Transfection of the Human Sodium/Iodide Symporter(NIS) Gene with Liposomes and the Expression of the NIS Protein in Human Lung A549 Cancer Cells

OBJECTIVE To examine the possibility of human sodium iodide symporter （hNIS） protein expression in lung cancer cells. METHODS Human lung A549 cancer cells were thawed and cultured in vitro. The cells were divided into an experimental group transfected with a recombinant pcDNA3-hNIS plasmid and a control group transfected only with a pcDNA3 plasmid. The recombinant plasmid vector encoding the hNIS gene （pcDNA3-hNIS） was amplified, purified and identified. The hNIS gene was followed by DNA sequencing. A Western blot and an immunohistochemical assay were applied to detect the hNIS protein expression in the transfected human lung A549 cancer cells. RESULTS Restriction enzyme digestion and DNA sequencing results showed the size and direction of the inserted gene in the recombinant pcD- NA3-hNIS plasmid was correct. The Western blot method and immunohistochemical analysis showed a positive NIS protein expression in the experimental group. The NIS protein was detected mainly in the cell membranes showing a positive rate up to 70.6% with no expression of the NIS protein in the control group. There was a significant difference between two groups （P=0.000）. CONCLUSION The hNIS gene was transfected effectively into human lung A549 cancer cells mediated by Lipofectamine 2000, and was expressed with its protein in vitro.

Radioiodine therapy for castration-resistant prostate cancer following prostate-specific membrane antigen promoter-mediated transfer of the human sodium iodide symporter

Radioiodine therapy, the most effective form of systemic radiotherapy available, is currently useful only for thyroid cancer because of the thyroid-specific expression of the human sodium iodide symporter （hNIS）. Here, we explore the efficacy of a novel form of gene therapy using prostate-specific membrane antigen （PSMA） promoter-mediated hNIS gene transfer followed by radioiodine administration for the treatment of castration-resistant prostate cancer （CRPC）. The androgen-dependent C33 LNCaP cell line and the androgen-independent C81 LNCaP cell line were transfected by adenovirus. PSMA promoter-hNIS （Ad.PSMApro-hNIS） or adenovirus.cytomegalovirus-hNIS containing the cytomegalovirus promoter （Ad.CMM-hNIS） or a control virus. The iodide uptake was measured in vitro. The in vivo iodide uptake by C81 cell xenografts in nude mice injected with an adenovirus carrying the hNIS gene linked to PSMA and the corresponding tumor volume fluctuation were assessed. Iodide accumulation was shown in different LNCaP cell lines after Ad.PSMApro-hNIS and Ad.CMV-hNIS infection, but not in different LNCaP cell lines after adenovirus.cytomegalovirus （Ad.CMV） infection. At each time point, higher iodide uptake was shown in the C81 cells infected with Ad.PSMApro-hNIS than in the C33 cells （P 〈 0.05）. An in vivo animal model showed a significant difference in 1311 radioiodine uptake in the tumors infected with Ad.PSMApro-hNIS, Ad.CMV-hNIS and control virus （P 〈 0.05） and a maximum reduction of tumor volume in mice infected with Ad.PSMApro-hNIS. These results show prostate-specific expression of the hNIS gene delivered by the PSMA promoter and effective radioiodine therapy of CRPC by the PSMA promoter-driven hNIS transfection.

Radiofrequency induction on sodium/iodide symporter expression of thyroid cancer

Objective; The aim of this study was to investigate the effects of radiofrequency treatment on sodium/iodide symporter expression of thyroid cancer ceils. Methods： In 29 thyroid cancer patients with low or no expression of soda ／ iodide symporter, the radio frequency combined 1311 therapy was used, the whole-body scintigraphy and serum Ig were detected before and after the radiofrequency treatment. Results： The whole-body scintigraphy showed that 4 cases （4/29） before radiofrequenc_y treatment had positive iodine uptake, 19 cases （19/29） two weeks after radiofrequency treatment had the positive iodine uptake, 12 cases （12/29） four weeks after radiofrequency treatment had the positive iodine uptake. Four weeks after radiofrequency treatment, 5 cases had increased serum Ig levels, 17 cases had decreased serum Ig levels, 7 cases showed no change. 25 cases （25/29） were effective, 15 cases （15/29） were cured. Conclusion： The radiofrequency induced the non-expressed the sodium/iodide symporter of thyroid cancer cells regain the iodine intake ability, it improved the clinical efficacy of 131I therapy in dedifferentiated thyroid cancer.

