Pham Dang-Huan, Kim Ju-Sun, Kim Sang-Ki, Shin Dong-Jun, Uong Nguyen-Thanh-Tung, Hyun Hoon, Yoon Mee Sun, Kang Sin Jae, Ryu Young Jae, Cho Jin Seong, Yoon Jung Han, Lee Ji Shin, Cho Duck, Lee Soo-Hyeon, Park Min Ho
Department of Biomedical Sciences, Chonnam National University Medical School, Gwangju, Republic of Korea.
Department of Companion & Laboratory Animal Science, Kongju National University, Yesan, Republic of Korea.
Anticancer Res. 2017 Oct;37(10):5507-5513. doi: 10.21873/anticanres.11981.
BACKGROUND/AIM: The inhibition of a disintegrin and metalloproteinase (ADAM) has the potential to become a novel approach for natural killer (NK) cell-based cancer immunotherapy. Thus, the aim of this study was to investigate the influence of ADAM10 and ADAM17 inhibitors on expanded NK cell to enhance antibody-dependent cellular cytotoxicity (ADCC) in breast cancer cell lines.
NK cells were expanded in medium supplemented with an ADAM10 or ADAM17 inhibitor to prevent the shedding of soluble CD16/FcγRIII. The expression level of CD16 and production of interferon-gamma (IFN-γ) was detected by flow cytometry using specific antibodies. ADCC activity of expanded NK cells was estimated in trastuzumab treated breast cancer cell lines such as MCF-7, MDA-MB-231, SKBR3, and BT-474 cells.
The ADAM17 inhibitor increased the purity of expanded NK cells to 90% after 14 days at 5 and 10 μM in vitro (p=0.043). However, the expansion rate of NK cells was decreased at 10 μM of the ADAM 17 inhibitor (p=0.043). Inhibition of ADAM10 suppressed the expansion of NK cells, although the NK purity was increased at 1 μM of the inhibitor. The expression of CD16 was significantly increased at 1 and 5 μM of the ADAM17 inhibitor (p=0.046, 0.028, respectively) during the culturing period. Inhibition of ADAM10 reduced the expression of CD16 on NK cells. The cytotoxic activity of the ADAM17 inhibitor treated NK cells against MCF-7 (p=0.039) and BT-474 (p=0.027) cells was significantly elevated. The ADCC activity of NK cells treated with 5 μM of ADAM17 inhibitor was significantly increased against SKBR-3 and BT-474 (p=0.027). Inhibition of ADAM17 increased the production of IFN-γ in expanded NK cells.
The inhibition of ADAM17 enhanced the purity of expanded NK cells and the ADCC activity of these cells against trastuzumab treated breast cancer cell lines.
背景/目的：抑制解聚素和金属蛋白酶（ADAM）有可能成为基于自然杀伤（NK）细胞的癌症免疫治疗的新方法。因此，本研究的目的是探讨ADAM10和ADAM17抑制剂对扩增的NK细胞的影响，以增强其对乳腺癌细胞系的抗体依赖性细胞毒性（ADCC）。
在添加了ADAM10或ADAM17抑制剂的培养基中扩增NK细胞，以防止可溶性CD16/FcγRIII脱落。使用特异性抗体通过流式细胞术检测CD16的表达水平和干扰素-γ（IFN-γ）的产生。在曲妥珠单抗处理的乳腺癌细胞系（如MCF-7、MDA-MB-231、SKBR3和BT-474细胞）中评估扩增的NK细胞的ADCC活性。
在体外，14天后，5 μM和10 μM的ADAM17抑制剂可使扩增的NK细胞纯度提高到90%（p=0.043）。然而，10 μM的ADAM 17抑制剂会降低NK细胞的扩增率（p=0.043）。抑制ADAM10会抑制NK细胞的扩增，尽管在1 μM抑制剂时NK细胞纯度有所提高。在培养期间，1 μM和5 μM的ADAM17抑制剂可使CD16的表达显著增加（分别为p=0.046、0.028）。抑制ADAM10会降低NK细胞上CD16的表达。ADAM17抑制剂处理的NK细胞对MCF-7（p=0.039）和BT-474（p=0.027）细胞的细胞毒性活性显著提高。用5 μM的ADAM17抑制剂处理的NK细胞对SKBR-3和BT-474的ADCC活性显著增加（p=0.027）。抑制ADAM17可增加扩增的NK细胞中IFN-γ的产生。
抑制ADAM17可提高扩增的NK细胞的纯度以及这些细胞对曲妥珠单抗处理的乳腺癌细胞系的ADCC活性。
