摘要
目的利用siRNA抑制CDK6表达后对鼻咽癌细胞生长及细胞周期的影响及分子基础。方法实时荧光定量PCR检测CDK6在30例鼻咽癌与18例鼻咽组织中的表达差异。siRNA转染鼻咽癌细胞CNE2,靶向抑制CDK6表达。利用MTT和流式细胞仪分别检测细胞增殖和细胞周期改变。Western blot检测细胞周期相关因子的表达改变。结果与鼻咽组织相比,CDK6基因mRNA在鼻咽癌组织中表达明显增高。siRNA在鼻咽癌细胞中靶向抑制CDK6表达后,细胞生长能力明显降低,且细胞周期也明显阻滞在G1期,S期细胞减少。机制分析显示在抑制CDK6表达后,细胞周期促进因子CDK4、CCND1和E2F1表达显著下降,而细胞周期抑制因子p21表达明显提升。结论 CDK6在鼻咽癌中表达升高。siRNA靶向抑制CDK6之后通过下调CDK4、CCND1和E2F1以及上调p21表达后,能明显抑制鼻咽癌细胞增殖能力及细胞周期转化。
Objective To assess the effect of small interfering RNA （siRNA）-mediated suppression of CDK6 expression on the proliferation and cell cycles of nasopharyngeal carcinoma （NPC） cells in vitro. Methods QRT-PCR was used to examine the differential expression of CDK6 in 30 NPC tissues and 18 normal nasopharyngeal tissues. A siRNA targeting CDK6 was transfected in NPC CNE2 cells, and MTT assay and flow cytometry were used to analyze the changes in cell proliferation and cell cycle distribution. Western blotting was used to examine the expressions of the cell cycle-related factors. Results Compared with normal nasopharyngeal tissues, NPC tissues showed an increased expression of CDK6 mRNA. Knocking down CDK6 expression obviously inhibited tumor cell growth and cell cycle transition from G1 to S phase and caused reduced expressions of CDK4, CCND1, and E2F1 and enhanced expression of the tumor suppressor p21. Conclusion NPC tissues overexpress CDK6. Knocking down CDK6 expression inhibits the growth and cell cycle transition of NPC cells in vitro by inhibiting the expressions of CDK4, CCND1, and E2F1 and upregulating tumor suppressor p21 expression.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2014年第7期1071-1074,共4页
Journal of Southern Medical University
基金
广西自然科学基金(2013GXNSFBA019193)
关键词
鼻咽癌
CDK6
SIRNA
细胞增殖
细胞周期
nasopharyngeal carcinoma
CDK6
small interfering RNA
cell proliferation
cell cycle
