微环DNA表达抗EpCAM/抗CD3双特异BiTE分子的抗卵巢癌细胞体外实验研究

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摘要目的探讨抗EpCAM/抗CD3 BsAb分子蛋白对体外卵巢癌细胞生长的抑制效果。方法采用非病毒载体微环DNA表达抗EpCAM/抗CD3 BsAb分子蛋白。观察不同浓度（1×10-3、1×10-4、1×10-5、1×10-6、1×10-7、1×10-8 μg/mL）抗EpCAM/抗CD3 BsAb分子蛋白加至卵巢癌细胞及T细胞中，经6 h和12 h培育后，利用倒置荧光显微镜观察活细胞与死亡细胞数量及形态变化。结果倒置显微镜下可见加抗体后卵巢癌细胞与T细胞明显聚集成块，周围见大量死亡细胞。相同时间下，随着浓度的增加，T细胞介导的卵巢癌肿瘤细胞的杀伤效果并不会一直增强。当浓度达到1×10-4 μg/mL时，培育6 h和12 h时的杀伤均达到最大，差异均有统计学意义（P < 0.05）。结论新型的微环DNA表达系统所表达的抗EpCAM/抗CD3 BsAb对体外卵巢癌细胞生长具有抑制作用，未来可作为一种有效的抗卵巢癌肿瘤的靶向治疗药物。

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	陈远群1 谌平2 陈国创2 韦枝红3

关键词 ： EpCAM, CD3, 双特异BiTE分子蛋白, 卵巢癌, 靶向治疗

Abstract：Objective To study the effect of anti-EpCAM/anti-CD3 BsAb protein inhibition on ovarian cancer cells in vitro. Methods Non-viral vector microcircle DNA was used to obtain the anti-EpCAM/anti-CD3 BsAb protein. Different concentrations (1×10-3, 1×10-4, 1×10-5, 1×10-6, 1×10-7, 1×10-8 μg/mL) of anti-EpCAM/anti-CD3 BsAb protein was added to ovarian cancer cells and T cells. After 6 h and 12 h, the number of live cells and dead cells were counted and morphological changes were observed by inverted fluorescence microscopy. Results Under the inverted microscope, ovarian cancer cells and T cells were obviously aggregated after the addition of anti-EpCAM/anti-CD3 BsAb, and a large number of dead cells were found around them. At the same time, the killing effect of ovarian cancer cells mediated by T cells did not always increased with the increase of concentration. When the concentration reached 1×10-4 μg/mL, the killing rate reached the maximum in both 6 h and 12 h time periods, and the difference were both statistically significant (P < 0.05). Conclusion The anti-EpCAM/anti-CD3 BsAb expressed by the novel microcircle DNA expression system can inhibit the growth of ovarian cancer cells in vitro, and it may be used as an effective targeted anti-ovarian cancer tumor in the future.

Key words： EpCAM CD3 Bispecific BiTE molecular protein Ovarian cancer Targeted therapy

基金资助:国家自然科学基金资助项目（81702581、817005 31）；
广东省深圳市科创委自由探索基础研究基金项目（JC YJ20170306091400043）。

通讯作者: 韦枝红（1966.10-），女，主任医师；研究方向：妇科内分泌学以及卵巢癌靶向治疗。

作者简介: 陈远群（1983.11-），女，硕士；研究方向：盆底学以及卵巢癌靶向治疗。

引用本文:

陈远群1 谌平2 陈国创2 韦枝红3. 微环DNA表达抗EpCAM/抗CD3双特异BiTE分子的抗卵巢癌细胞体外实验研究[J]. 中国医药导报, 2019, 16(30): 12-16.
CHEN Yuanqun1 CHEN Ping2 CHEN Guochuang2 WEI Zhihong3. Experimental study on anti- ovarian carcinoma effect of minicircle DNA expressing anti-EpCAM/anti-CD3 bispecific BiTE in vitro. 中国医药导报, 2019, 16(30): 12-16.

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http://www.yiyaodaobao.com.cn/CN/ 或 http://www.yiyaodaobao.com.cn/CN/Y2019/V16/I30/12

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