Uptake and Cytotoxicity Characterization of Radioiodine in MCF-7 and SKBR3 Breast Cancer Cell Lines

Radioiodine is an effective and low-risk therapy modality in well-differentiated thyroid cancer patients post near-total thyroidectomy. Extra thyroidal tumors such as breast cancer are known to be able to uptake radioiodine. The aim of this study was to analyze the uptake, efflux and cytotoxicity of radioiodine for two molecular types of breast cancer cell lines. Two types of breast cancer cell lines were used in this study, MCF-7 (luminal A type) and SKBR3 (HER2 type). The HaCaT cell line was used as normal cells. Iodine-125 (I-125)was used to measured radioiodine uptake and efflux. Clonogenic assay was used to assess cytotoxicity of iodine-131 (I-131) based on the tested cell reproductive ability. The radioiodine uptake in SKBR3cells was found to be higher than that of MCF-7 and HaCaT cells atp<0.05. The reproductive ability of MCF-7 cells are lower than SKBR3 cells at p<0.05. Both breast cancer cells have lessreproduction ability than HaCaT cells at p<0.05. Both types of breast cancer cells present the ability to uptake radioiodine and show a high sensitivity to radioiodine exposure. Normal cells also demonstrate an ability to uptake radioiodine. However, they have a better tolerance to the amount of I-131 exposure. These findings could potentially lead to the use if I-131 for ablative therapy in breast cancer, similiar to its use in the treatment of thyroid cancer.Received: 4 October 2015; Revised: 14 August 2016; Accepted: 2 September 2016

Synthesis of Sulochrin-125I and Its Binding Affinity as α-Glucosidase Inhibitor using Radioligand Binding Assay (RBA) Method

Most of diabetes patients have type 2 diabetes mellitus or non insulin dependent diabetes mellitus. Treatment of type 2 diabetes mellitus can be conducted by inhibiting the α-glucosidase enzyme which converts carbohydrates into glucose. Sulochrin is one of the potential compounds which can inhibit the function of α-glucosidase enzyme. This study was carried out to obtain data of sulochrin binding with α-glucosidase enzyme as α-glucosidase inhibitor using Radioligand Binding Assay (RBA) method. The primary reagent required in RBA is the labeled radioactive ligand (radioligand). In this study, the radioligand was sulochrin-125I. Prior to the sulochrin-125I synthesis, sulochrin-I was synthesized. Sulochrin-I and sulochrin-125I were synthesized and their bindings were studied using RBA. The  molecular formula of the synthesized Sulochrin-I is C17H15O7I and its molecular weight is 457.9940. The sulochrin-125I was synthesized from sulochrin-I by isotope exchange method. From RBA, the dissociation constant (Kd) and the maximum binding (Bmax) were found as 26.316 nM and 9.302 nM, respectively. The low Kd indicates that sulochrin is capable of binding to α-glucosidase.Received: 08 November 2013; Revised: 28 February 2014; Accepted: 28 March 201

Fragmentation of Nimotuzumab for Preparation of 125I-F(ab’)2-Nimotuzumab as a Precursor for Preparing 125I-F(ab’)2-Nimotuzumab-NLS Radiopharmaceutical for Cancer Therapy

Nimotuzumab is an anticancer agent which belongs to the inhibitor group of Epidermal Growth Factor Receptor (EGFR). Thismonoclonal antibody has a relatively high molecular weight which slowspenetration on tumor cells, making it less attractive in imaging kinetics and potentially elicits antibodies responses. Therefore, in this study nimotuzumab was fragmented to form a bivalent antibody [F(ab’)2] and then labeled with 125I to form 125I-F(ab’)2-nimotuzumab which can be used further as a precursor for preparing 125I-F(ab’)2-nimotuzumab-NLS(NLS = nuclear localization sequence) radiopharmaceuticalfor radioimmunotherapy. The aims of this study was to obtain  characteristics of 125I-F(ab’)2-nimotuzumab by comparing with the 125I labeled-intact nimotuzumab (125I-nimotuzumab). This study was initiated by purifying nimotuzumab by mean of dialysis. The purified nimotuzumab was then fragmented by using pepsin. The F(ab')2-nimotuzumab formed was then purified from its by-products which formed in fragmentation process by using a PD-10 column (consisted Sephadex G25). The intact nimotuzumab and its F(ab’)2 fragment were then labeled with the 125I to form 125I-nimotuzumab and 125I-F(ab’)2-nimotuzumab. The radiochemical purity are 98.27 % and 93.24 %, respectively. Stability test results show that, both 125I-nimotuzumab and 125I-F(ab’)2-nimotuzumab are more stable at 4 °C than at room temperature storage and 37 °C.Received: 24 May 2013; Revised: 21 February 2014; Accepted: 28 February 201

