Purpose: Dual reporter gene imaging has several advantages for more sophisticated molecular imaging studies such as gene therapy monitoring. Herein, we have constructed hepatoma cell line expressing dual reporter genes of sodium iodide symporter (NIS) and enhanced green fluorescence protein (EGFP), and the functionalities of the genes were evaluated in vivo by nuclear and optical imaging. Materials and Methods: A pRetro-PN vector was constructed after separating NIS gene from pcDNA-NIS. RSV-EGFP-WPRE fragment separated from pLNRGW was cloned into pRetro-PN vector. The final vector expressing dual reporter genes was named pRetro-PNRGW. A human hepatoma (HepG2) cells were transfected by the retrovirus containing NIS and EGFP gene (HepG2-NE). Expression of NIS gene was confirmed by RT-PCR, radioiodine uptake and efflux studies. Expression of EGFP was confirmed by RT-PCR and fluorescence microscope. The HepG2 and HepG2-NE cells were implanted in shoulder and hindlimb of nude mice, then fluorescence image, gamma camera image and I-124 microPET image were undertaken. Results: The HepG2-NE cell was successfully constructed. RT-PCR showed NIS and EGFP mRNA expression. About 50% of cells showed fluorescence. The iodine uptake of NIS-expressed cells was about 9 times higher than control. In efflux study, $T_{1/2}$ of HepG2-NE cells was 9 min. HepG2-NE xenograft showed high signal-to-background fluorescent spots and higher iodine-uptake compared to those of HepG2 xenograft. Conclusion: A hepatoma cell line expressing NIS and EGFP dual reporter genes was successfully constructed and could be used as a potential either by therapeutic gene or imaging reporter gene.
Purpose: Multidrug resistance (MDR) of the cancer cells related to mdr1 gene expression can be effectively treated by selective short hairpin RNA for mdr1 gene (shMDR). Sodium/iodide symporter (NIS) gene is well known to have both reporter and therapeutic gene characteristics. We have co-transfected both shMDR and NIS gene into colon cancer cells (HCT15 cell) expressing MDR and Tc-99m sestamibi and I-125 uptake were measured. In addition, cytotoxic effects of doxorubicin and I-131 therapy were also assessed after transfection. Material and Methods: At first, shMDR was transfected with liposome reagent into human embryonic kidney cells (HEK293) and HCT cells. shMDR transfection was confirmed by RT-PCR and western blot analysis. Adenovirus expressing NIS (Ad-NIS) gene and shMDR (Ad-shMDR) were co-transfected with Ad-NIS into HCT15 cells. Forty-eight hours after infection, inhibition of P-gycoprotein (Pgp) function by shMDR was analyzed by a change of Tc-99m sestamibi uptake and doxorubicin cytotoxicity, and functional activity of induced NIS gene expression was assessed with I-125 uptake assay. Results: In HEK293 cells transfected with shMDR, mdr1 mRNA and Pgp protein expressions were down regulated. HCT15 cells infected with 20 MOI of Ad-NIS was higher NIS protein expression than control cells. After transfection of 300 MOI of Ad-shMDR either with or without 10 MOI of Ad-NIS, uptake of Tc-99m sestamibi increased up to 1.5-fold than control cells. HCT15 cells infected with 10 MOI of Ad-NIS showed approximately 25-fold higher I-125 uptake than control cells. Cotransfection of Ad-shMDR and Ad-NIS resulted in enhanced cytotoxic by doxorubicin in HCT15 cells. I-131 treatment on HCT15 cells infected with 20 MOI of Ad-NIS revealed increased cytotoxic effect. Conclusion: Suppression of mdr1 gene expression, retention of Tc-99m sestamibi, enhanced doxorubicin cytotoxicity and increases in I-125 uptake were achieved in MDR expressing cancer cell by co-transfection of shMDR and NIS gene. Dual therapy with doxorubicin and radioiodine after cotransfection shMDR and NIS gene can be used to overcome MDR.
