• The Korean Journal of Nuclear Medicine
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• v.35 no.3
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• pp.168-184
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• 2001

Purpose: KR-30035 (KR), a new MDR reversing agent, has been found to produce a similar degree of increased Tc-99m MIBI uptake in cultured tumor cells over-expressing mdr1 mRNA compared to verapamil (VP), with less cardiovascular effects. We assessed the MDR-reversing ability of KR in vivo, and effects of various doses of KR on MIBI uptake un nude mice hearing P-glycoprotein (P-gp) positive (+) and P-gp negative (-) human tumor xenografts. Methods: P-gp (+) HCT15/CL02 colorectal and P-gp (-) A549 non-small cell cancer cells were inoculated in each flank of 120 nude mice (20 mice ${\times}$ 6 groups). Group 1 (Gr1) mice received 10mg/kg KR i.p. 3 times $({\times}3)$; Gr2, 10mg/kg VP i.p. ${\times}3$; Gr3, 10mg/kg KR i.p. ${\times}2$ + 25mg/kg KR i.p. ${\times}1$; Gr4, 10mg/kg KR i.p. ${\times}2$ + 50mg/kg i.p. ${\times}1$; Gr5, 10mg/kg KR i.p. ${\times}2$ + 25mg/kg KR i.v. ${\times}1$, GrC, controls. The mice were then injected with Tc-99m MIBI and sacrificed after 10 min, 30 min, 90 min and 240 min. Tumor uptake of MIBI (TU) in each group was compared. Results: TU in P-gp (+) and (-) tumors were both higher in Gr1 than Gr2. Washout rate between the 10 min and 4 hours was lower in Gr5 of P-gp (+) cell(0.93) than the control. Percentage increases in TU were higher in P-gp (+) than P-gp (-) tumors with all KR doses. Pgp (+) TU were highest at 10 mon (173% of GrC) and persisted up to 240 min (144%) in Gr3. Larger doses of KR resulted in a lesser degree of increase in P-gp (+) TU at 10 min (130% in Gr4 and 117% un Gr5) and 30 min (178%, 129%), but TU increased by time up to 240 min (177%, 196%). Heart and lung uptakes were markedly increased in Gr4 and Gr5 at 10 and 30 min, likely due to cardiovascular effects. No mice died. Conclusion: These data further suggest that KR that has significantly lower cardiovascular toxicity than verapamil can be used as an active inhibitor of MDR. Even a relatively low dose of KR significantly increased Tc-99m MIBI uptake in P-gp (+) tumors in vivo.

• The Korean Journal of Nuclear Medicine
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• v.37 no.6
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• pp.418-427
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• 2003

Objects: $^{99m}-lactosylated$ human serum albumin (LSA) is a newly synthesized radiopharmaceutical that binds to asialoglycoprotein receptors, which are specifically presented on the hepatocyte membrane. Hepatic uptake and blood clearance of LSA were evaluated in rat with acute hepatic injury induced by dimethylnitrosamine (DMN) and results were compared with corresponding findings of liver enzyme profile and these of histologic changes. Materials and Methods: DMN (27 mg/kg) was injected intraperitoneally in Sprague-Dawley rat to induce acute hepatic injury. At 3(DMN-3), 8(DMN-8), and 21 (DMN-21) days after injection of DMN, LSA injected intravenously, and dynamic images of the liver and heart were recorded for 30 minutes. Time-activity curves of the heart and liver were generated from regions of interest drawn over liver and heart area. Degree of hepatic uptake and blood clearance of LSA were evaluated with visual interpretation and semiquantitative analysis using parameters (receptor index : LHL3 and index of blood clearance : HH3), analysis of time-activity curve was also performed with curve fitting using Prism program. Results: Visual assessment of LSA images revealed decreased hepatic uptake in DMN treated rat, compared to control group. In semiquantitative analysis, LHL3 was significantly lower in DMN treated rat group than control rat group (DMN-3: 0.842, DMN-8: 0.898, DMN-21: 0.91, Control: 0.96, p<0.05), whereas HH3 was significantly higher than control rat group (DMN-3: 0.731,.DMN-8: 0.654, DMN-21: 0.604, Control: 0.473, p<0.05). AST and ALT were significantly higher in DMN-3 group than those of control group. Centrilobular necrosis and infiltration of inflammatory cells were most prominent in DMN-3 group, and were decreased over time. Conclusion: The degree of hepatic uptake of LSA was inversely correlated with liver transaminase and degree of histologic liver injury in rat with acute hepatic injury.

