摘要
目的观察透骨草提取物对人卵巢癌SKOV-3细胞凋亡和p53正向细胞凋亡调控因子(PUMA)表达的影响,探讨PUMA蛋白和mRNA表达在透骨草提取物诱导SKOV-3细胞凋亡中的作用。方法采用透射电镜观察透骨草提取物对SKOV-3细胞超微结构的影响,采用免疫组织化学方法检测透骨草提取物对SKOV-3细胞PUMA蛋白表达水平的影响:采用反转录聚合酶链反应(RT-PCR)检测透骨草提取物对SKOV-3细胞PUMA mRNA表达水平的影响。结果透射电镜观察结果显示,37.8μg/mL透骨草提取物作用24h的SKOV-3细胞胞膜皱缩,表面突起和微绒毛减少,染色质凝聚、边缘化,细胞核裂解为碎块,产生凋亡小体;免疫组织化学染色结果显示,37.8μg/mL透骨草提取物作用的SKOV-3细胞PUMA蛋白表达明显高于对照组(P<0.05);RT-PCR方法检测结果显示,37.8μg/mL透骨草提取物作用的SKOV-3细胞PUMA mRNA表达明显高于对照组P<0.05)。结论透骨草提取物可以诱导SKOV-3细胞凋亡,PUMA在透骨草提取物诱导SKOV-3细胞凋亡的过程中具有一定作用,诱导PUMA表达可能是透骨草提取物抗卵巢癌作用的机制之一。
Objective To detect the effects of Phryma leptostachyaL.var.asiatica Hara extract on apoptosis and the expression of PUMA of human ovarian cancer SKOV-3cells and to reveal the effect of PUMA protein and PUMA mRNA on inducing the apoptosis of SKOV-3cells.Methods The transmission electronic microscope was used to observe the effect of Phryma leptostachya L.var.asiatica Hara extract on the ultrastructure of SKOV-3cells.The immunohistochemistry technique was used to detect the effect of Phryma LeptostachyaL.var.asiatica Hara Extract on the expressions of PUMA protein of SKOV-3cells.RT-PCR was used to detect the effect of Phryma LeptostachyaL.var.asiatica Hara extract on the expressions of PUMA mRNA of SKOV-3cells.Results The results of transmission electronic microscope showed that SKOV-3cells shrank,the surface processes and microvillus decreased,chromatin condensed and marginalized,nucleus fragmented and apoptotic bodies appeared after SKOV-3cells were treated with 37.8μg/mL Phryma leptostachyaL.var.asiatica Hara extract.The results of immunohistochemistry technique showed that the expression of PUMA protein in SKOV-3cells treated with37.8μg/mL Phryma leptostachyaL.var.asiatica Hara extract increased significantly compared with the control group（P0.05）.RT-PCR showed that the expression of PUMA mRNA in SKOV-3cells treated with 37.8μg/mL Phryma leptostachyaL.var.asiatica Hara extract increased significantly compared with the control group（P0.05）.Conclusion Phryma leptostachyaL.var.asiatica Hara extract can induce the apoptosis of SKOV-3cells.PUMA play important roles in inducing the apoptosis of SKOV-3cells.Inducing the expression of PUMA may be one of the mechanism for anti-ovarian cancer effect of Phryma leptostachya L.var.asiatica Hara extract.
出处
《山西医药杂志》
CAS
2014年第20期2371-2373,共3页
Shanxi Medical Journal
基金
大连市科学技术基金计划(2010E13SF144)