Correlation analysis between serum β2-MG and sodium/iodide symporter in patients with thyroid carcinoma

Objective： The aim of our study was to investigate the correlation between the expression of sodium/iodide symporter, serum levels of β2-MG and prognosis of thyroid carcinoma patients. Methods： Ninty-five cases with thyroid carcinoma, using enzyme-linked immunosorbent assay with double-antibody sandwich to detect the serum β2-MG levels and immunehistochemistry to detect NIS expression of thyroid cancer tissue. Results： Thirty-seven cases showed positive expression of sodium/iodide symporter （38.9%） and 30 cases showed positive expression of β2-MG （31.57%）. There were significant differences of NIS expression （X2 = 8.207, P = 0.017） and β2-MG expression （X2 = 10.121, P = 0.006） between different pathological types of thyroid carcinoma, but there was no correlation between the positive rate of the two research groups （r = -0.546, P = 0.633）. The significant differences was observed in expression of sodium/iodide symporter （X2 = 9.272, P=0.002） and expression of β2-MG （X2 = 4.441, P = 0.035） between the group with neck lymph node metastasis and the group without neck lymph node metastasis and both positive rate was significantly negatively correlated （r = -1.000, P = 0.000）. The significant differences was observed in expression of sodium/iodide symporter （X2 = 9.272, P = 0.002） and expression of β2-MG （X2 = 3.867, P = 0.043） between the group with distant organ metastasis and the group without distant organ metastasis （X2 = 11.985, P = 0.001） and both positive rate was significantly negatively correlated （r = -1.000, P = 0.000）. Conclusion： There are significantly negatively correlated between neck lymph node metastasis, distant organ metastasis and expression of sodium/iodide symporter and expression of β2-MG. Thyroid cancer lymph node and distant organ metastasis, the tumor tissue NIS expression and serum levels of β2-MG is significantly negatively correlated. The detection of serum β2-MG provides clinical reference value for the effects on radionuclide therapy and prognosis assessment of thyroid carcinoma. Serum β2-MG levels is negatively correlated with prognosis in thyroid cancer patients.

Human sodium/iodide symporter gene induced iodine uptake in human lung adenocarcinoma via baculovirus

To investigate human sodium/iodide symporter (hNIS) induced iodine uptake in human lung adenocarcinoma via baculovirus, a recombinant baculovirus encoding hNIS gene was constructed under the control of CMV promoter (Bac-CMV-hNIS). In vitro, baculovirus infected A549 cells accumulated about 27 times more 125I than that of noninfected cells. The 125I uptake was maximal after 30-min incubation of the cells, and efflux of the radioactivity was rapid, with 50% lost during the first 2 min after 125I-containing medium had been replaced by nonradioactive medium. Competition experiments in the presence of sodium perchlorate revealed a dose-dependent decrease of 125I uptake. Bac-CMV-hNIS infected tumor cells were selectively killed by exposure to 131I, as revealed by clonogenic assays. In nude mice, Bac-CMV-hNIS infected A549 cells accumulated more 131I than that of the control monitored by 1-h scintigraphy after 131I administration. The transduction of hNIS gene through baculovirus is sufficient to induce iodine transporting in A549 cells in vitro and in vivo, outlining the potential of this novel tumor gene imaging approach. But a rapid efflux of radioactivity from the tumor was shown in vivo and the in vivo therapy test showed no sign of effect.