• The Korean Journal of Nuclear Medicine
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• v.4 no.1
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• pp.27-36
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• 1970

적혈구(赤血球) 수명의 측정에는 $^{51}Cr$-표지적혈구법(標識赤血球法)이 임상적(臨床的)으로 이용(利用)되고 있으며 이는 이론상(理論上) steady state 즉(卽) 측정기간(測定期間)동안 순환(循環) $^{51}Cr$량(量)-적혈구량(赤血球量)이 일정(一定)할 때에 한(限)하여 유효(有效)하며 unsteady state 때는 true red cell survival을 알기 위하여서는 측정치에 영향을 주는 요인(要因)에 대하여 각각(各各) 교정(校正)해 줄 필요(必要)가 있다. 이 요인(要因)중에 특히 실혈(失血)로 인(因)한 영향에 관(關)하여는 계통적인 연구(硏究)가 적다. 이에 저자(著者)들은 $^{51}Cr$표지적혈구법(標識赤血球法)을 이용(利用)하여 실혈(失血)이 적혈구(赤血球) 수명측정(測定)에 미치는 영향을 인체(人體)에서 실험 관찰하여 몇가지 성적을 얻었다. 연구대상(硏究對象)은 총(總) 56명(名)의 청장년(靑壯年)으로 급성실혈군(急性失血群)과 만성실혈군(慢性失血群)으로 구분(區分)하여 급성실혈군(急性失血群)은 위장출혈등(胃腸出血等)이 없는 2대(代)의 의대생(醫大生)으로 $^{51}Cr$표지적혈구법(標識赤血球法)을 사용하여 적혈구(赤血球) 수명을 측정하는 동안($10{\sim}14$ 일간(日間)) 1일당(日當) 10ml(6명(名)), 25ml(4명(名)), 50ml(4명(名)), 75ml(4명(名)), 100ml(6명(名))를 각각(各各) 사혈(瀉血)한 군(群)과 10일간(日間) 1,000ml를 사혈한 군(群) 즉 200ml씩 5회(回)(4명(名)), 500ml씩 2회(回)(4명(名))로 세분(細分)하였으며 만성실혈군(慢性失血群)은 직업적인 공혈자(供血者)로 반복사혈로 생긴 9명(名)의 빈혈자와 십이지장충증(十二指腸蟲症)에 감염(感染)되어 구충(驅蟲)한 중등도(中等度)의 철결핍성 빈혈환자 7명(名)으로 나누어 관찰하였다. 측정(測定) 방법(方法)으로는 Gray 및 Sterling법(法)을 개설한 방법(方法)으로 $^{51}Cr$표지적혈구(標識赤血球)의 계측시료(計測試料)로서 전혈(全血) 및 적혈구(赤血球)를 사용(使用)하였다. 실험(實驗)성적은 1. 1일당(日當) 실혈량(失血量)이 증가(增加)할수록 적혈구(赤血球)수명($T\frac{1}{2}$)은 짧아짐을 알 수 있었다. 즉(卽) 1일당(日當) $20{\sim}50ml$ 사혈군에서는 $T\frac{1}{2}$이 현저히 짧아지는 rapid phase을 나타내고 1일당(日當) 50ml이상(以上) 사혈군에서는 짧아지는 정도(程度)가 완만한 slow phase을 나타낸다(Fig. 6). 2. 1일량(日量) 10ml 및 25ml식(式) 사혈한 군(群)의 적혈구수명(赤血球壽命)을 측정(測定)하는데 있어 적혈구(赤血球)를 사용하였을 때에는 $T\frac{1}{2}$측 정치에 유의한 차(差)가 없었으며 이 범위 내에서는 Hct., Hb. 및 혈청철치(血淸鐵値)도 역시 유의한 차(差)가 없었다. 3. 