摘要
目的观察RNA干扰技术对肝癌细胞株内骨桥蛋白（osteopontin，OPN）不同片段的抑制效果，以期为针对OPN的靶向治疗提供更准确、有效的位点。方法应用合成OPN序列特异性双链RNA（OPNi-A、OPNiB、OPNi-C），转染人HEP-G2肝癌细胞株，用荧光定量PCR和免疫组化检测比较OPN的mRNA和蛋白表达情况。结果OPNi—A、OPNi—B、OPNiC转染HEP—G2后，A片段的荧光强度大于B片段和C片段。RNA干扰HEP-G2细胞株48h后，A、B、C、三个片段的OPNmRNA的△CT值分别从干扰前的8．31±1.58、8．78±1．49、8．25±1．51上升至12．14±1．43、10．22±1．97、10．48±1．88（P〈0．05）；三个片段的OPN蛋白水平免疫组化评分也从干扰前的6．44±1．67、5．43±2．05、5．45±2．52下降到2．84±1．52、4．43±1．65、3．95±1．43。结论RNA干扰对肝癌细胞株内骨桥蛋白不同片段有不同的抑制作用。合成更具针对性的siRNA可能对抑制肝癌侵袭转移具有重要的科学和卫生经济学意义。
Objective Within human hepatoma cell lines, we aimed to investigate the effects of the down-regulation by RNAi on different fragments of osteopontin （OPN） in order to discover more effective and accurate sites for OPN. Methods Specific small interfering RNA of OPN （OPNi 1） were synthesized and transfected into human hepatoma cell line （HEP G2）. Fluorescent quantitative PCR and immunohistochemical methods were used to test the OPN expression levels of mRNA and protein before and after RNAi. Results After transfection, the ACT value of the A fragment was greater than B and C fragments of OPN mRNA in HEP-G2. Before RNAi was added to HEP-G2 cells, the three fragments A, B, C had OPN mRNA CT values of 8.31＋1.58,8.78±1.49,8.25＋1.51 respectively. Once the RNAi were added, the CT values were measured 48h after for the fragments A, B, and C which were 12.14±1.43,10.22±1.97,10.48±1.88 （P〈0.05） respectively. The immunohistochemical values of A, B, C were down from 6.44±1.67,5.43±2.05,5.45±2.52 to 2.84±1. 52, 4.43± 1.65,3.95± 1.43 respectively after interference. Conclusions RNAi can inhibit the expression of OPN gene selectively, siRNA targets different segments of OPN, which may have more effects on invasion and metastasis of liver cancer for a more important significance in science and health econom itS.
出处
《中华肝胆外科杂志》
CAS
CSCD
北大核心
2012年第4期288-291,共4页
Chinese Journal of Hepatobiliary Surgery
基金
基金项目：广东省科技计划资助项目（2010B031600007）
关键词
骨桥蛋白片段
肝细胞癌
RNA干扰
Osteopontin segment
Hepatacellular carcinoma
RNA interference
