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目的观察硫代修饰型及天然型PCNA反义寡核苷酸在脂质体介导下或直接转染人膀胱癌细胞BIU-87后在细胞内的分布及其稳定性。方法将异疏氰酸荧光素（5＇－FITC）标记的18mer硫代磷酸化修饰型及未修饰型PCNA反义寡核苷酸在脂质体介导下或直接转染人膀胱癌细胞BIU－87，应用荧光显微镜观察转染细胞从细胞内的时相分布。结果修饰型反义核酸直接转染细胞后30分钟，少数细胞胞浆荧光呈离散型、点状分布，4小时后有少数细胞胞核有强荧光聚积。在脂质体介导下，荧光细胞数目明显增加，4小时后绝大多数细胞迅速积聚于细胞核，4～8小时后，细胞核内荧光强度进一步增强，12小时后细胞荧光减弱甚至消失。而非修饰型反义核酸在直接转染或脂质体介导下均发现荧光在3小时后消失。结论脂质体增加PCNA修饰型反义寡核苷酸与膀胱癌细胞BIU－87结合的数量，同时使其在细胞核内分布聚积明显；PCNA反义寡核苷酸硫代修饰具有更好的稳定性。
Objective To observe the distribution and stability of thio-modified and natural PCNA antisense oligonucleotides in the cells after liposome-mediated or direct transfection of human bladder cancer cells BIU-87. Methods The 18mer phosphorothioate-modified and unmodified PCNA antisense oligonucleotides labeled with 5-FITC were transfected into human bladder cancer cells BIU -87, using fluorescence microscopy observed transfected cells from the intracellular phase distribution. Results After 30 days of modified antisense RNA directly transfected into cells, the fluorescence of few cytoplasm showed discrete and dotted distribution. After 4 hours, a few of the cells had strong fluorescence accumulation. Under the condition of liposome, the number of fluorescent cells increased obviously. After 4 hours, the majority of cells rapidly accumulated in the nucleus. After 4 ~ 8 hours, the fluorescence intensity in nucleus was further enhanced, and the fluorescence decreased or even disappeared after 12 hours. The non-modified antisense nucleic acids were found to disappear after 3 hours by direct transfection or liposome-mediated fluorescence. Conclusion Liposomes increase the number of PCNA-modified antisense oligonucleotides binding to bladder cancer cell line BIU-87, and at the same time, they have obvious accumulation and distribution in the nucleus. The antisense oligonucleotide of PCNA has better stability Sex.