1일량(日量) 50ml 및 75ml, 100ml씩 사혈한 군(群)에서는 적혈구(赤血球)만을 사용(使用)하였을 때와 전혈(全血)을 시료(試料)로 하였을 때 사이에 $T\frac{1}{2}$의 측정치에 유의한 차(差)가 있었으며 이 때는 Hct., Hb. 및 혈청철치(血淸鐵値)에도 변화(變化)가 있었다. 즉(卽), 전혈(全血)을 사용한 적혈구(赤血球) 수명($T\frac{1}{2}$)의 측정치가 적혈구(赤血球)만를 사용(使用)한 적혈구(赤血球) 수명($T\frac{1}{2}$)의 측정치 보다 짧았다. 4. 일정(一定)기간(10 일(日)) 사혈의 총량(1000ml)이 같을 매는 200ml를 5회(回) 사혈한 군(群)이나 500ml를 2회(回) 사혈한 군(群) 사이에 적혈구(赤血球) 수명($T\frac{1}{2}$)에 유의(有義)한 차(差)를 볼 수 없었다. 5. 직업적 공혈자의 반복사혈로 인(因)한 만성(慢性) 빈혈환자 9명(名)에서의 $^{51}Cr$적혈구(赤血球)수명($T\frac{1}{2}$) 측정치는 평균(平均) 19.2일(日)로 짧아져 있으나 적혈구수명측정전후(赤血球壽命測定前後)에 충분(充分)한 철제(鐵劑)를 투여(投與)하여 Hct., Hb. 및 혈청철치(血淸鐵値)를 증가(增加)시켰으며 이때 볼 수 있었든 Hct치(値)를 규준(規準)하여 교정한 적혈구(赤血球)수명($T\frac{1}{2}$)은 거의 정상(正常)범위 안에 있어(27.6일(日)) 이러한 인자(因子)를 고려하지 않으면 잘못 이해할 수가 있다. 6. 구충자충(鉤蟲仔蟲)을 구충한 7명(名)의 중등도(中等度) 철(鐵)결핍성 빈혈환자에서의 적혈구(赤血球)수명($T\frac{1}{2}$) 측정치는 25일(日)$\sim$31일(日)로 평균(平均) 28일(日)이었으며, 이때 장 출혈량은 1일(日) $1.0{\sim}3.5ml$이었다. 단시일내의 급성실혈시에는 이와같은 소량의 실혈(失血)도 적혈구(赤血球)수명($T\frac{1}{2}$) 측정치에 영향을 보여 줌을 알 수 있었다. 따라서 이러한 정도의 실혈은 실험오차에 기인하는 것인지 아니면 장기 출혈에서는 이러한 소량의 실혈이 적혈구(赤血球)수명($T\frac{1}{2}$) 측정에 영향을 미치지 않는 것인지는 아직 확실히 말할 수 없다. 8. $^{51}Cr$-표지적혈구(標識赤血球)로 측정한 적혈구(赤血球)수명($T\frac{1}{2}$)은 측정시의 실혈량(失血量)에 큰 영향을 받음을 알 수 있으며 저자(著者)들은 $^{51}Cr$표지적혈구(標識赤血球)를 이용(利用)한 적혈구(赤血球) 수명 측정때 검사기간중 실혈량이 적혈구수명치(赤血球壽命値)에 미치는 관계를 상술(上述)한 실험치(實驗値)를 기초(基礎)로 하여 다음과 같을 교정식(校正式)을 고찰(考察)해 보았다. $^{51}Cr\;T\frac{1}{2}=17.0e^{-0.0495}+18.4e^{-0.000924x}$ 단(但) X : 1일(日) 실혈량(失血量)(단위(單位) ml)

• The Korean Journal of Nuclear Medicine
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• v.33 no.3
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• pp.298-305
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• 1999

