| 王国栋 党亚正 魏铭 郑甲 温伟红.siRNA下调EZH2 基因对肾癌细胞系769-P增殖的影响[J].,2014,14(23):4440-4443 |
| siRNA下调EZH2 基因对肾癌细胞系769-P增殖的影响 |
| Effect of EZH2 after Silenced by siRNA on the Proliferation of Renal Cell Carcinoma Cell Line 769-P |
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| DOI： |
| 中文关键词: 肾细胞癌 RNAi EZH2 |
| 英文关键词: Renal cell carcinoma RNAi EZH2 |
| 基金项目:国家自然科学基金面上项目(81171924) |
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| 中文摘要: |
| 目的：运用小干扰RNA下调果蝇zeste 基因增强子人类同源物（enhancer of zeste homolog 2，EZH2）在肾癌细胞系769-P 中
的表达，明确其对肾癌细胞增殖的影响。方法：将处于对数生长期769-P 细胞分为实验组（experiment group）、阴性对照组(negative
group)、空白对照组（blank group），合成靶向EZH2 基因的小干扰RNA片段（EZH2-siRNA）和无效序列片段后，通过脂质体介导分
别转染至实理组和阴性对照组，空白对照组未做任何处理。以qRealtime-PCR 检测EZH2 基因mRNA 水平的变化情况，以MTT
法检测各组细胞增殖变化；流式细胞术（FCM）检测转染后细胞周期变化情况。结果：实理组中EZH2 在mRNA 表达水平明显受
抑制；MTT实验中第4 天始，实验组中769-P 细胞的增殖能力开始受抑制，第5 天时实验组细胞抑制更明显，与阴性对照组和空
白组比较差异有统计学意义（P < 0.05）。siRNA 转染后实验组中G0/G1 期细胞比例明显增多(81.32± 3.14)%，与阴性对照组
(44.13± 1.52)%和空白对照组(45.71± 2.32)%差异有统计学意义。结论：EZH2-siRNA 可有效下调并抑制肾癌细胞769-P的增殖，
EZH2在肾癌的发生、发展中发挥了重要作用，为下一步研究肾癌基因治疗提供了理论支持。 |
| 英文摘要: |
| Objective:To investigate the influence of siRNA targeting the enhancer of homolog2(EZH2) gene on the proliferation
of renal cell carcinoma (RCC) cell line 769-P in vitro.Methods:The cell line 769-P which were in the period of logarithmic phase were
divided into experiment group, negative group and blank group. siRNA sequence targeting EZH2 were designed and synthesized
according to genebank, with a scramble sequence as negative control, which were transefected to the 769-P cells conducted by
Lipofectamine 2000. The expression of EZH2 in mRNA level was detected by qRT-PCR after downregulated. MTT assay was used to
observe the growth of 769-P cells in experiment group, negative group and blank group. And, cell cycle distribution of 769-P cells were
detected by flow cytometry.Results:The expression of EZH2 in mRNA level was significantly suppressed by siRNA. MTT assay results
revealed that cell inhibitory rate of experiment group was more lower than that of negative group and blank group. The proliferation of
769-P cells were inhibited remarkably in experiment group with cell rate of G0/G1 phrase being (81.32± 3.14)%, and there were statistic
difference as compared with negative control group (44.13± 1.52)%and blank group(45.71± 2.32)%.Conclusion:EZH2 gene silencing
can be conducted effectively by siRNA. EZH2 may play important roles in the development and progression of RCC, EZH2 may be a
potential theraputic target, and provides an important theoretical basis for gene therapy of RCC. |
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