Downregulation of membrane complement inhibitors CD55 and CD59 by siRNA sensitises uterine serous carcinoma overexpressing Her2/neu to complement and antibody-dependent cell cytotoxicity in vitro: implications for trastuzumab-based immunotherapy.
Downregulation of membrane complement inhibitors CD55 and CD59 by siRNA sensitises uterine serous carcinoma overexpressing Her2/neu to complement and antibody-dependent cell cytotoxicity in vitro: implications for trastuzumab-based immunotherapy.
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DOI：
10.1038/bjc.2012.132
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发表时间：
2012-04-24
影响因子：
8.8
通讯作者：
Santin, A. D.
中科院分区：
文献类型：
--
作者：
Bellone, S.;Roque, D.;Cocco, E.;Gasparrini, S.;Bortolomai, I.;Buza, N.;Abu-Khalaf, M.;Silasi, D-A;Ratner, E.;Azodi, M.;Schwartz, P. E.;Rutherford, T. J.;Pecorelli, S.;Santin, A. D.
我们评估了CD 46、CD 55和CD 59膜结合补体调节蛋白（mCRP）在原发性子宫浆液性癌（USC）中的表达，以及针对这些mCRP的小干扰RNA（siRNA）在体外使USC对补体依赖性细胞毒性（CDC）和抗体（曲妥珠单抗）依赖性细胞毒性（ADCC）敏感的能力。膜结合补体调节蛋白的表达采用实时PCR（RT-PCR）和流式细胞术进行评估，而Her 2/neu表达和c-erbB 2基因扩增采用免疫组织化学，流式细胞术和荧光原位杂交进行评估。在CDC和ADCC 4小时铬释放试验中评价了siRNA介导的mCRP敲低对HER 2/neu过表达USC细胞系的生物学效应。与正常子宫内膜细胞相比，USC细胞中mCRPs的表达明显增高（P<0.05）。RT-PCR和流式细胞仪检测结果显示，抗mCRP siRNA能有效降低USC上CD 46、CD 55和CD 59的表达（P<0.05）。针对USC细胞系的基线补体依赖性细胞毒性（CDC）较低（平均值±s.e.m.= 0.001）。CD 55和CD 59基因敲低后，CD 55和CD 59基因敲低重要的是，在不存在补体的情况下，CD 55和CD 59，而不是CD 46，敲低显著增强了针对过表达Her 2/neu的USC的ADCC。子宫浆液性癌表达高水平的mCRP CD 46、CD 55和CD 59。小干扰RNA抑制CD 55和CD 59（而非CD 46）可使USC在体外对CDC和ADCC敏感，如果特异性靶向肿瘤细胞，则可显著增加曲妥珠单抗介导的体内治疗效果。
We evaluated the expression of CD46, CD55 and CD59 membrane-bound complement-regulatory proteins (mCRPs) in primary uterine serous carcinoma (USC) and the ability of small interfering RNA (siRNA) against these mCRPs to sensitise USC to complement-dependent cytotoxicity (CDC) and antibody (trastuzumab)-dependent cellular cytotoxicity (ADCC) in vitro. Membrane-bound complement-regulatory proteins expression was evaluated using real-time PCR (RT–PCR) and flow cytometry, whereas Her2/neu expression and c-erbB2 gene amplification were assessed using immunohistochemistry, flow cytometry and fluorescent in-situ hybridisation. The biological effect of siRNA-mediated knockdown of mCRPs on HER2/neu-overexpressing USC cell lines was evaluated in CDC and ADCC 4-h chromium-release assays. High expression of mCRPs was found in USC cell lines when compared with normal endometrial cells (P<0.05). RT–PCR and FACS analyses demonstrated that anti-mCRP siRNAs were effective in reducing CD46, CD55 and CD59 expression on USC (P<0.05). Baseline complement-dependent cytotoxicity (CDC) against USC cell lines was low (mean±s.e.m.=6.8±0.9%) but significantly increased upon CD55 and CD59 knockdown (11.6±0.8% and 10.7±0.9%, respectively, P<0.05). Importantly, in the absence of complement, both CD55 and CD59, but not CD46, knockdowns significantly augmented ADCC against USC overexpressing Her2/neu. Uterine serous carcinoma express high levels of the mCRPs CD46, CD55 and CD59. Small interfering RNA inhibition of CD55 and CD59, but not CD46, sensitises USC to both CDC and ADCC in vitro, and if specifically targeted to tumour cells, may significantly increase trastuzumab-mediated therapeutic effect in vivo.
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影响因子：
5.6
作者：
HENDRICKSON, M;ROSS, J;KEMPSON, R
通讯作者：
KEMPSON, R
DOI：
10.1016/j.ijgo.2008.04.008
发表时间：
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作者：
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