Purpose: N-(3-[$^{18}F$]Fluoropropyl)-$2{\beta}$-carbomethoxy-$3{\beta}$-(4-iodophenyl)nortropane [$^{18}F$]FP-CIT) has been shown to be very useful for imaging the dopamine transporter. However, synthesis of this radiotracer is somewhat troublesome. In this study, we used a new method for the preparation of [$^{18}F$]FP-CIT to increase radiochemical yield and effective specific activity. Materials and Methods: [$^{18}F$]FP-CIT was prepared by N-alkylation of nor-${\beta}$-CIT (2 mg) with 3-bromo-1-[$^{18}F$]fluoropropane in the presence of $Et_3N$ (5-6 drops of $DMF/CH_3CN$, $140^{\circ}C$, 20 min). 3-Bromo-1-[$^{18}F$]fluoropropane was synthesized from $5{\mu}L$ of 3-bromo-1-trifluoromethanesulfonyloxypropane (3-bromopropyl-1-triflate) and $nBu_4N^{18}F$ at $80^{\circ}C$. The final compound was purified by reverse phase HPLC and formulated in 13% ethanol in saline. Results: 3-Bromo-1-[$^{18}F$]fluoropropane was obtained from 3-bromopropyl-1-triflate and $nBu_4N^{18}F$ in 77-80% yield. N-Alkylation of nor-${\beta}$-CIT with 3-bromo-1-[$^{18}F$]fluoropropane was carried out at $140^{\circ}C$ using acetonitrile containing a small volume of DMF as the solvents. The overall yield of [$^{18}F$]FP-CIT was 5-10% (decay-corrected) with a radiochemical purity higher than 99% and effective specific activity higher than the one reported in the literature based on their HPLC data. The final [$^{18}F$]FP-CIT solution had the optimal pH (7.0) and it was pyrogen-free. Conclusion: In this study, 3-bromopropyl-1-triflate was used as the precursor for the [$^{18}F$]fluorination reaction and new conditions were developed for purification of [$^{18}F$]FP-CIT by HPLC. We established this new method for the preparation of [$^{18}F$]FP-CIT, which gave high effective specific activity and relatively good yield.

• Nuclear Medicine and Molecular Imaging
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• v.41 no.1
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• pp.9-15
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• 2007

Purpose: Repetitive transcranial magnetic stimulation (rTMS) has recently been clinically applied in the treatment of drug resistant depressed patients. There are mixed findings about the efficacy of rTMS on depression. Furthermore, the influence of rTMS on the physiology of the brain is not clear. We prospectively evaluated changes of regional cerebral blood flow (rCBF) between pre- and post-rTMS treatment in patients with drug resistant depression. Materials and Methods: Twelve patients with drug-resistant depression (7 male, 5 female; age range: $19{\sim}52$ years; mean age: $29.3{\pm}9.3$ years) were given rTMS on right prefrontal lobe with low frequency (1 Hz) and on left prefrontal lobe with high frequency (20 Hz), with 20-minute-duration each day for 3 weeks. Tc-99m ECD brain perfusion SPECT was obtained before and after rTMS treatment. The changes of cerebral perfusion were analyzed using statistical parametric mapping (SPM; t=3.14, uncorrected p<0.01, voxel=100). Results: Following areas showed significant increase in rCBF after 3 weeks rTMS treatment: the cingulate gyrus, fusiform gyrus of right temporal lobe, precuneus, and left lateral globus pallidus. Significant decrement was noted in: the precental and middle frontal gyrus of right frontal lobe, and fusiform gyrus of left occipital lobe. Conclusion: Low-frequency rTMS on the right prefrontal cortex and high-frequency rTMS on the left prefrontal cortex for 3 weeks as an add-on regimen have increased and decreased rCBF in the specific brain regions in drug-resistant depressed patients. Further analyses correlating clinical characteristics and treatment paradigm with functional imaging data may be helpful in clarifying the pathophysiology of drug-resistant depressed patients.