Bibracchial Lariat Ethers(Ⅰ)——The Synthesis and Crystal Structure of the N,N'-bis(2-Methoxy-1-Naphthylmethyl)-4,13-Diaza-18-Crown-6 Sodium Iodide Complex

Diaza Crown ethers are versatile complexing reagents, their complexing abilities are dramatically modified by the electron donating atoms on substituents attached to the nitrogen if they can give a three-dimensional cavity as in cryptates. Hence the synthesis and characterization of bibracchial lariat ethers (BIBLES) have received much attention. In the preparation of new BIBLES, a complex Ⅱ was unexpectedly obtained.

Comparison of Aggressiveness Behavior of Chloride and Iodide Solutions on 304 and 304L Stainless Steel Alloys

The aggressive effect of chloride anion in comparison to iodide anion on the pitting corrosion attack of 304 and 304L stainless steel (SS) alloys was investigated by using the cyclic potentiodynamic polarization test at 0.6 M Sodium Halide salts (NaX) solution and different temperatures. The two alloys 304 and 304L SS suffered from severe pitting corrosion at room temperature up to 50°C in a chloride containing solution with the higher resistance observed for 304L in comparison to the 304 while on pits were detected in iodide solution for both alloys. The pitting potentials of the two alloys in 0.6 M NaCl solution reduced with the increase of the temperature. Examination of the alloys’ surfaces was conducted by using the scanning electron microscopes where it revealed that the occurrence of pitting attack seems like hemispherical or irregular pits with different sizes.

Adenovirus-mediated and tumor-specific transgene expression of the sodium-iodide symporter from the human telomerase reverse transcriptase promoter enhances killing of lung cancer cell line in vitro

Background The sodium-iodide symporter （NIS） protein can mediate the active radioiodine uptake.The human telomerase reverse transcriptase （hTERT） promoter is known to be selectively reactivated in majority of tumors and hence could be used for tumor targeting.We constructed a recombinant adenovirus containing the human sodium iodide symporter （hNIS） gene directed by the hTERT promoter, characterized the ability of infected cells in uptaking iodide, and explored the therapeutic efficacy of 131I in a lung cancer cell line in vitro.Methods The hTERT promoter was amplified by PCR from DNA isolated from log-phase HepG2 cells, subcloned into lineralized FL＊-hNIS/pcDNA3, and then the hTERT-hNIS sequence was subcloned into the shuttle plasmid pAdTrack.The recombinant adenovirus Ad-hTERT-hNIS was constructed by AdEasy system.A positive control adenovirusAd-CMV-hNIS and a negative control adenovirus Ad-CMV were created similarly.A549 cells were transduced with recombinant adenoviruses.125I uptake studies and sodium perchlorate suppression studies were used to confirm hNIS expression and function.Toxic effects of 131I on tumor cells were studied by in vitro clonogenic assay.Results We first successfully constructed an adenovirus mediated transgene expression system of the hNIS under the control of hTERT promoter.When infected with recombinant adenovirus constructs expressing hNIS directed by hTERTand CMV-promoters （Ad-hTERT-hNIS and Ad-CMV-hNIS, respectively）, the lung cancer cell line A549 had increased ability to uptake radioiodide up to 23- and 30- fold compared to the control parental cells, respectively.The radioiodide uptake ability of both the Ad-CMV-hNIS and Ad-hTERT-hNIS transduced cell lines were repressed 11-fold by sodium perchlorate （NaCIO4）.The subsequent in vitro clonogenic assay of the infected A549 cell line was further repressed to 23% （Ad-CMV-hNIS） and 30% （Ad-hTERT-hNIS） of the control group after receiving radioiodide for 7 hours （P 〈0.001）.Conclusion Our preliminary study indicates that an adenovirus mediated transgene expression system of the hNIS under the control of hTERT promoter has the potential to become an effective wide-spectrum yet highly specific anti-cancer strategy.

The effects of thyroid-stimulating hormone,estradiol and prolactin on sodium/iodide symporter mRNA expression in mouse lactating mammary gland cells under different iodine levels

Objective The present study investigated the sodium/iodide symporter mRNA expression in mouse lactating mammary gland cells under different iodine levels and the effects of thyroid-stimulating hormone(TSH),estradiol(E2)and prolactin(PRL)on NIS mRNA expression in mouse lactating mammary gland cells.

