^(31)P-MRS检测标记基因表达肝直接基因转导初步研究

Preliminary study on monitoring marked gene expression of liver direct gene transduction by ^(31)P-MRS

目的:探讨31P-MRS技术在兔VX2肝移植瘤模型经AdCMVIL12-IRES-CKb腺病毒基因转导后通过标记基因间接表达的可行性。方法:健康新西兰大白兔20只,采用开腹包埋法建立兔VX2肝肿瘤模型,将种植成功的18只实验兔随机分成两组,于3周左右行31P-MRS扫描后采用开腹法分别将等量AdCMVIL12-IRES-CKb腺病毒和生理盐水注射到两组瘤兔肿瘤及周围组织中,肌酸水溶液饲养5天后再次行31P-MRS扫描。同时通过免疫组化、琼脂糖凝胶蛋白印迹法(Western Blot)对IL-12和CK进行检测。结果:经AdCMVIL12-IRES-CKb腺病毒治疗后的兔肝VX2肿瘤组织PCr峰较对照组明显升高,分别为(0.84±0.41)、(0.23±0.29)mmol/L,两组治疗前后PCr比较,P值小于0.05,差异有统计学意义;免疫组化在治疗组肝脏局部组织中检测到IL-12的表达,对照组未见表达;Western Blot法检测到治疗组CKB的表达,但对照组未见表达。结论:通过31P-MRS成像检测病毒直接基因转导后体内肝脏的基因表达是可行的。

Objective：To explore the feasibility of ^31P-MRS technology expressing indirectly by marker gene in the rabbit VX2 liver transplantation tumor models after AdCMVIL12-IRES-CKb adenoviral gene transducting.Methods：20 healthy New Zealand white rabbits were used to establish the rabbit VX2 hepatic tumor models by sigmoidectomy embedding method.18 experimental rabbits successfully planted were divided into two groups at random and were scanned by^ 31P-MRS about three weeks,and then respectively injected AdCMVIL12-IRES-CKb adenovirus and physiological saline into tumor and surrounding tissues by sigmoidectomy embedding method.^31P-MRS scan was again performed after animals were fed with creatine solution for five days.Meanwhile,IL-12 and CK were detected by immunohistochemistry（IHC） staining and agarose gel protein imprinting method（western blot）.Results：After treatment of AdCMVIL12-IRES-CKb adenovirus,PCr peak in rabbits liver tumor tissues was obviously higher than that in control group,（0.84±0.41）,（0.23±0.29） mmol/L respectively;pre-and post-treatment among the two groups,PCr peak is statistically significant（P〈0.05）;the expressions of IL12 and CKB were detected in the liver in the experimental group by immunohistochemistry（IHC） staining and Western blot respectively,but not shown in the control group.Conclusion：It is feasibility to indirectly detect gene expression in liver by phosphorous-31 magnetic resonance spectroscopy（^31P-MRS） after virus directly gene transduction.