• The Korean Journal of Nuclear Medicine
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• v.37 no.2
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• pp.83-93
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• 2003

Background: Lung-to-heart uptake ratio (LHR) in $^{201}Tl-chloride$ myocardial perfusion scan is believed to be a reliable marker for left ventricular (LV) dysfunction, but the clinical value of LHR is controversial for $^{99m}Tc-MIBI$ imaging. Furthermore, most of results suggesting lung uptake of $^{99m}Tc-MIBI$ as a potential marker for LV dysfunction used immediate post-stress images, instead of routine images acquired 1 hour after tracer injection. The goal of our study was to investigate whether LHR evaluated with routine gated $^{99m}Tc-MIBI$ imaging can reflect the degree of perfusion defect or left ventricular performance. Subjects and Methods: 241 patients underwent exercise $^{99m}Tc-MIBI$ myocardial SPECT were classified into normal myocardial perfusion (NP, n=135) and abnormal myocardial perfusion (AP, n=106) group according to the presence of perfusion defect. LHR was calculated from anterior projection image taken at 1-hour after injection. Two legions of interest (ROIs) were placed on left lung above LV and on myocardium showing the highest radioactivity. Subjects were classified by left ventricular ejection fraction (LVEF), as Gr-I: >50%, Gr-II: 36-50%, Gr-III: <36% and by summed stress score (SSS), as Gr-A: <4, Gr-B: 4-8, Gr-C: 9-13, Gr-D: >13, LHR was compared among these groups. Results: In NP group(n=135), LHR, were higher in men than women ($men:\;0.311{\pm}0.03,\;women:\;0.296{\pm}0.03,\;p<0.05$). Significant difference, in LHR were found between NP and AP groups both for men and women ($men:\;0.311{\pm}0.03\;vs\;.\;0.331{\pm}0.06,\;women:\;0.296{\pm}0.03\;vs.\;0.321{\pm}0.07.\;p<0.05$). There were weak negative correlation between LHR and LVEF (r=-0.342, p<0.05) and weak positive correlation between LHR and SSS (r=0.478, p<0.05) in men, but not in women (LVEF: r=-0.279, p=0.100, SSS: r=0.276, p=0.103). Increased LHR was defined when for more than mean + 2SD value ($men{\geq}0.38,\;women{\geq}0.37$) of the LHR of the subject with normal perfusion. Increased LHR were observed more frequently in subjects with lower LVEF (Gr-I: 11.1%, Gr-II: 27.0%, Gr-III: 35.4%, p<0.05) and higher SSS(Gr-A: 14.0%, Gr-B: 5.7%, Gr-C: 18.2%, Gr-D: 40.7%, p<0.05). Conclusions: LHRs obtained from routine $^{99m}Tc-MIBI$ gated SPECT images were weakly correlated with LVEF and perfusion defect. Although significant overlaps were observed between normal and abnormal perfusion group, LHRs could be used as an indirect marker of severe perfusion defect or reduced left ventricular function.

• Nuclear Medicine and Molecular Imaging
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• v.43 no.1
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• pp.10-18
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• 2009

Purpose: We investigated quantification of dopaminergic transporter (DAT) and serotonergic transporter (SERT) on $^{123}I$-FP-CIT SPECT for differentiating between multiple systemic atrophy (MSA) and idiopathic Parkinson's disease (IPD). Materials and Methods: N-fluoropropyl-$2{\beta}$-carbomethoxy-$3{\beta}$-4-[$^{123}I$]-iodophenylnortropane SPECT ($^{123}I$-FP-CIT SPECT) was performed in 8 patients with MSA (mean age: $64.0{\pm}4.5yrs$, m:f=6:2), 13 with early IPD (mean age: $65.5{\pm}5.3yrs$, m:f=9:4), and 12 healthy controls (mean age: $63.3{\pm}5.7yrs$, m:f=8:4). Standard regions of interests (ROls) of striatum to evaluate DAT, and hypothalamus and midbrain for SERT were drawn on standard template images and applied to each image taken 4 hours after radiotracer injection. Striatal specific binding for DAT and hypothalamic and midbrain specific binding for SERT were calculated using region/reference ratio based on the transient equilibrium method. Group differences were tested using ANOVA with the postHoc analysis. Results: DAT in the whole striatum and striatal subregions were significantly decreased in both patient groups with MSA and early IPD, compared with healthy control (p<0.05 in all). In early IPD, a significant increase in the uptake ratio in anterior and posterior putamen and a trend of increase in caudate to putamen ratio was observed. In MSA, the decrease of DAT was accompanied with no difference in the striatal uptake pattern compared with healthy controls. Regarding the brain regions where $^{123}I$-FP-CIT binding was predominant by SERT, MSA patients showed a decrease in the binding of $^{123}I$-FP-CIT in the pons compared with controls as well as early IPD patients (MSA: $0.22{\pm}0.1$ healthy controls: $0.33{\pm}0.19$, IPD: $0.29{\pm}0.19$), however, it did not reach the statistical significance. Conclusion: In this study, the differential patterns in the reduction of DAT in the striatum and the reduction of pontine $^{123}I$-FP-CIT binding predominant by SERT could be observed in MSA patients on $^{123}I$-FP-CIT SPECT. We suggest that the quantification of SERT as well as DAT using $^{123}I$-FP-CIT SPECT is helpful to differentiate parkinsonian disorders in early stage.

