Jiangsu Key Laboratory for Pharmacology and Safety Evaluation of Chinese Materia Medica, School of Pharmacy, Nanjing University of Chinese Medicine, 210023 Nanjing, Jiangsu, China.
School of Medicine & Holistic Integrative Medicine, Nanjing University of Chinese Medicine, 210023 Nanjing, Jiangsu, China.
Front Biosci (Landmark Ed). 2021 Nov 30;26(11):1106-1118. doi: 10.52586/5013.
: Natural killer (NK) cells play an indispensable role in anti-tumor immunity. TGF-β1 is the main accomplice of tumor immune escape, inhibiting tumor immunity mediated by NK cells. It is reported that Salvia miltiorrhiza can promote the immune killing effect of NK cells. In this study, Tanshinol, a water-soluble active component of Salvia miltiorrhiza, was used to investigate its effect on the inhibition of NK cell functions mediated by TGF-β1 in breast cancer. : We constructed a mouse model of breast cancer by tail vein injection, H&E staining and ELISA were used to verify the role of TGF-β1 and the effects of Tanshinol on breast cancer and NK cells. , we used CCK8 and cytotoxicity assays to preliminarily evaluate the effect of Tanshinol on the anti-tumor effect of NK cells intervention by TGF-β1. We explored the killing activity of NK cells and related signal pathways by immunofluorescence imaging technology, RT-PCR, ELISA and flow cytometry. Also, Western blot, RT-PCR and immunofluorescence experiments were applied to investigate the expression level of the natural killer group 2 member D (NKG2D)-NKG2D ligands (NKG2DL) signal axis, and combined with immunoprecipitation, to detect the formation of NKG2D-DNAX-activating protein of 10 kD (DAP10) complex. : TGF-β1 played a role in promoting lung metastasis of breast cancer and inhibiting the secretion of cytotoxic mediators from NK cells, but Tanshinol could reverse it. High-dose Tanshinol also significantly optimized the survival rate of tumor-bearing mice. TGF-β1 could destroy the NKG2D-NKG2DL axis, down-regulate the expression and nuclear accumulation of p-smad2/3. Moreover, TGF-β1 inhibited the activation of PI3K-ERK1/2-PLCγ2 signaling pathway that is related to the degranulation of NK cells, and diminished the expression of degranulation marker CD107a and the release of anti-tumor cytotoxic killing medium of NK cells. However, Tanshinol was able to interfere with the negative regulation of TGF-β1 on the functions of NK cells, mainly through promoting the expression of NKG2D and its molecular chaperone DAP10, thereby propelling the formation of NKG2D-DAP10 complex. : Collectively, Tanshinol enables NK cells to activate and release multiple killing mediators to carry out immune attacks on tumor cells.
: 自然杀伤 (NK) 细胞在抗肿瘤免疫中发挥着不可或缺的作用。TGF-β1 是肿瘤免疫逃逸的主要帮凶，抑制 NK 细胞介导的肿瘤免疫。有报道称，丹参可以促进 NK 细胞的免疫杀伤作用。在这项研究中，丹参水溶性活性成分丹参醇被用于研究其对 TGF-β1 抑制 NK 细胞功能的影响在乳腺癌中的作用。 : 我们通过尾静脉注射构建了乳腺癌小鼠模型，通过 H&E 染色和 ELISA 验证了 TGF-β1 以及丹参醇对乳腺癌和 NK 细胞的作用。此外，我们使用 CCK8 和细胞毒性测定初步评估了丹参醇对 TGF-β1 干预 NK 细胞抗肿瘤作用的影响。我们通过免疫荧光成像技术、RT-PCR、ELISA 和流式细胞术探讨了 NK 细胞的杀伤活性及相关信号通路。此外，还应用 Western blot、RT-PCR 和免疫荧光实验研究了自然杀伤组 2 成员 D (NKG2D)-NKG2D 配体 (NKG2DL) 信号轴的表达水平，并结合免疫沉淀检测 NKG2D-DNAX-激活蛋白 10kD (DAP10) 复合物的形成。 : TGF-β1 促进乳腺癌肺转移并抑制 NK 细胞细胞毒性介质的分泌，但丹参醇可逆转此作用。高剂量丹参醇还显著提高了荷瘤小鼠的存活率。TGF-β1 可破坏 NKG2D-NKG2DL 轴，下调 p-smad2/3 的表达和核积累。此外，TGF-β1 抑制与 NK 细胞脱颗粒相关的 PI3K-ERK1/2-PLCγ2 信号通路的激活，并降低脱颗粒标记物 CD107a 的表达和 NK 细胞抗肿瘤细胞毒性杀伤介质的释放。然而，丹参醇能够干扰 TGF-β1 对 NK 细胞功能的负调控，主要是通过促进 NKG2D 及其分子伴侣 DAP10 的表达，从而促进 NKG2D-DAP10 复合物的形成。 : 综上所述，丹参醇使 NK 细胞激活并释放多种杀伤介质，对肿瘤细胞进行免疫攻击。