Purpose : The aim of this study was to predict radiation dose at 1 meter with BMI(body mass index) in thyroid cancer patients treated with radio-iodine and provide the efficient guideline in the management of patients. Methods : 140 patients from thyroidectomy for thyroid cancer were enrolled. All subjects under went 150 mCi radio-iodine therapy and performed whole body scan 1 week later. BMI(weight divided by square of height) was calculated to evaluate the amount of fatty tissue indirectly. The radiation dose at 1 meter was measured initially and on 2nd days. the relation of values with BMI were analyzed statically. As for the method of statistical analysis, using Med calc Version 9,2,2,0 Program. Results : (1) The initial effective dose was inversely correlated with the BMI. Significance level was 0.0004. (2) We obtained the following formula from the data of initial effective dose and BMI: Y = -30.91X + 350.4(${\mu}Sv/h$)(Y: initial radiation dose, x: Group). (3) After 21.55 hours, than radiation dose was less than those recommended by ICRP or NRC in 53% of the population. Conclusion : Using BMI, the initial radiation dose and 2nd days dose can be predicted in thyroid cancer patients before radio-iodine therapy. It may be used for predicting the time of discharge and control the isolation room. We were able to predict the radiation exposure after discharge using this calculated value.
Purpose: The green tea polyphenol (GTPP) has been known to exert antioxidant activity as a radical scavenger as well as cancer preventive and cancer growth inhibition effect. The aim of this study was to identify whether GTPP not only potentiate the growth inhibition effect in ${\gamma}-irradiated$ human cancer cell but also exert protection action for irradiated human normal cell. Materials and Methods: GTPP (80% catechin including >45% EGCG) added in the HL60, human leukemia, and NC37, human lymphoblast, before irradiation. After establishing the amount of GTPP and the dose of radiation, the cells were treated with the GTPP for 6 hours and irradiated with the determined doses. Results: Viability when $10{\mu}g/ml$ GTPP added before ${\gamma}-irradiation$ with 1 Gy to NC37 cells was not different in comparison with control but it when was irradiated with 3 Gy significantly different (1 Gy;P=0.126, 3 Gy;P=0.010). $20{\mu}g/ml$ GTPP did not show significant difference in both NC37 cells irradiated with 1 Gy and 3 Gy (1 Gy;P=0.946, 3 Gy;P=0.096). Viabilities were significantly decreased with concentration of additional GTPP in HL60 with 1 or 3 Gy (1 Gy $69.0{\pm}1.7%\;vs\;42.4{\pm}1.3%,\;3\;Gy;\;66.9{\pm}3.9%\;vs\;44.2{\pm}1.6%$). Conclusion: In vitro study, we certified that when the cells were irradiated with dose below 3 Gy, GTPP provide not only anticancerous effect against cancer cells but also radioprotective effect in normal cells simultaneously. Theses results suggest the possibility that consumption of green tea could give the radioprotective effect and maximize the effect on internal radiation such as radioiodine therapy concomitantly.
The 20-minute $^{99m}Tc-pertechnetate$ uptake became readily available for routine use and it replaced $^{131}I$ for thyroid imaging. However measuring thyroid uptake during a 5-minute minimizes pertechnetate uptake by the salivary glands and presence of contaminated saliva from those glands in to the pharynx and esophagus. A study was carried out to determine the suitability of the utility of a S-minute and 20-minute interval from administration of $^{99m}Tc-pertechnetate$ to imaging and uptake measurement as a replacement for the 24 hour standard originally established with $^{131}I$, and to evaluate the relationship between 5-minute $^{99m}Tc-pertechnetate$ uptake and other thyroid functions. A 5-minute and 20-minute uptake of $^{99m}Tc-pertechnetate$ were measured in 70 patients with thyroid disease at Yeungnam University Hospital from March 1, 1991 to Feb. 29, 1992. The results were as follows. 1) The 5-minute $^{99m}Tc-pertechnetate$ uptake in Graves' disease, Hashimoto's thyroiditis, simple goiter, non toxic nodular goiter, subacute thyroiditis and euthyroid were 18.2%, 14.6%, 2.8%, 3.2%, 1.2% and 1.1%, respectively. There was a significant difference between the mean of the euthyroid group and the mean of the Graves' disease. So differenciation between them can be easily made. 2) The 5 minute $^{99m}Tc-pertechnetate$ thyroid uptake was well correlated with 24 hour $^{131}I$ thyroid uptake (r=0.75, p<0.001). These data provided an equation for estimating the 24 hour uptake of iodide given the 5 minute pertechnetate uptake: Estimated 24-hour $^{131}I$ thyroid Uptake= 7.188*ln (5 minute $^{99m}Tc-pertechnetate$ uptake)+16.94 3) The 20-minute $^{99m}Tc-pertechnetate$ thyroid uptake was well correlated with 24-hour $^{131}I$ uptake (r=0.72, p<0.001) and 5-minute $^{99m}Tc-pertechnetate$ thyroid uptake (r=0.96, p<0.001). 4) In the Graves' disease, The 5-minute $^{99m}Tc-pertechnetate$ thyroid uptake was well correlated with serum $T_3-resin$ uptake (r=0.46, p<0.01), serum total $T_3$ (r=0.55, p<0.05), serum total $T_4$ (r=0.46, p<0.05). These results suggest that 5-minute ${99m}Tc-pertechnetate$ thyroid uptake has been found at least as useful as 24-hour $^{131}I$ uptake for diagnostic confirmation at our hospital, the logistical advantages of completing the diagnosis. The exam in 5-minutes led us to abandon the 24-hour study in the majority of patients, but the 24-hour $^{131}I$ uptake is still obtained in patients with planned or potential radioiodine therapy.