• The Korean Journal of Nuclear Medicine Technology
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• v.14 no.1
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• pp.83-89
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• 2010

Purpose: The Wide Beam Reconstruction (WBR) algorithms that UltraSPECT, Ltd. (U.S) has provides solutions which improved image resolution by eliminating the effect of the line spread function by collimator and suppression of the noise. It controls the resolution and noise level automatically and yields unsurpassed image quality. The aim of this study is WBR of whole body bone scan in usefulness of clinical application. Materials and Methods: The standard line source and single photon emission computed tomography (SPECT) reconstructed spatial resolution measurements were performed on an INFINA (GE, Milwaukee, WI) gamma camera, equipped with low energy high resolution (LEHR) collimators. The total counts of line source measurements with 200 kcps and 300 kcps. The SPECT phantoms analyzed spatial resolution by the changing matrix size. Also a clinical evaluation study was performed with forty three patients, referred for bone scans. First group altered scan speed with 20 and 30 cm/min and dosage of 740 MBq (20 mCi) of $^{99m}Tc$-HDP administered but second group altered dosage of $^{99m}Tc$-HDP with 740 and 1,110 MBq (20 mCi and 30 mCi) in same scan speed. The acquired data was reconstructed using the typical clinical protocol in use and the WBR protocol. The patient's information was removed and a blind reading was done on each reconstruction method. For each reading, a questionnaire was completed in which the reader was asked to evaluate, on a scale of 1-5 point. Results: The result of planar WBR data improved resolution more than 10%. The Full-Width at Half-Maximum (FWHM) of WBR data improved about 16% (Standard: 8.45, WBR: 7.09). SPECT WBR data improved resolution more than about 50% and evaluate FWHM of WBR data (Standard: 3.52, WBR: 1.65). A clinical evaluation study, there was no statistically significant difference between the two method, which includes improvement of the bone to soft tissue ratio and the image resolution (first group p=0.07, second group p=0.458). Conclusion: The WBR method allows to shorten the acquisition time of bone scans while simultaneously providing improved image quality and to reduce the dosage of radiopharmaceuticals reducing radiation dose. Therefore, the WBR method can be applied to a wide range of clinical applications to provide clinical values as well as image quality.

• The Korean Journal of Nuclear Medicine
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• v.38 no.4
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• pp.294-299
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• 2004

