彭先兵,戴润芝
PENG Xianbing,DAI Runzhi

• 1:湖北医药学院附属人民医院

摘要(Abstract)：

目的:观察CDK8的异常表达对人鼻咽癌细胞增殖、凋亡及化疗敏感性的影响机制。方法:培养细胞分为未处理组(不给予任何干预)、阴性对照组(不含CDK8表达的慢病毒转染)和转染组(含有CDK8过表达的慢病毒),每组12个培养皿;采用脂质体Lipofectamine TM 2000对鼻咽癌5-8F细胞进行转染,Western-blot法验证转染效率并检测CDK8对相关蛋白的表达,采用MTT和流式细胞术对CDK8对鼻咽癌5-8F细胞增殖、凋亡和细胞周期的进行分析,检测CDK8对化疗药物敏感性的影响。结果:转染pc DNA3.1/CDK8的鼻咽癌5-8F细胞中的CDK8表达明显上调(P<0.05),CDK8异常表达的鼻咽癌5-8F细胞导致增殖能力变慢、导致其凋亡率升高,增殖细胞核抗原(PCNA)蛋白表达水平降低;下调Cyclin D1表达水平(P<0.05);上调细胞凋亡率(P<0.05);多药耐药相关蛋白1(MRP1)表达水平降低(P<0.05); Bcl-2表达水平显著下调(P<0.05); Bax表达水平上调(P<0.05);同时鼻咽癌5-8F细胞能够增加顺铂敏感性(P<0.05)。结论:CDK8的异常表达能够抑制鼻咽癌细胞增殖,上调化疗敏感性;其机制与上调Bax表达、下调Bcl-2、MRP1、PCNA和Cyclin D1表达有关。
Objective:To study the mechanism of abnormal expression of CDK8 on the cell proliferation,apoptosis and chemosensitivity of human nasopharyngeal carcinoma(NPC).Methods:Cultured cells were divided into three groups:the untreated group(without any intervention),the negative control group(without lentivirus transfection with CDK8 expression) and the transfected group(with lentivirus with CDK8 overexpression),with 12 Petri dishes in each group.Liposomal LipofectamineTM2000 was used to transfect NPC 5-8F cells.The transfection efficiency was verified by Western-blot.The proliferation,apoptosis and cycles of NPC 5-8F cells were analyzed by MTT and flow cytometry(CDK8).Western-blot was used to detect the expression of CDK8 related proteins.The sensitivity of CDK8 to chemotherapeutic agents was detected by MTT.Results:The expression of CDK8 in NPC 5-8F cells transfected with pcDNA3.1/CDK8 was up-regulated(P<0.05).This indicates that NPC 5-8F cells with stable expression of CDK8 gene were successfully constructed.The abnormal expression of CDK8 in NPC 5-8F cells led to slow proliferation and increase the rate of apoptosis,leading to a significant decrease in the expression of proliferating cell nuclear antigen(PCNA) protein,and the expression level of Cyclin D1 was significantly reduced(P<0.05).The apoptosis rate significantly increased(P<0.05).The expression level of multidrug resistance associated protein 1(MRP1) significantly decreased(P<0.05).The expression level of Bcl-2 was significantly down regulated(P <0.05),and the expression level of Bax significantly increased(P<0.05).Meanwhile,NPC 5-8F cells significantly increased cisplatin sensitivity(P<0.05).Conclusion: Abnormal expression of CDK8 can inhibit the proliferation of NPC 5-8F cells and increase the chemosensitivity of cancer cells.It is presumed that the mechanism is related to the down regulating expression of Cyclin D1,PCNA,MRP1 and Bcl-2,and the up-regulate expression of Bax.

关键词(KeyWords)： 紫杉醇;鼻咽肿瘤;细胞凋亡;细胞增殖
paclitaxel;nasopharyngeal carcinoma;apoptosis;cell proliferation

Abstract:

Keywords:

基金项目(Foundation):

作者(Author): 彭先兵,戴润芝
PENG Xianbing,DAI Runzhi

DOI: 10.19367/j.cnki.1000-2707.2019.01.024

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