^188Re—IGF-1类似物对胰腺癌细胞Patu8988的增殖抑制与诱导凋亡实验

Effect of ^188 Re-IGF-1 analogue in proliferation inhibition and apoptosis induction in pancreatic carcinoma cells

目的研究^188Re—IGF-1类似物（IGF-1A）对胰腺癌Patu8988细胞的增殖抑制效应和诱导凋亡作用。方法（1）制备^188Re—IGF-1A，用四甲基偶氮唑蓝（MqT）实验检测给药后细胞增殖情况，细胞按给药情况分为对照组、IGF-1A组（1、5、10、20μg）、^188Re04组（0．37、1．85、3．70、7．40MBq）和^188Re—IGF-1A组（0．37、0．74、1．85MBq）。^188Re04组和^188Re—IGF-1A组分别在给药后1～7d，IGF-1A组分别在给药后1～6d，每天用MT＇I＂比色法测定其吸光度（A）值，计算细胞生存率和抑制率。（2）将1×10。个细胞接种于培养瓶中，分^188Re04组和^188Re—IGF-1A组（均设1．85、3．70、7．40MBq3个剂量），在给药后3d用流式细胞仪检测细胞凋亡率。（3）36只荷人胰腺癌裸鼠，瘤内注射^188Re—IGF-12．28—4．81MBq，分别在15min，1、4h，1、3和5d显像后处死裸鼠6只，计算各组织的％ID／g和每MBq活度的吸收剂量（mGy／MBq）。对数据行单因素方差分析。结果（1）IGF-1A和^188Re—IGF-1A能抑制Patu8988细胞生长。加入^188Re—IGF-1A后4d，1．85MBq亚组抑制率达到（90．75±5．20）％，高于相同化学剂量的IGF-1A（5txg）组或相同放射性活度的^188Re04组[（49．50±2．39）％与（23．00±4．21）％]，差异有统计学意义（F=554．724，P〈0．01）。（2）给药1．85、3．70、7．40MBq3d后，^188Re-IGF-1A组漂浮细胞比例分别为（16．56±0．95）％、（33．39±5．93）％和（43．76±1．38）％，漂浮细胞的凋亡率分别为（12．70±2．27）％、（17．80±1．51）％和（23．23±1．22）％。（3）荷瘤鼠瘤内注射^188Re—IGF-1A后15min和1、3、5d肿瘤内放射性摄取分别为（39．30±17．98）、（10．59±9．39）、（5．32±1．53）和（5．30±2．28）％ID／g。肿瘤内吸收剂量为5165．8mGy／MBq。结论^188Re—IGF-1A对胰腺癌细胞生长有明显的抑制作用，并可诱导细胞凋亡；瘤内注射后肿瘤部位摄取较高。

Objective To investigate the effect of ^188Re-IGF-1 analogue （IGF-1A） in proliferation inhibition and apoptosis induction in human pancreatic carcinoma cell line Patu8988. Methods IGF-1A was labeled with ^188Re. Patu8988 cells were divided into an un-treated control group, IGF-1A group （1,5, 10, 20 lag） ,＇SSReO4 group （0.37, 1.85, 3.70, 7.40 MBq） and iSSRe-IGF-1A group （0.37, 0.74, 1.85 MBq）. The cell proliferation inhibition effects by the ^188Re-IGF-1A and ^188ReQ- were detected every day by 3-（4,5- dimethyhhiazol-2-yl）-2,5-diphenyhetrazolium bromide （MTF） test from 1 d to 7 d after administration, while the IGF-1A group was tested every day from 1 d to 6 d after treatment. Inhibition rates were calculated. At 3 d after treatment with lSSReO4 and ISSRe-IGF-1A （1.85, 3.70, 7.40 MBq）, cell apoptosis was detected by flow cytometry. For biodistribution studies of lSS Re-IGF-1 A, 36 nude mice beating Patu8988 cell xenografts were divided into 6 groups. At different time points （ 15 min, 1 h, 4 h, 1 d, 3 d and 5 d）,36 mice （ n = 6 per time point） were sacrificed and organs of interest were removed, weighted and measured for radioactivity by a gamma counter. The absorbed doses of organs were calculated as % ID/g. One-way analysis of variance was used. Results After 4 d, inhibition rate of Patu8988 cell proliferation in the ^188 Re- IGF-1A group （1.85 MBq） was （90.75 ±5.20） %, higher than that in lSSReO4- group or IGF-1A group （ （49.50 ± 2.39）%, （23.00 ± 4.21 ）% ; F = 554.724, P 〈 O. O1 ）. At 3 d after treatment with different doses of ^188Re-IGF-1A（ 1.85, 3.70, 7. dO MBq）, floating cell ratios were （ 16.56 ± 0.95） %, （33.39 ± 5, 93） % and （43.76 ± 1.38） %, respectively. Apoptosis ratios in the floating cells treated by lSSRe-IGF- 1A （1.85, 3.70, 7.40 MBq） were （12.70 ±2.27）%, （17.80 ± 1.51）% and （23.23 ± 1.22）%, respectively. Distribution in tumors was （39.30 ± 17.98）, （ 10.59 ± 9.39）, （5.32 ± 1.53） and （5.30 ± 2.28） % ID/g at the 15 min, 1 d, 3 d, and 5 d timepoints after intratumoral injection, respectively. The absorbed dose of tumors was 5165.8 mGy/MBq. Conclusions Proliferation of human pancreatic carcinoma cell line Patu8988 can be inhibited and apoptosis can also be induced by ^188Re-IGF-1A. The tumor region is the major distribution site in nude mice bearing human pancreatic cancer xenografts after intratumoral injection of ISSRe-IGF-1A.