磁共振成像评价腺病毒介导的酪氨酸酶基因在HepG2细胞的表达  

Adenovirus Mediated Gene Transfer of Tyrosinase Gene on HepG2 Cell by Magnetic Resonance Imaging

作　　者：元建鹏[1] 梁碧玲[1] 邓贺然[1] 刘壮盛[1] 白守民[1] 钟镜联[1] 

机构地区：[1]中山大学附属第二医院放射科,广州510120

出　　处：《中国医学科学院学报》2009年第2期146-150,共5页Acta Academiae Medicinae Sinicae

摘　　要：目的以酪氨酸酶基因腺病毒重组体转染HepG2细胞,以磁共振成像(MRI)观察酪氨酸酶基因腺病毒重组体转染体外细胞的效果。方法以不同数量酪氨酸酶基因腺病毒重组体转染HepG2细胞,以MRIT1加权像(T1WI)、T2加权像(T2WI)及短时间间隔反转恢复序列(STIR)扫描转染细胞。应用Masson-Fontana染色检测黑色素的合成,实时定量PCR验证酪氨酸酶基因的转染与表达。结果酪氨酸酶基因腺病毒重组体转染HepG2细胞并在其中表达生成黑色素,经转染复数分别为50、150、300的重组腺病毒转染的1×106个细胞内生成的黑色素能够被MRI检测到并在MRIT1WI、T2WI、STIR检查呈高信号。Masson-Fontana染色检测到转染的HepG2细胞内的黑色素颗粒;实时定量PCR在转染细胞中检测到的酪氨酸酶基因的cDNA表达量较未转染细胞的表达量明显增高。结论MRI能够检测到HepG2细胞由外源基因表达合成的黑色素,表明腺病毒作为运送载体可以有效地运送酪氨酸酶基因进入HepG2细胞。Objective To evaluate the transfect results of recombinant adenovirus vector carrying tyrosinase gene （Ad-tyr） in vitro by magnetic resonance imaging （MRI） after the Ad-tyr was transfected into HepG2 cell. Methods The Ad-tyr which carried the full-length cDNA of tyrosinase gene was transfected into HepG2 cell. The transfected cells were scan by MRI sequences of T1 weighted image （T1WI） , T2 weighted image （T2WI） , and short time inversion recovery （STIR） to observe the MRI signals of expressed melanin. Masson-Fontana staining was performed to search for melanin granules in transfected cells. Real-time PCR method was used to search for cDNA of tyrosinase gene. Results Ad-tyr was transfected into HepG2 cells and synthesized a large amount of melanin inside. The synthesized melanin of 1 × 10^6 cells which had been transletted by Ad-tyr with the 50, 150, and 300 multiplicity of infection separately were all sufficient to be detected by MRI and showed high signals in MRI T1WI, T2WI, and STIR sequences. The signal intensities of MRI were positively correlated to the amounts of transfected Ad-tyr. The melanin granules were found in HepG2 cells in Masson-Fontana staining. The cDNA amount of tyrosinase gene in transfected HepG2 cells, which was detected by real-time PCR, was remarkably higher than that in nontransfected cells. Conclusion The synthesized melanin of HepG2 cells, which controlled by expression of exogenous gene, can be detected by MRI, indicating that the adenovirus vector can efficiently carry the tyrosinase gene into HepG2 cells.

关 键 词：酪氨酸酶基因 腺病毒 磁共振成像 

分 类 号：R445.2[医药卫生—影像医学与核医学]