Purpose: In general, We discharged radioactive wastewater and sewages less than $8.1{\times}10^{-13}$ Ci/ml in a exclusive water-purifier tank. Our hospital operating three exclusive water-purifier tank for radioactive wastewater and sewages of 60 tons capacity respectively. In order to meet the criteria it need a enough decay more than 125 days per each exclusive tank. However, recently we fell into the serious situation that decay period was decreased remarkably, owing to the wastewater amount increased rapidly by enlarge the therapy ward. For that reason, in this article, I'd like to say the way that reducing of radioactive wastewater and sewages rationally. Materials and Methods: From January, 2006 to October, four hundred and two cases were analyzed. They were all hospitalized during 3 days and 2 nights. We calculated the average amount of water used (include toilet water used, shower water used, washstand water used, $\cdots$), each exclusive water-purifier tank's decay period, as well as try to search the increased factors about water-purifier tank inflow flux by re-analysis of the procedure of radioisotope therapy step by step. Results: We could increase each exclusive water-purifier tank's decay period from 84 days to 130 days through the improvement about following cause: (1) Improvement of conventional toilet stool for excessive water waste $\rightarrow$ Replacement of water saving style toilet stool (2) Prevention of unnecessary shower and wash (3) Stop the diuretics taking during hospitalization (4) Analysis of relationship between water intakes and residual dose of body (5) Education about outside toilet utilization before the administration (6) Changed each water-purifier tank's maximum level from85% to 90% Conclusion: The originality of our efforts are not only software but hardware performance improvements. Incidentally the side of software's are change of therapy procedures and protocols, the side of hardware's are replacement of water saving style toilet stool and change of each water-purifier tank's maximum level. Thus even if a long lapse of time, problem such as return to the former conditions may not happen. Besides, We expect that our trials become a new reasonable model in similar situation.
To evaluate the utility of I-131, T1-201 scintigraphy, and serum thyroglobulin(Tg) in the follow-up of differentiated thyroid cancer, we compared retrospectively the data from 33 patients who underwent total or subtotal thyroidectomy. I-131 scintigraphy was performed after optimal endogenous TSH stimulation ($TSH>50{\mu}U/ml$). Total 41 cases of I-131 and T1-201 scintigraphy pairs were examined. Concomitant serum thyroglobulin levels were measured for 41 pairs of scan. Tg-off levels(that measured after discontinuation of the thyroid hormone) higher than 40ng/m1 were considered positive, and Tg-on levels(that measured during the thyroid hormone replacement) higher than 5ng/ml were considered positive. The concordance rates between I-131 therapeutic scintigraphy and T1-201 scintigraphy was 48% in the 38 case of total scan pairs(59% in the 17 cases of postoperative preablation group, and 38% in the 21 cases of postoperative postablation group). Of 17 studies before the I-131 ablation therapy(preablation group), 7 showed positive I-131 therapeutic scintigraphy despite of negative T1-201 scintigraphy. Among patients with negative I-131 therapeutic scintigraphy, no patients had abnormal T1-201 uptake. However, of 21 studies which were done after radioiodine therapy(postablation group) 6 had abnormal uptake on T1-201 scintigraphy which were not seen on I-131 therapeutic scintigraphy, and Tg-off levels also elevated in this 4 of 6 cases. As a result, I-131 therapeutic scintigraphy showed highest positive rate at postoperative preablation follow-up study in differentiated thyroid cancer patients. T1-201 scintigraphy may be useful in postablation studies, and the use of the combined modalities(T1-201 and Tg levels) provides a higher diagnostic yield.