Purpose: Both human NIS and mutant $D_2R$ transgenes are proposed as reporting system in transplanted cell tracking. Using hepatoma cell lines, we constructed a dual reporter system containing human sodium-iodide symporter (hNIS) and dopamine 2 receptor ($D_2R$) and compared its characteristics. Materials and Methods: The recombinant plasmid ($pIRES-hNIS/D_2R$) was constructed with IRES (internal ribosome entry site) under control of the CMV promoter $pIRES-hNIS/D_2R$ was transfected to human hepatoma SK-Hep1 cell line with lipofectamine. HEP-ND ($SK-Hep1-hNIS/D_2R$) cells stably expressing hNIS and $D_2R$ was established by selection with G418 for two weeks. RT-PCR was performed to investigate the expression of both hNIS and $D_2R$ genes. The expressions of hNIS and $D_2R$ were measured by $^{125}I$ uptake assays and receptor binding assays. Specific binding of $D_2R$ to $[^3H]spiperone$ was verified by Scatchard plot with (+) butaclamol as a specific inhibitor. $K_d\;and\;B_{max}$ values were estimated. The correlation between hNIS and $D_2R$ expression was compared by using each clone. Results: Similar quantities of hNIS and $D_2R$ genes were expressed on HEP-ND as RT-PCR assays. HEP-ND cells showed 30 to 40 fold higher radioiodine uptakes than those of parental SK-Hep1 cells. $^{125}I$ uptake in HEP-ND cells was completely inhibited by $KClO_4$, a NIS inhibitor Specific binding to HEP-ND cells was saturable and the $K_d\;and\;B_{max}$ values for HEP-ND cells were 2.92 nM, 745.25 fmol/mg protein and 2.91nM, 1323 fmole/mg protein in two clones, respectively. The radioiodine uptake by hNIS activity and $D_2R$ binding was highly correlated. Conclusion: We developed a dual positron and gamma imaging reporter system of hNIS and $D_2R$ in a stably transfected cell line. We expect that $D_2R$ and hNIS genes can complement mutually as a nuclear reporting system or that $D_2R$ can be used as reporter gene when hNIS gene were used as a treatment gene.

• Nuclear Medicine and Molecular Imaging
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• v.42 no.5
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• pp.383-393
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• 2008

Purpose: Cancer specific killing can be achieved by therapeutic gene activated by cancer specific promotor. Expression of sodium iodide symporter (NIS) gene causes transportation and concentration of iodide into the cell, therefore radioiodine treatment after NIS gene transfer to cancer cell could be a form of radionuclide gene therapy. luciferase (Luc) gene transfected cancer cell can be monitored by in vivo optical imaging after D-luciferin injection. Aims of the study are to make vector with both therapeutic NIS gene driven by AFP promoter and reporter Luc gene driven by CMV promoter, to perform hepatocellular carcinoma specific radiodiodine gene therapy by the vector, and assessment of the therapy effect by optical imaging using luciferase expression. Materials and Methods: A Vector with AFP promoter driven NIS gene and CMV promoter driven Luc gene (AFP-NIS-CMV-Luc) was constructed. Liver cancer cell (HepG2, Huh-7) and non liver cancer cell (HCT-15) were transfected with the vector using liposome. Expression of the NIS gene at mRNA level was elucidated by RT-PCR. Radioiodide uptake, perchlorate blockade, and washout tests were performed and bioluminescence also measured by luminometer in these cells. In vitro clonogenic assay with 1-131 was performed. In vivo nuclear imaging was obtained with gamma camera after 1-131 intraperitoneal injection. Results: A Vector with AFP-NIS-CMV-Luc was constructed and successfully transfected into HepG2, Huh-7 and HCT-15 cells. HepG2 and Huh-7 cells with AFP-NIS-CMV-Luc gene showed higher iodide uptake than non transfected cells and the higher iodide uptake was totally blocked by addition of perchlorate. HCT-15 cell did not showed any change of iodide uptake by the gene transfection. Transfected cells had higher light output than control cells. In vitro clonogenic assay, transfected HepG2 and Huh-7 cells showed lower colony count than non transfected HepG2 and Huh-7 cells, but transfected HCT-15 cell did not showed any difference than non transfected HCT-15 cell. Number of Huh-7 cells with AFP-NIS-CMV-Luc gene transfection was positively correlated with radioidine accumulation and luciferase activity. In vivo nuclear imaging with 1-131 was successful in AFP-NIS-CMV-Luc gene transfected Huh-7 cell xenograft on nude mouse. Conclusion: A Vector with AFP promoter driven NIS and CMV promoter driven Luc gene was constructed. Transfection of the vector showed liver cancer cell specific enhancement of 1-131 cytotoxicity by AFP promoter, and the effect of the radioiodine therapy can be successfully assessed by non-invasive luminescence measurement.