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首页> 中文期刊> 《临床肿瘤学杂志》 >转录水平RNA干扰肝素酶基因对肝癌细胞侵袭能力的影响

转录水平RNA干扰肝素酶基因对肝癌细胞侵袭能力的影响

         
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Objective To study the effects of transcription gene sciencing ( TGS ) and post-transcriptional gene silencing ( PTGS) on interfernce of heparanase( HPA) gene in hepatocellular carcinoma cell line SMMC-7721 and its invasion ability. Methods The small interfering( siRNA) of TGS and PTGS covered promoter region and coding region of HPA gene were designed and synthe-ses, and then transfected into hepatocellular carcinoma cell line SMMC-7721 by neuclear trancfection. HPA expression in both mRNA and protein level were detected by RT-PCR and Western blotting at 48, 72 and 96h, and blank group was set as control. Transwell chamber experiment was used to detect the ability of invasion and migration of hepatocellular carcinoma cell line SMMC-7721 after TGS and PTGS siRNA transfected. Results Both of TGS and PTGS technique could silence HPA gene at mRNA and protein level 48 hours later after siRNA transfection. The expression of HPA gene in PTGS group was recovered 72 hours later after siRNA transfection, but HPA still kept silencing in TGS group. However, the expression of HPA gene in both of TGS and PTGS groups were recovered 96 hours later after gene transfection. The transwell chamber experiment confirmed that TGS and PTGS of HPA gene could reduce the abilities of invasion of SMCC-7721 cell, especially in TGS group. Conclusion TGS can silence HPA gene better compared with PTGS,and sig-nificantly decrease the abilities of invasion of hepatocellular carcinoma cell line SMMC-7721.%目的：探讨转录水平基因沉默（ TGS）技术和转录后水平基因沉默（ PTGS）技术对肝素酶（ HPA）基因干扰效果及其对肝癌SMCC-7721细胞侵袭能力的影响。方法从HPA基因启动子区和编码区分别设计并合成TGS和PTGS的小干扰RNA（ siRNA），并转染肝癌细胞SMMC-7721；实时半定量PCR和Western blotting检测TGS和PTGS siRNA转染后48、72和96h HPA的表达，并设空白组作为对照；Transwell小室实验检测干扰后SMMC-7721细胞的侵袭能力。结果 TGS和PTGS两种技术在转染48h后，从mRNA和蛋白水平上均能成功干扰HPA表达；转染后72h，PTGS组HPA恢复表达，而TGS组仍保持沉默；转染后96h，两组HPA均恢复表达。 Transwell小室实验表明，TGS和PTGS组HPA基因均能使SMCC-7721的穿膜细胞数减少，TGS组效果更明显。结论 TGS技术沉默肝癌SMMC-7721细胞中HPA基因的能力优于PTGS技术，并使肝癌细胞的侵袭能力显著降低。

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