Purpose: Idoxifene is currently entering phase II clinical trials for the treatment of advanced breast cancer. The radiolabeled idoxifene using $[^{123}I]$ provides an opportunity for clinical pharmacology with single photon emission computed tomography (SPECT). The purpose of this study was to prepare radiolabeled idoxifene using $[^{123}I]$ and to determine its cell uptake of breast cancer cell line. Materials and Methods: With a view to evaluating new anticancer drugs, we are investigating the novel antiestrogen pyrrolidino-4-iodotamoxifen (idoxifene). $[^{123}I]$Idoxifene has been prepared in no-carrier-added form using a tributyl stannylated precursor which has been synthesized by means of (2-chloroethoxy)benzene with (${\pm}$)-2-phenylbutanoic acid on the basis of previously reported standard methods. The biodistribution and dynamic behavior of the compound were investigated using the comparative breast cancer cell line, MCF-7 (estrogen receptor-positive) and MDA-MB-468 (non-estrogen receptor). Results and Conclusion: Acylation of (2-chloroethoxy)benzene with (${\pm}$)-2-phenylbutanoic acid gave the versatile ketone (81%) which reacted with 1,4-diiodobenzene to give triphenylethylene as a mixture of E and Z geometric isomers, which were separated by the recrystallization in ethanol. The E-isomer was treated with pyrrolidine to give idoxifene (67%). In order to incorporate radioactive iodine into the 4-position, the 4-stannylated precursor was prepared (30%). The yield of radioiodination was 90-92% with a high radiochemical purity greater than 98%. The ratio of tumor uptake of the breast cancer cell line between MCF-7 and MDA-MB-468 was about 1.7.
Purpose: Human Na+/I- symporter (hNIS) is known to be expressed in many tissues other than thyroid gland. The breast cancer cells are one of them and the possibility of radioiodine therapy in treatment of the breast cancer may be suggested. We investigated the expression rate of hNIS and the relationship between the expression of hNIS and the finding of 99mTc-MIBI scintimammographv in the breast cancer Materials and Methods: Surgically proved 56 patients with breast cancer were the subjects of this study The expression of hNIS were evaluated by immunohistochemistry and the results were compared to the findings of 99m7c-MIBI scintimammography. Results: Overall expression rate of hNIS was 41.1% in 56 patients. According to the pathologic diagnosis, it was 42.9% in 49 patients with invasive ductal carcinoma and 28.6% in the 7 patients with ductal carcinoma in situ. The expression rate of hNIS in the 41 cases with a focal increased uptake at he breast lesion on 99m7c-MIBI sointimammogram was 31.7%. That in the 15 cases without any abnormal uptake on the scan was significantly higher(65.7%, p<0.05). Conclusion: The expression rate of hNIS in the patients with breast cancer was not so high. The rate was higher in the patients with no increased uptake at the breast lesion on 99m7c-MIBI scintimammography.
Purpose: A prospective comparison was made between imaging with Tc-99m pertechnetate (Tc-99m) and Ioine-131 (I-131) for the detection of residual and metastatic tissue after total thyroidectomy in patients with well-differentiated thyroid carcinoma. Materials and Methods: Initially our patients had imaging with Tc-99m, followed by I-131 within 3 days. The study included 21 patients who had ablation with high dose of I-131 ranging from 100 mCi to 150 mCi. Planar and pinhole images were acquired for both Tc-99m and I-131. Diagnostic images of Tc-99m and I-131 were compared with post-therapy images. Degree of uptake on Tc-99m and I-131 images was scored by four point scale and compared. Results: The results of the Tc-99m study were: 16 of 19 studies (84%) were positive on simple planar images, but 19 of 20 studies (95%) were positive on pinhole images. Conventional I-131 diagnostic imaging on the other hand showed that all studies (100%) were positive on both planar and pinhole images. There was a significant difference in degree of uptake between Tc-99m and I-131 planar images (p<0.05). Only one case of Tc-99m scintigraphy was negative on both planar and pinhole studies (false negative). There was no distant metastasis on the therapeutic I-131 images. Conclusion: Tc-99m scan using pinhole in certain clinical situations is an alternative to the I-131 scan in detecting thyroid or lymph node metastasis prior to the first ablative treatment after thyroidectomy for well-differentiated thyroid carcinoma.
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