The In vivo99mTc-HYNIC-annexinV

apoptosis imaging has been reported to be able to predict the severity of myocardium infarction, organ transplantation rejection and response to tumor chemotherapy treatment [5, 6]. Encouraging results were reported by some pilot studies [7, 8] that early phase99mTc-HYNIC-annexin V scintigraphy (TAVS) after radiotherapy in patients may be useful as a predictive test for treatment response. However, the potential value of99mTc-HYNIC-annexin V imaging in the evaluation of radiation-induced buy ACP-196 apoptosis has yet to be established. In order to evaluate the value of99mTc-HYNIC-annexin V imaging in detecting early phase apoptosis in tumors after single dose irradiation and in predicting tumor response 4SC-202 solubility dmso to radiotherapy, a radiation murine tumor model was NVP-LDE225 supplier established,

and the relevance of TAVS image to apoptosis and radiation sensitivity was explored. Methods Animals Male C57BL/6 mice and Kunming mice were obtained from the breeding facility of the Experimental Animal Center, West China Medical Center, Sichuan University. All mice were used between 6 and 12 weeks of age, and weighed 18 to 22 g. Care of all experimental animals was in accordance with institutional guidelines and approved protocols. Cell Culture Technique The C57BL/6 mice derived EL4 lymphoma cell line was obtained from the Transplantation Immunology Laboratory of West China Hospital, Sichuan University. The Kunming mice derived S180 sarcoma cell line was obtained from the Tumor Biotherapy Laboratory of West China Hospital, Sichuan University. Both EL4 and S180 cell lines were grown as cell suspensions in RPMI 1640 medium, supplemented with 10% (v/v) fetal bovine serum and 290 μg/mL L-glutamine, 100 U/mL penicillin and 100 μg/mL streptomycin.

Cells were maintained in the logarithmic growth phase at a concentration of 1-5 × 105 cells/mL at 37°C in a 5% CO2 in air Acyl CoA dehydrogenase atmosphere under aseptic conditions. Flow cytometry (FCM) assessment of apoptosis Groups of EL4 lymphoma cells in logarithmic growth phase were irradiated with a single dose of: 0 Gy, 2 Gy, 4 Gy or 8 Gy; the S180 sarcoma cells received only 0 Gy or 8 Gy. The 0 Gy group was served as the unirradiated control for both tumors. Irradiation was with 4 MV X-rays generated by the Elekta Precise linear accelerator (Elekta, Sweden) using 100 cm SSD,10 cm × 10 cm portal size, with the cell culture flask lying on a 1.0 cm thick Perspex. Twenty-four hours after irradiation, the samples were harvested and stained with Annexin V-FITC and PI for 15 min at 25°C by using a commercial kit (BD Pharmingen, USA). Cells were washed twice with PBS and re-suspended in buffer solution (1 × 106 cells per ml). Stained cells were analyzed with a flow cytometer (BD, FACSAria™) within 1 hour of staining, as described in the manufacturer’s manual.