电针对紫杉醇诱导大鼠神经病理性疼痛的影响

目的观察电针(EA)对紫杉醇诱导神经病理性疼痛大鼠脊髓背角NKCC1、KCC2表达和小胶质细胞活化的影响及其可能机制。方法将48只雄性SD大鼠随机分为溶媒组(Vehicle组)、紫杉醇组(PTX组)、紫杉醇+电针组(PTX+EA组)、紫杉醇+假电针组(PTX+Sham EA组),每组12只。采用腹腔注射PTX的方法建立PTX诱导的神经病理性疼痛大鼠动物模型,建模完成后,PTX+EA组给予“足三里”、“阳陵泉”电针刺激,连续7 d。于紫杉醇注射前2 d和注射后第1、3、5、7、14、21天进行机械撤足阈值和热缩足潜伏期痛行为学测试。利用免疫荧光染色技术和Western blot技术检测脊髓背角组织中钠钾氯联合转运蛋白1(NKCC1)、钾氯联合转运蛋白2(KCC2)和小胶质细胞标志物离子钙结合衔接分子1(Iba1)表达的变化。结果与Vehicle组比较,PTX组大鼠出现双后足机械和热痛觉过敏,脊髓背角组织中NKCC1表达升高和活化的小胶质细胞数增加。与PTX组比较,PTX+EA组大鼠在第14、21天的机械和热痛觉过敏得到显著改善,脊髓背角组织中NKCC1和Iba1表达量降低,4组间KCC2表达差异无统计学意义。结论电针可有效缓解紫杉醇诱导的神经病理性疼痛,其机制可能与抑制大鼠脊髓背角组织中NKCC1表达和小胶质细胞活化有关。

甲状腺癌碘-131治疗抵抗发生的分子机制研究进展

甲状腺癌是内分泌系统最常见的恶性肿瘤,其中分化型甲状腺癌(differentiated thyroid carcinoma,DTC)占90%以上。多数DTC患者经过系统治疗后预后良好,但少数患者肿瘤原发灶或转移灶出现失分化现象,进展为放射性碘难治性DTC(radioiodine-refractory DTC,RAIR-DTC),预后明显变差,是甲状腺癌致死的主要原因。钠碘转运体(sodium iodide symporter,NIS)的表达和功能异常,是导致甲状腺癌碘-131治疗抵抗的主要原因,其发生受遗传学改变、表观遗传学改变、肿瘤微环境作用、自噬作用等多因素影响。遗传学改变如BRAF基因的V600E位点突变、RET/PTC基因重排等导致致癌信号通路的激活,直接或间接地影响NIS的表达及其在细胞膜上的正常定位。表观遗传学调控特定基因的表达模式,调节NIS的表达水平,进而影响甲状腺细胞的碘摄取功能。肿瘤微环境中的免疫细胞、细胞因子和细胞外基质等成分也可能通过降低NIS的表达水平和/或干扰其在细胞膜上的正常功能导致细胞碘摄取障碍。此外,自噬作为一种细胞内部的代谢调节机制,也可以调节NIS的表达及其在细胞内的分布,从而影响碘的摄取和碘-131治疗的敏感性。通过综述以上因素在甲状腺癌失分化中的作用机制,可以更全面地理解RAIR-DTC的发生和发展过程,有助于探寻新的治疗靶点,改善预后,并为患者提供更有效的个体化治疗策略。

钠碘同向转运体表达与功能调控的研究进展

钠碘同向转运体(sodium iodine symporter)广泛分布于人体各个器官,是器官转运I-的结构和功能基础。钠碘同向转运体主要表达在细胞的基底侧膜上,为各种生命活动提供所需的I-。目前已知,钠碘同向转运体的调控机制涉及激素、细胞因子、转录因子及信号分子的作用。此外,钠碘同向转运体作为甲状腺癌的重要治疗靶点逐渐受到关注,然而在治疗过程中唾液腺等其他器官吸收放射性碘而产生的并发症也值得重视。本文重点综述了钠碘同向转运体的表达、作用及其调控机制的研究进展,旨在为临床干预钠碘同向转运体异常相关疾病进程提供新思路。

Triphenylphosphine and sodium iodide: a new catalyst combination to rival precious metal complexes in visible light photoredox catalysis

Photoredox catalysis has changed the landscape of modern synthetic organic chemistry by prompting a dramatic increase in research activity into reactions that proceed through free radical intermediates.

