【摘要】 目的:探究放射性¹²⁵I粒子联合Pictilisib（GDC-0941）对卵巢透明细胞癌的杀伤及抗肿瘤免疫效应的影响。方法:将人卵巢透明细胞癌细胞株ES-2分为对照组、¹²⁵I粒子组(细胞接受¹²⁵I粒子照射处理)、GDC-0941组（1μmol/L GDC-0941培养细胞24 h）及¹²⁵I+GDC-0941组(细胞接受¹²⁵I粒子照射处理，并以1μmol/L GDC-0941培养24 h),按照分组进行处理后，MTT法测定细胞存活率，流式细胞术检测细胞凋亡率，Hoechst 33258染色观察细胞凋亡形态；皮下接种ES-2建立卵巢透明细胞癌裸鼠移植瘤模型，并随机分为对照组、¹²⁵I粒子组(将¹²⁵I粒子植入肿瘤组织中心)、GDC-0941组（灌胃125 mg/kg的GDC-0941）及¹²⁵I+GDC-0941组(将¹²⁵I粒子植入肿瘤组织中心并灌胃125 mg/kg的GDC-0941),给药开始后，每隔2 d测量并计算肿瘤体积，21 d后处死裸鼠并取其肿瘤组织，电子天平称重，HE染色观察肿瘤组织内细胞生长情况，免疫组织化学染色检测肿瘤组织内凋亡蛋白Cleaved-caspase-3与增殖标记物Ki-67的表达，流式细胞术检测外周血T淋巴细胞亚群。结果:相较于¹²⁵I粒子照射和GDC-0941单独处理，¹²⁵I与GDC-0941联合作用下，细胞存活率下降（P<0.05）,细胞凋亡率增加（P<0.05）,且细胞核浓缩明显，浓染颗粒较多，碎裂现象严重。与¹²⁵I粒子植入和GDC-0941灌胃的两组裸鼠比较，¹²⁵I粒子植入联合GDC-0941灌胃的裸鼠，肿瘤生长较为缓慢，肿瘤块变小、质量也减小（P<0.05）,肿瘤组织内细胞排列稀疏，Cleaved-caspase-3阳性表达率升高（P<0.05）,Ki-67阳性表达率降低（P<0.05）,此外，外周血中CD3⁺CD8⁺T细胞比例增加。结论:放射性¹²⁵I粒子联合GDC-0941作用能够显著提高对卵巢透明细胞癌的杀伤作用，且抑制肿瘤生长并改善移植瘤裸鼠的抗肿瘤免疫功能，两者联合作用效果要优于单独作用。更多还原

【Abstract】 Objective:To explore the effect of radioactive ¹²⁵I particles combined with Pictilisib（GDC-0941） on the killing and anti-tumor immune effects of ovarian clear cell carcinoma.Methods:The cultured human ovarian clear cell cancer cell line ES-2 was randomly divided into control group, ¹²⁵I particle group(cells received ¹²⁵I particle irradiation treatment),GDC-0941 group（1 μmol/L GDC-0941 cultured cells for 24 h） and ¹²⁵I+GDC-0941 group(cells received ¹²⁵I particle irradiation treatment and cultured at 1 μmol/L GDC-0941 for 24 h),after processing according to the grouping, MTT method was used to measure cell survival rate, flow cytometry was used to measure cell apoptosis rate, Hoechst 33258 staining was used to observe cell apoptosis morphology.A nude mouse xenograft model of ovarian clear cell carcinoma was established by subcutaneous inoculation of ES-2.The 16 nude mice that were successfully modeled were randomly divided into a control group, a ¹²⁵I particle group(¹²⁵I particles were implanted into the tumor tissue center),and a GDC-0941 group（GDC-0941 with 125 mg/kg gavage） and ¹²⁵I+GDC-0941 group(GDC-0941 with ¹²⁵I seeds implanted into the center of tumor tissue and gavage 125 mg/kg GDC-0941),after the administration started, the tumor volume was measured and calculated every 2 days.After 21 days, the nude mice were sacrificed and their tumor tissues were taken, weighed with an electronic balance, HE staining was used to observe the cell growth in the tumor tissues, immunohistochemical staining was used to detect the expression of apoptosis protein Cleaved-caspase-3 and proliferation marker Ki-67 in tumor tissues, flow cytometry was used to detectperipheral blood T lymphocyte subsets.Results:Compared with ¹²⁵I particle irradiation and GDC-0941 treatment alone, the combined effect of ¹²⁵I and GDC-0941 decreased the cell viability（P<0.05） and increased the apoptosis rate（P<0.05）.The nuclei were condensed obviously, the densely stained granules were more, and the fragmentation phenomenon was serious.Compared with the two groups of nude mice implanted with ¹²⁵I seeds and gavaged with GDC-0941,the nude mice implanted with ¹²⁵I seeds and gavaged with GDC-0941 had slower tumor growth, smaller tumor mass and reduced mass（P<0.05）,the cells in the tumor tissue were sparsely arranged, the positive expression rate of Cleaved-caspase-3 was increased（P<0.05）,and the positive expression rate of Ki-67 was decreased（P<0.05）.In addition, the proportion of CD3⁺CD8⁺T cells in peripheral blood increased（P<0.05）.Conclusion:The combination of radioactive ¹²⁵I particles and GDC-0941 can significantly increase the killing effect on ovarian clear cell carcinoma, inhibit tumor growth and improve the anti-tumor immune function of transplanted tumor nude mice.The combined effect of the two is better than the single effect.更多还原