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增强自然杀伤细胞对三阴性乳腺癌细胞(MDA-MB-231)的细胞毒性。

augments NK cell cytotoxicity against triple-negative breast cancer cells (MDA-MB-231).

作者信息

Khalaf Taif Kareem, Ismail Norzila, Nazri Nor Amalia, Ahmed Naveed, Yajid Aidy Irman, Mohamud Rohimah, Kadir Ramlah

机构信息

Department of Pharmacology, School of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian, Kelantan, Malaysia.

Department of Medical Microbiology and Parasitology, School of Medical Sciences, Universiti Sains Malaysia, Kubang Kerian, Kelantan, Malaysia.

出版信息

PeerJ. 2024 Nov 28;12:e18420. doi: 10.7717/peerj.18420. eCollection 2024.

Abstract

BACKGROUND

Natural killer cells (NK cells) are essential in cancer immunosurveillance in the body as they can recognize cancer cells that lacking MHC class 1 on their surface. Regulatory cytokines, including interleukin (IL)-18, IL-12, IL-10, IL-8, interferon- (IFN-), and secretory granules like perforin and granzyme are involved in NK cell-mediated cytotoxicity. Stimulating NK cells cytotoxicity towards cancer cells is an ideal strategy to combat cancer naturally. Medicinal plants have been reported to enhance immunity, with () particularly noteworthy due to its abundant bioactive compounds and ability to activate immune cells. This study aimed to evaluate the potential of methanol extract of leaves (MEPB) for enhancing NK cell cytotoxicity against triple-negative human breast cancer cells (MDA-MB-231).

METHODS

The optimal concentration of MEPB to activate NK cells was determined using healthy blood samples, assessing the expression of IL-12, IL-18, IL-10, IL-8, IFN-, perforin, and granzyme B an enzyme-linked immunosorbent assay (ELISA). NK cell purity from healthy donors and breast cancer patients was determined using specific antibodies, and the number of NK cells was assessed using flow cytometry and a hemocytometer. A co-culture experiment, ELISA, and apoptosis assay were used to evaluate NK-mediated cytotoxicity pathways.

RESULTS

ELISA data indicated that MEPB at 7.5 µg/ml significantly increased the expression of IFN-, IL-12, IL-18, perforin, and granzyme B while decreasing IL-8 and IL-10 expression after 20 hrs of incubation. The average NK cell purity was 87.09 ± 0.043%. Breast cancer patients exhibited lower NK cell counts than healthy donors. Co-culture experiments demonstrated that NK cells induced apoptosis in MDA-MB-231 breast cancer cells in the presence of MEPB by increasing perforin, granzyme B, and IFN- expression in both healthy donors and breast cancer patients-experimental groups. enhances NK cell activation, promoting the apoptosis of triple-negative human breast cancer cells (MDA-MB-231), suggesting the potential use of MEPB leaves as an anti-cancer immunostimulant.

摘要

背景

自然杀伤细胞(NK细胞)在机体的癌症免疫监视中至关重要,因为它们能够识别表面缺乏主要组织相容性复合体(MHC)I类分子的癌细胞。调节性细胞因子,包括白细胞介素(IL)-18、IL-12、IL-10、IL-8、干扰素-γ(IFN-γ),以及诸如穿孔素和颗粒酶等分泌颗粒,都参与NK细胞介导的细胞毒性作用。刺激NK细胞对癌细胞的细胞毒性是一种天然对抗癌症的理想策略。据报道,药用植物具有增强免疫力的作用,其中[植物名称]尤其值得关注,因为其含有丰富的生物活性化合物且具有激活免疫细胞的能力。本研究旨在评估[植物名称]叶甲醇提取物(MEPB)增强NK细胞对三阴性人乳腺癌细胞(MDA-MB-231)细胞毒性的潜力。

方法

使用健康血液样本确定激活NK细胞的MEPB最佳浓度,通过酶联免疫吸附测定(ELISA)评估IL-12、IL-18、IL-10、IL-8、IFN-γ、穿孔素和颗粒酶B的表达。使用特异性抗体确定健康供体和乳腺癌患者的NK细胞纯度,并使用流式细胞术和血细胞计数器评估NK细胞数量。采用共培养实验、ELISA和凋亡检测来评估NK介导的细胞毒性途径。

结果

ELISA数据表明,在孵育20小时后,7.5μg/ml的MEPB显著增加了IFN-γ、IL-12、IL-18、穿孔素和颗粒酶B的表达,同时降低了IL-8和IL-10的表达。平均NK细胞纯度为87.09±0.043%。乳腺癌患者的NK细胞计数低于健康供体。共培养实验表明,在MEPB存在的情况下,NK细胞通过增加健康供体和乳腺癌患者实验组中穿孔素、颗粒酶B和IFN-γ的表达,诱导MDA-MB-231乳腺癌细胞凋亡。[植物名称]增强NK细胞活化,促进三阴性人乳腺癌细胞(MDA-MB-231)凋亡,表明MEPB叶具有作为抗癌免疫刺激剂的潜在用途。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4117/11608567/6f76e04ae4d3/peerj-12-18420-g001.jpg

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