中华肝胆外科杂志
中華肝膽外科雜誌
중화간담외과잡지
CHINESE JOURNAL OF HEPATOBILIARY SURGERY
2012年
4期
288-291
,共4页
林帆%古维立%范少峰%李坤平%林春明%裴正浩
林帆%古維立%範少峰%李坤平%林春明%裴正浩
림범%고유립%범소봉%리곤평%림춘명%배정호
骨桥蛋白片段%肝细胞癌%RNA干扰
骨橋蛋白片段%肝細胞癌%RNA榦擾
골교단백편단%간세포암%RNA간우
Osteopontin segment%Hepatacellular carcinoma%RNA interference
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、OPNi-B、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、OPNi-C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48 h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.
目的 觀察RNA榦擾技術對肝癌細胞株內骨橋蛋白(osteopontin,OPN)不同片段的抑製效果,以期為針對OPN的靶嚮治療提供更準確、有效的位點.方法 應用閤成OPN序列特異性雙鏈RNA(OPNi-A、OPNi-B、OPNi-C),轉染人HEP G2肝癌細胞株,用熒光定量PCR和免疫組化檢測比較OPN的mRNA和蛋白錶達情況.結果 OPNi-A、OPNi-B、OPNi-C轉染HEP-G2後,A片段的熒光彊度大于B片段和C片段.RNA榦擾HEP-G2細胞株48 h後,A、B、C、三箇片段的OPNmRNA的△CT值分彆從榦擾前的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)；三箇片段的OPN蛋白水平免疫組化評分也從榦擾前的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.結論 RNA榦擾對肝癌細胞株內骨橋蛋白不同片段有不同的抑製作用.閤成更具針對性的siRNA可能對抑製肝癌侵襲轉移具有重要的科學和衛生經濟學意義.
목적 관찰RNA간우기술대간암세포주내골교단백(osteopontin,OPN)불동편단적억제효과,이기위침대OPN적파향치료제공경준학、유효적위점.방법 응용합성OPN서렬특이성쌍련RNA(OPNi-A、OPNi-B、OPNi-C),전염인HEP G2간암세포주,용형광정량PCR화면역조화검측비교OPN적mRNA화단백표체정황.결과 OPNi-A、OPNi-B、OPNi-C전염HEP-G2후,A편단적형광강도대우B편단화C편단.RNA간우HEP-G2세포주48 h후,A、B、C、삼개편단적OPNmRNA적△CT치분별종간우전적8.31±1.58、8.78±1.49、8.25±1.51상승지12.14±1.43、10.22±1.97、10.48±1.88 (P＜0.05)；삼개편단적OPN단백수평면역조화평분야종간우전적6.44±1.67、5.43±2.05、5.45±2.52하강도2.84±1.52、4.43±1.65、3.95±1.43.결론 RNA간우대간암세포주내골교단백불동편단유불동적억제작용.합성경구침대성적siRNA가능대억제간암침습전이구유중요적과학화위생경제학의의.
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 △CT 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 immunohis tochemical 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 economics.