利用示波器测量μ子寿命

μ子是地球表面的主要宇宙射线,可以通过碘化钠等常见闪烁体探测器进行探测.本文介绍了一种利用高采样率的示波器对探测器所捕捉到的μ子信号及其衰变信号进行数字化采集、线下软件分析的实验方法,进而得到μ子寿命.本实验操作相对简单,可以在不使用放射源和传统核电子学的情况下,拓展为近代物理或专业物理实验中的一个粒子探测前沿项目.

基于ZYNQ的γ能谱采集系统研制

本文设计了一种基于ZYNQ的γ能谱测量系统,用于瞬发中子活化分析仪中NaI探测器的γ能谱测量。γ能谱采集系统主要包括高压电源、数字多道幅度分析系统以及上位机等,可实现高达130 Msps的采集频率以及千兆以太网的数据传输及控制。经^(137)Cs、^(60)Co、^(152)Eu等放射源的测试,γ能谱采集系统能谱响应线性拟合优度为0.999,能量分辨率为8.76%(@0.662 keV)。结果表明,该γ能谱采集系统性能达到煤质成分检测的能谱测量要求。

离子色谱法测定9V、11A、19A型肺炎球菌多糖中碘化钠残留量

建立检测9V、11A、19A型肺炎球菌多糖中碘化钠残留量的离子色谱方法,并进行验证。以Dionex IonPac AS11-HC阴离子交换层析柱为分析柱,Dionex IonPac AG11-HC阴离子交换柱为保护柱;采用低浓度氢氧化钠溶液为淋洗液等度洗脱,以电化学检测器进行检测。碘化钠的质量浓度在0.01~1.00μg/mL的范围具有良好的线性关系,决定系数R 2可达到0.9999,检出限为0.002μg/mL,定量限为0.010μg/mL。3个型别肺炎球菌多糖在低、中、高3个加标水平下的平均回收率为94.40%~103.91%,相对标准偏差(RSD)为0.17%~1.71%,均满足检测要求。方法无需复杂的样品前处理过程,测定快速,灵敏度高,结果准确,重复性好,可用于9V、11A、19A型肺炎球菌多糖中痕量碘化钠残留的检测。

Capsaicin restores sodium iodine symportermediated radioiodine uptake through bypassing canonical TSH–TSHR pathway in anaplastic thyroid carcinoma cells

Anaplastic thyroid cancer(ATC)is a rare but highly lethal disease.ATCs are resistant to standard therapies and are extremely difficult to manage.The stepwise cell dedifferentiation results in the impairment of the iodine-metabolizing machinery and the infeasibility of radioiodine treatment in ATC.Hence,reinducing iodine-metabolizing gene expression to restore radioiodine avidity is considered as a promising strategy to fight against ATC.In the present study,capsaicin(CAP),a natural potent transient receptor potential vanilloid type 1(TRPV1)agonist,was discovered to reinduce ATC cell differentiation and to increase the expression of thyroid transcription factors(TTFs including TTF-1,TTF-2,and PAX8)and iodine-metabolizing proteins,including thyroidstimulating hormone receptor(TSHR),thyroid peroxidase,and sodium iodine symporter(NIS),in two ATC cell lines,8505C and FRO.Strikingly,CAP treatment promoted NIS glycosylation and its membrane trafficking,resulting in a significant enhancement of radioiodine uptake of ATC cells in vitro.Mechanistically,CAP-activated TRPV1 channel and subsequently triggered Ca2þinflux,cyclic adenosine monophosphate(cAMP)generation,and cAMP-responsive element-binding protein(CREB)signal activation.Next,CREB recognized and bound to the promoter of SLC5A5 to facilitate its transcription.Moreover,the TRPV1 antagonist CPZ,the calcium chelator BAPTA,and the PKA inhibitor H-89 effectively alleviated the redifferentiation exerted by CAP,demonstrating that CAP might improve radioiodine avidity through the activation of the TRPV1–Ca2þ/cAMP/PKA/CREB signaling pathway.In addition,our study indicated that CAP might trigger a novel cascade to redifferentiate ATC cells and provide unprecedented opportunities for radioiodine therapy in ATC,bypassing canonical TSH–TSHR pathway.

