siRNA抑制CDK6表达影响鼻咽癌细胞增殖和细胞周期进展

发布时间：2018-01-27 11:15

本文关键词： 鼻咽癌 CDK siRNA 细胞增殖细胞周期　出处：《南方医科大学学报》2014年07期 　论文类型：期刊论文

【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。
[Abstract]:Objective to investigate the effect and molecular basis of siRNA on the growth and cell cycle of nasopharyngeal carcinoma cells after inhibiting CDK6 expression. Methods Real-time fluorescence quantitative PCR was used to detect the expression of CDK6 in 30 cases of nasopharyngeal carcinoma and 18 cases of nasopharynx. The difference of expression. SiRNA was transfected into nasopharyngeal carcinoma (NPC) cell line CNE2. MTT and flow cytometry were used to detect cell proliferation and cell cycle change. The expression of cell cycle related factors was detected by blot. The results were compared with those of nasopharyngeal tissue. The expression of CDK6 gene mRNA in nasopharyngeal carcinoma tissues was significantly increased. After targeting CDK6 expression in nasopharyngeal carcinoma cells, the cell growth ability was significantly decreased. The expression of CDK4CCND1 and E2F1 decreased significantly after inhibiting the expression of CDK6. Conclusion the expression of CDK6 in nasopharyngeal carcinoma is increased. SiRNA-targeted inhibition of CDK6 is followed by down-regulation of CDK4. CCND1 and E2F1 and up-regulation of p21 expression could significantly inhibit the proliferation and cell cycle transformation of nasopharyngeal carcinoma cells.
【作者单位】：右江民族医学院附属医院耳鼻喉科;南方医科大学南方医院影像中心;右江民族医学院附属医院病理科;
【基金】：广西自然科学基金(2013GXNSFBA019193)
【分类号】：R739.63
【正文快照】： 鼻咽癌是发生于鼻咽的一种上皮性恶性肿瘤,好发于我国南方省份以及东南亚等国家和地区,其发病机理与人群易感、EB病毒感染以及环境致癌物等因素相关。许多鼻咽癌相关基因在某种或者多种因素联合作用下异常的激活,促使鼻咽癌发病[1-4]。CDK6是促进细胞周期进展的关键基因之一。

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