Optimization of Thyroglobulin Coated Tube for Thyroglobulin IRMA Kit

Immunoradiometric assay (IRMA) is a method of analysis based on immunological reactions of antigens-antibodies binding. This highly specific and sensitive method was used for in vitro diagnosis in small quantity of sample. Center for Radioisotope and Radiopharmaceutical Technology, BATAN has developed Thyroglobulin IRMA Kit using coated tube method that can determine thyroglobulin levels in microgram quantities. Coated tube was made with immobilisation of anti thyroglobulin into polistyrene tube. Development of IRMA kit performed through several steps including: optimization component of kit, optimization assay and kit validation. Optimization of coated tube involved selection and volume of solvent, using blocking and non-blocking agent, and volume of blocking agent. The optimum condition for coated tubes was found to be using 0.1M phosphate buffer pH 7.4 with coating volume of 500 μL, 3% BSA in 500 μL blocking agent 0.1M phosphate buffer pH 7.4, with maximum binding and non-specific binding (NSB) of 60.58 and 1.40%, respectively. The optimized coated tube was found to be stable up to 4 weeks

Uptake and Cytotoxicity Characterization of Radioiodine in MCF-7 and SKBR3 Breast Cancer Cell Lines

Radioiodine is an effective and low-risk therapy modality in well-differentiated thyroid cancer patients post near-total thyroidectomy. Extra thyroidal tumors such as breast cancer are known to be able to uptake radioiodine. The aim of this study was to analyze the uptake, efflux and cytotoxicity of radioiodine for two molecular types of breast cancer cell lines. Two types of breast cancer cell lines were used in this study, MCF-7 (luminal A type) and SKBR3 (HER2 type). The HaCaT cell line was used as normal cells. Iodine-125 (I-125)was used to measured radioiodine uptake and efflux. Clonogenic assay was used to assess cytotoxicity of iodine-131 (I-131) based on the tested cell reproductive ability. The radioiodine uptake in SKBR3cells was found to be higher than that of MCF-7 and HaCaT cells atp<0.05. The reproductive ability of MCF-7 cells are lower than SKBR3 cells at p<0.05. Both breast cancer cells have lessreproduction ability than HaCaT cells at p<0.05. Both types of breast cancer cells present the ability to uptake radioiodine and show a high sensitivity to radioiodine exposure. Normal cells also demonstrate an ability to uptake radioiodine. However, they have a better tolerance to the amount of I-131 exposure. These findings could potentially lead to the use if I-131 for ablative therapy in breast cancer, similiar to its use in the treatment of thyroid cancer

Fragmentation of Nimotuzumab for Preparation of 125I-F(ab’)2-Nimotuzumab as a Precursor for Preparing 125I-F(ab’)2-Nimotuzumab-NLS Radiopharmaceutical for Cancer Therapy

Nimotuzumab is an anticancer agent which belongs to the inhibitor group of Epidermal Growth Factor Receptor (EGFR). This monoclonal antibody has a relatively high molecular weight which makes slow penetration on tumor cell, as concequence, it is less attractive in imaging kinetics, and potentially elicits antibodies respons. Therefore in this study nimotuzumab was fragmented to form bivalent antibody [F(ab’)2] and then labeled with 125I to form 125I-F(ab’)2-nimotuzumab which can be used further as a precursor for preparing 125I-F(ab’)2-nimotuzumab-NLS (NLS = nuclear localizing sequences ) radiopharmaceutical for radioimmunotherapy. The aims of this study were to obtain characteristics of 125I-F(ab’)2-nimotuzumab by comparing with the 125I labeled-intact nimotuzumab (125I-nimotuzumab). This study was initiated by purifying nimotuzumab by mean of dialysis. The purified nimotuzumab was then fragmented by using pepsin. The F(ab')2-nimotuzumab formed was then purified from its by-products which formed in fragmentation process by using a PD-10 column (consisted Sephadex G25). The intact nimotuzumab and its F(ab’)2 fragment were then labeled with the 125I to form 125I-nimotuzumab and 125I-F(ab’)2-nimotuzumab. The radiochemical purity are 98.27 % and 93.24 % ,respectively. Stability test results show that, both of 125I-nimotuzumab and 125I-F(ab’)2-nimotuzumab more stable at 4 °C than at room temperature storage and 37 °