摘要
目的检测PSF1在结肠癌组织中的表达，探讨RNA干扰沉默PSF1表达对人结肠癌细胞增殖的影响及其可能的机制。方法收集2004年5月至2006年12月间经病理确诊的40例结肠癌标本．采用Westerllblot检测标本中PSF1蛋白表达水平；应用脂质体法将靶向PSF1的短发夹RNA（shRNA）干扰质粒转染人结肠癌细胞株LOVO、HT．29和HCT116，Westernblot法检测PSF1蛋白表达变化．分别应用MTY法、软琼脂克隆形成实验及荧光定量PCR检测转染PSF1shRNA干扰质粒对结肠癌细胞增殖活性、锚定非依赖性生长能力及PSF2、PSF3和SLD5mRNA表达的影响。结果结肠癌组织中PSF1蛋白相对表达量为0．485±0．113，显著高于正常黏膜组织的0．056±0．014（P〈0．01）。转染PSF1sbRNA干扰质粒后，LOVO、HT-29和HCT116细胞中PSF1蛋白表达水平均显著下降（P〈0．05），细胞增殖能力显著降低，软琼脂克隆形成率明显下降。与阴性对照组及正常生长组细胞比较，差异均有统计学意义（P〈0．05）。转染PSF1shRNA干扰质粒后．LOVO、HT-29和HCT116细胞中PSF2、PSF3和SLD5mRNA表达均明显下降（P〈0．05）。结论PSF1与结肠癌的发生发展具有一定关系。利用RNA干扰技术能有效抑制结肠癌细胞中PSF1表达。并抑制结肠癌细胞增殖。PSF1基因有望成为结肠癌靶向治疗的新靶点。
Objective To detect the expression of PSFI （partner of Sld five 1 ） in colon cancer specimens, and to explore the effect of RNA interference targeting PSF1 on the proliferation of colon cancer cells and its mechanism. Methods Expression level of PSF1 protein in colon cancer specimens was detected by Western blot in 40 patients with colon cancer from May 2004 to December 2006. The short hairpin RNA（shRNA） plasmid targeting PSF1 was transfected into LOVO, HT-29 and HCTll6 cells with liposome, then the expression level of PSF1 protein was measured by Western blot, the effect of PSF1 shRNA plasmid transfection on cell proliferation by MTY assay, anchorage-independent growth by soft agar colomy-formation assay, and PSF2, PSF3 and SLD5 mRNA expression by quantitative reverse transcription polymerase chain reaction. Results The relative expression level of PSF1 protein in colon cancer tissues was 0.485±0.113, which was significantly higher than that in adjacent normal mucosa tissues （0.056±0.014, P〈0.01）. Western blot showed that the expression level of PSF1 protein was significantly decreased in colon cancer cells transfected with PSF1 shRNA plasmid. After PSF1 shRNA plasmid transfection, cell proliferation was significantly suppressed, the soft agar colony-forming rates of LOVO, HT-29 and HCTll6 cells were significantly lower than those in control groups （P〈0.05）, meanwhile the expression levels of PSF2, PSF3 and SLD5 mRNA were significantly decreased （P〈0.05）. Conclusions PSF1 is significantly up-regulated in colon cancer tissues compared with adjacent normal mucosa tissues. ShRNA plasmid targeting PSF1 can inhibit the expression of PSFI gene, suppress the proliferation of colon cancer ceils, suggesting that it may be a new therapeutic target for colon cancer.
出处
《中华胃肠外科杂志》
CAS
CSCD
2013年第1期70-74,共5页
Chinese Journal of Gastrointestinal Surgery
基金
国家自然科学基金（30872462）
广东省自然科学基金重点项目（10251008901000011）
