CAJ | 학술논문

目的：研究运用 RNA 干扰（RNA interference，RNAi）技术抑制 NF -κB/ p65基因的表达对人肺腺癌细胞株 A5492细胞增殖、周期及凋亡的影响，探讨 NF -κB/ p65基因在肺腺癌发生发展中的作用。方法：利用阳离子脂质体 LipofectamineTM2000将人 NF -κB/ p65的小干扰 RNA 转染入肺腺癌细胞株 A549中。运用 real time - PCR 和 Western blot 技术检测 A549细胞内 NF -κB/ p65、增殖相关基因 Cyclin D1和凋亡相关基因 Bcl-2、Bax 的 mRNA 的转录水平和蛋白的表达情况，MTT 法检测细胞的增殖活力，显微镜下观察细胞的生长情况。结果：NF -κB/ p65 siRNA 能有效地抑制 A549细胞中 NF -κB / p65基因在 mRNA 水平上的表达（P﹤0.001），同时下调 Cyclin D1和 Bcl -2的表达（均 P ﹤0.01）。上调 Bax 表达（P ﹤0.001）；蛋白表达也具同样趋势。MTT 实验证明 p65 siRNA 转染组中细胞增殖减慢；显微镜镜下观察显示，细胞生长减慢，凋亡形态改变明显。结论：体外实验初步证明 NF -κB/ p65基因在肺腺癌细胞株 A549增殖分化与凋亡方面扮演重要角色，通过沉默其表达可抑制肺腺癌细胞株 A549增殖并诱导其凋亡。进一步证实 NF -κB/ p65对 Cyclin D1、Bcl -2、Bax 具有调控作用，他们共同参与了肺腺癌的发生发展过程。
목적：연구운용 RNA 간우（RNA interference，RNAi）기술억제 NF -κB/ p65기인적표체대인폐선암세포주 A5492세포증식、주기급조망적영향，탐토 NF -κB/ p65기인재폐선암발생발전중적작용。방법：이용양리자지질체 LipofectamineTM2000장인 NF -κB/ p65적소간우 RNA 전염입폐선암세포주 A549중。운용 real time - PCR 화 Western blot 기술검측 A549세포내 NF -κB/ p65、증식상관기인 Cyclin D1화조망상관기인 Bcl-2、Bax 적 mRNA 적전록수평화단백적표체정황，MTT 법검측세포적증식활력，현미경하관찰세포적생장정황。결과：NF -κB/ p65 siRNA 능유효지억제 A549세포중 NF -κB / p65기인재 mRNA 수평상적표체（P﹤0.001），동시하조 Cyclin D1화 Bcl -2적표체（균 P ﹤0.01）。상조 Bax 표체（P ﹤0.001）；단백표체야구동양추세。MTT 실험증명 p65 siRNA 전염조중세포증식감만；현미경경하관찰현시，세포생장감만，조망형태개변명현。결론：체외실험초보증명 NF -κB/ p65기인재폐선암세포주 A549증식분화여조망방면분연중요각색，통과침묵기표체가억제폐선암세포주 A549증식병유도기조망。진일보증실 NF -κB/ p65대 Cyclin D1、Bcl -2、Bax 구유조공작용，타문공동삼여료폐선암적발생발전과정。
To study the effect of suppression of NF - κB/ p65 gene expression on proliferation and ap-optosis of human lung adenoma cell line A549,and to explore the role of NF - κB/ p65 in generation and development of human lung adenoma. Methods:Small interference RNA(siRNA)directed against human NF - κB/ p65 was trans-fected into human lung adenoma A549 cells by using cationic liposome LipofectamineTM2000. The expression of NF -κB/ p65 and proliferation - related gene Cyclin D1 gene and apoptosis - related genes Bcl - 2 and Bax were detected by using real - time PCR. The effect of cell proliferation was studied by MTT. Morphological changes of the cell were observed by optical microscope. Results:The results of real - time PCR indicated that siRNA could knock down the transcription and expression of NF - κB/ p65(P ﹤ 0. 001). The expressions of Cyclin D1 and Bcl - 2 mRNA as well as their proteins in p65 siRNA group were significantly downregulated,but the expression of Bax mRNA was markedly upregulated compared to non - transfected cells. There were significant differences among three groups(P ﹤ 0. 01). The inhibitory action on proliferation of p65 siRNA group cells was confirmed by MTT. The apoptotic rate of p65 siR-NA group was significantly higher than control groups. Conclusion:The study provided basis for researching the func-tion of NF - κB/ p65 gene,indicating the expression of NF - κB down - regulating might inhibit proliferation and in-duce apoptosis in human lung adenoma A549 cells via regulation of the expression of Cyclin D1,Bcl - 2 and Bax.