Department of Biological Treatment Center, 105th Hospital of PLA, Hefei 230031, Anhui, China.
Central Laboratory of the First Affiliated Hospital of Anhui Medical University, Hefei 230035, Anhui, China.
Cancer Biomark. 2017 Dec 6;20(4):609-615. doi: 10.3233/CBM-170588.
To investigate the improvement of cytotoxicity of autologous CIKs from patients with breast cancer to MCF-7 cells by suppressed PD-1 expression.
The Lentiviral Vector/PD-1 carrying the gene that can suppressed PD-1 was transferred to CIK cells from patients with breast cancer to inhibit PD-1 expression. The PD-1 protein were detected by RT-PCR and Western blot. The positive PD-1 of CIKs and PD-L1 of MCF-7 cells were detected by FCM, and cytotoxicity of CIKs to MCF-7 was assayed by CCK-8.
The PD-1 positive CIKs with Lentiviral Vector/PD-1 transferred from patients with breast cancer were 16.02%, 14.36% and 14.64% at 14th, 21st and 28th day, obviously inhibited as compared to 50.54%, 74.50% and 73.36% in CIKs without transinfection (P< 0.05); the Lentiviral Vector/PD-1 decreased the PD-1 mRNA levels in CIK cells, and Lentiviral Vector/PD-1-transferred CIKs had lower PD-1 expression; CCK-8 detection showed that at 14th day, the cytotoxicity rates of CIKs with blank plasmids and those with PD-1 transfection to MCF-7 cells were 58.78% and 68.14%, respectively.
MCF-7 cells have a strong PD-L1 expression at its surface, and inhibition of PD-1 expression can improve the cytotoxicity of CIK cells.
通过抑制 PD-1 表达，研究提高乳腺癌患者自体 CIK 对 MCF-7 细胞的细胞毒性。
将携带能抑制 PD-1 基因的慢病毒载体/PD-1 转染入乳腺癌患者的 CIK 细胞，抑制 PD-1 表达。通过 RT-PCR 和 Western blot 检测 PD-1 蛋白。通过 FCM 检测 CIK 细胞的 PD-1 阳性和 MCF-7 细胞的 PD-L1 阳性，通过 CCK-8 测定 CIK 对 MCF-7 的细胞毒性。
从乳腺癌患者转染慢病毒载体/PD-1 的 PD-1 阳性 CIKs 在第 14、21 和 28 天分别为 16.02%、14.36%和 14.64%，明显低于未转染的 CIKs（50.54%、74.50%和 73.36%）（P<0.05）；慢病毒载体/PD-1 降低了 CIK 细胞中的 PD-1 mRNA 水平，并且转染了慢病毒载体/PD-1 的 CIKs 的 PD-1 表达较低；CCK-8 检测显示，在第 14 天，空白质粒转染的 CIK 和 PD-1 转染的 CIK 对 MCF-7 细胞的细胞毒性率分别为 58.78%和 68.14%。
MCF-7 细胞表面具有很强的 PD-L1 表达，抑制 PD-1 表达可以提高 CIK 细胞的细胞毒性。
