摘要
目的:研究地西他滨(DAC)对体外培养的人鳞状上皮舌癌SCC-9细胞p16基因甲基化状态及m RNA和蛋白表达的影响。方法:实验将体外培养的人鳞状上皮舌癌SCC-9细胞分为0、1、2、3四组。1、2、3三组样本采用3个梯度浓度的DAC进行处理,取未经DAC处理的0组作为对照组。药物处理48 h后,Q-MSP检测SCC-9细胞p16基因甲基化状态;q PCR技术检测SCC-9细胞p16基因m RNA的表达;免疫组化SP法检测SCC-9细胞p16蛋白表达。结果:Q-MSP检测结果显示SCC-9细胞p16基因甲基化与非甲基化呈杂合状态。Real-time PCR结果显示实验组SCC-9细胞p16基因m RNA与对照组相比表达上调(P<0.05)。免疫组化结果显示实验组中p16蛋白较对照组呈现高表达状态。结论:DAC可能具有逆转SCC-9细胞p16基因甲基化状态,并具有促使p16基因m RNA和蛋白恢复表达的作用。
Objective To study the effects of deoxycytidine(5-aza-2 deoxycytidine, DAC) on DNA Methylation state and expression of m RNA and protein of pl6 gene in human squamous lingual carcinoma SCC-9cells in vitro. Methods The SCC-9 cells were divided into four groups, group 0, 1, 2 and 3 which processed using three gradients concentration of DAC. The group 0 without DAC was as the control group. Q-MSP was used to detect the state of methylation of the p16 in SCC-9 cells treated by DAC after 48 hours. Real-time fluorescence quantitative PCR was used to detect m RNA expression level changing of the p16 in SCC-9 cells treated by DAC.Immunohistochemical method was used to detect the expression of p16 protein. Results The hypermethylation and non-methylated p16 gene in SCC-9 was mixed with the results of Q-MSP. The results of Real-time PCR showed that m RNA expression of p16 in SCC-9 cells which treated by the different concentration of DAC for 48 hours was higher the control group. And difference was statistically significant(P 0.05). The high expression of p16 protein was found in the experimental group with immunohistochemical method. Conclusion The p16 gene methylation states of SCC-9 may be suppresses and the recovery of m RNA and protein expression of p16 gene must be prompted by DAC.
出处
《实用医学杂志》
CAS
北大核心
2016年第16期2613-2616,共4页
The Journal of Practical Medicine
基金
广西高校科学技术研究资助项目(编号:ZD2014035)
广西医疗卫生重点科研课题(编号:重2011033)
2015广西科学研究与技术开发计划(编号:12-18)