(P＜0.01).结论 TRAIL与低剂量ADM联合作用可以促进MG-63细胞凋亡.Rottlerin能抑制TRAIL和ADM联合诱导MG-63细

胞凋亡,PMA则起促进作用,PKC-δ可能参与了该凋亡过程并起促进作用. 【总页数】5页(281-284,310)

【关键词】肿瘤坏死因子相关凋亡诱导配体;阿霉素;蛋白激酶C-δ;骨肉瘤;凋亡 【作者】李广章;刘长剑;汤欣

【作者单位】116011,大连医科大学附属第一医院;116011,大连医科大学附属第一医院;116011,大连医科大学附属第一医院 【正文语种】中文 【中图分类】 【文献来源】

https://www.zhangqiaokeyan.com/academic-journal-cn_chinese-journal-bone-joint_thesis/0201233938204.html 【相关文献】

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