碘对大鼠体内及体外甲状腺钠碘转运体mRNA表达的调节

目的探讨碘过量对体内、体外甲状腺钠碘转运体(NIS)mRNA表达的影响。方法Wistar大鼠,随机分为低碘组(LI)、适碘组(NI)、5倍高碘组(5HI)、10倍高碘组(10HI)、50倍高碘组(50HI)、100倍高碘组(100HI),检测尿碘、甲状腺NISmRNA表达水平。FRTL细胞分别在含有10-6～10-3mol/L碘化钾的培养基中培养24、48h,检测NISmRNA水平的变化。结果LI组尿碘显著低于NI组,而甲状腺NISmRNA表达水平明显高于NI组(P<0.01);各高碘组尿碘与NI组比较呈成倍升高,NISmRNA水平与NI组相比逐渐下降。FRTL细胞在分别含有10-6～10-3mol/L碘化钾的培养基中培养24、48h,NISmRNA的表达水平与对照组差异无统计学意义(P>0.05)。结论碘对体内、体外甲状腺NISmRNA表达的影响存在不同的机制,长期、慢性处于高碘摄入的大鼠主要通过转录水平影响甲状腺NISmRNA的表达,而体外急性实验表明,高碘则可能通过转录后水平而起作用。

维甲酸诱导甲状腺癌细胞摄碘的实验研究

目的 探讨全反式维甲酸(ATRA)诱导甲状腺癌细胞系的钠碘同向转运体(NIS)表达及其碘摄取。方法 通过ATRA诱导甲状腺癌细胞系滤泡状甲状腺癌细胞株(FTC 133)、乳头状甲状腺癌细胞株(W3)及未分化甲状腺癌细胞株( 85 0 5C)后,经逆转录 聚合酶链反应(RT PCR)及Westernblot检测甲状腺癌细胞系的NISmRNA及其蛋白质表达,并测定甲状腺癌细胞系诱导后的摄碘变化。结果 ATRA诱导甲状腺癌细胞系4 8h后,FTC 133和W3的NISmRNA及蛋白质表达增高,85 0 5C未见变化;ATRA诱导甲状腺癌细胞系2周后,FTC 133和W3的摄碘增高。结论 ATRA能诱导分化型甲状腺癌细胞摄碘增高。

甲状腺癌NIS和TSHR表达的矛盾性及非相关性

目的研究钠/碘转运蛋白(sodium/iodide symporter,NIS)和促甲状腺激素受体(thyroid stim-ulating hormone receptor,TSHR)在甲状腺癌中的表达和相关性。方法收集正常甲状腺组织18例、甲状腺癌23例,通过荧光定量分析和免疫组织化学SP法对NIS和TSHR的表达进行研究。结果甲状腺癌NIS和TSHR mRNA的表达显著低于正常甲状腺组织(P<0.005),正常甲状腺组织NIS和TSHR的表达呈显著正相关(r=0.667,P=0.000),而在甲状腺癌中无相关性(r=0.222,P=0.376)。免疫组织化学结果表明甲状腺癌NIS阳性产物主要定位于细胞质,TSHR阳性产物定位于细胞质和细胞膜,甲状腺癌NIS及TSHR蛋白阳性表达率显著高于正常组(P<0.05);甲状腺癌NIS mRNA表达下降同时伴有细胞质NIS蛋白表达升高的有17例(74%)。结论甲状腺癌NIS和TSHR表达具有非相关性以及在转录和翻译水平上具有矛盾性,NIS蛋白非功能定位于细胞质,因而不能发挥正常的运输及聚碘功能,我们推测这可能是造成甲状腺癌患者放射性碘治疗失败的重要原因。