摘要
目的 观察二氟甲基鸟氨酸 (DFMO)及反义bcl 2调控bcl 2基因表达对HL6 0细胞凋亡的诱导作用。方法 构建反义bcl 2逆转录病毒重组体 ,建立产病毒细胞系 ,转染HL6 0细胞 ,用形态观察、生长曲线、FCM分析、集落形成、DNA电泳图谱、分子杂交及免疫组化等方法研究细胞生长特性及细胞凋亡诱导。结果 成功地构建了反义bcl 2逆转录病毒重组体及产病毒细胞系。以此转染HL6 0细胞 ,引起了bcl 2mRNA及蛋白表达下降 ,但生长速率、细胞周期分布及鸟氨酸脱羧酶 (ODC)基因表达水平与亲本细胞比较差别不大 ;无明显的细胞凋亡 ,仅集落形成能力有所减弱。用小剂量DFMO处理反义bcl 2转染的细胞 ,不仅生长受到明显抑制 ,而且可诱导细胞凋亡。结论 DFMO与反义bcl 2对HL6 0细胞增殖抑制及凋亡诱导有协同作用。抑制bcl 2表达能提高肿瘤细胞对DFMO作用的敏感性。
Objective To study the combined effect of difluoromethylornithine (DFMO) and bcl 2 antisense RNA in the induction of apoptosis of HL60 cells.Methods HL60 cells were transfected with recombinant retroviral vector expressing bcl 2 antisense RNA. The growth characteristics of transfectants were studied by morphological observation, growth curve, FCM analysis, colony formation in soft agar, DNA electrophoresis, molecular hybridization and immunocytochemical staining.Results The recombinant retroviral vectors expressing bcl 2 antisense RNA and the cell line producing recombinant virus were constructed. Although the expression of bcl 2 at mRNA and protein levels in the HL60 cells transfected with antisense bcl 2 were down regulated, there was no change in cell growth, cell cycles and expression of ornithine decarboxylase (ODC) compared with the parental cells albeit some decrease in colony formation. In addition, transfection with antisense bcl 2 did not induce marked apoptosis whereas treatment of the transfectant with low concentration (0.2 mmol/L) of DFMO resulted in enhanced expression inhibition of bcl 2 protein, inhibition of cell growth and induction of apoptosis.Conclusion DFMO synergizes with antiense bcl 2 RNA in the induction of apoptosis of HL60 cells. Subject
出处
《中华肿瘤杂志》
CAS
CSCD
北大核心
2000年第2期105-108,共4页
Chinese Journal of Oncology
