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目的探讨能否通过siRNA人工诱导RNA干扰,在EB病毒阳性的鼻咽癌细胞株C611中,选择性抑制潜伏膜蛋白1(LMP1)的表达,并了解LMP1抑制对鼻咽癌细胞生长的影响。方法利用脂质体转染的方法,将人工合成的双链siRNA导入鼻咽癌细胞,通过半定量RT-PCR检测LMP1 mRNA水平的变化。并采用MTT法和流式细胞仪检测LMP1表达抑制后,鼻咽癌细胞增殖和细胞周期的变化。结果C611细胞转染siRNA可使LMP1 mRNA水平下降90%以上。LMP1基因抑制后,C611细胞增殖速度下降33%;流式细胞检测显示,C611细胞周期在G0-G1期受阻。结论本研究证明,全新的人工诱导RNA干扰技术,能够有效抑制LMP1基因的表达,并降低鼻咽癌细胞的增殖能力,为鼻咽癌研究和抗肿瘤基因治疗提供了新的思路。
OBJECTIVE: To investigate whether LMP1 can selectively inhibit the expression of latent membrane protein 1 (LMP1) in Epstein-Barr virus-positive nasopharyngeal carcinoma cell line C611 by RNA interference induced by RNAi and to understand the effect of LMP1 inhibition on the growth of nasopharyngeal carcinoma cells. Methods Liposome-mediated transfection of synthetic double-stranded siRNA into nasopharyngeal carcinoma cells was used to detect the level of LMP1 mRNA by semi-quantitative RT-PCR. The proliferation and cell cycle of nasopharyngeal carcinoma cells were detected by MTT assay and flow cytometry after inhibition of LMP1 expression. Results Transfection of siRNA with C611 cells reduced LMP1 mRNA levels by more than 90%. After LMP1 gene inhibition, the proliferation rate of C611 cells decreased by 33%. Flow cytometry showed that the cell cycle of C611 was blocked at G0-G1 phase. Conclusion This study demonstrates that the new artificial RNA interference technology can effectively inhibit the expression of LMP1 gene and reduce the proliferation of nasopharyngeal carcinoma cells, providing a new idea for the study of nasopharyngeal carcinoma and anti-tumor gene therapy.