West China Hospital/West China School of Nursing, Sichuan University, Chengdu, 610041, China.
Department of Public Health Laboratory Sciences, West China School of Public Health, Sichuan University, Chengdu, 610041, China.
Chin J Integr Med. 2019 Jun;25(6):425-430. doi: 10.1007/s11655-018-2983-5. Epub 2018 May 22.
To examine the effect of the aqueous extract of Ligustrum robustum on tumor growth in vitro and in vivo and explore the possible molecular mechanisms.
In in vitro study, cell viabilities of human cervical carcinoma cells (HeLa), human breast cancer cells (MCF-7), human prostate cancer cells (PC-3), human hepatoma cells (7721) and human colon carcinoma cells (SW480) were evaluated with cell counting kit-8. For L. robustum-treated Hela cells, early or late apoptosis were evaluated by annexin V/PI staining. Mitochondrial membrane potential was measured by staining cells with JC-1. Apoptosis was monitored by nuclear morphology based on chromatin condensation and fragmentation by 4',6-diamidino-2-phenylinole (DAPI) staining. Caspase-3 and -8 activity levels were measured by a colorimetric assay. In vivo, to evaluate the possible mechanism of L. robustum-mediated antitumor effect, nude mouse xenograft study was also conducted.
In in vitro study, L. robustum was found to be toxic to HeLa, MCF-7, PC-3, 7721, SW480, with an half maximal inhibitory concentration value of 2-5 mg/mL (P<0.05). Moreover, externalization of phosphatidylserine, loss of mitochondrial membrane potential, DNA fragmentation and activation of caspase-3 and -8 were detected in L. robustum-treated Hela cells. Using a nude mouse model bearing Hela xenografts, we found that L. robustum reduced tumor volume and tumor weight (P<0.05), but had no effect on body weight and histological damage of important organs. Intraperitoneal injection of L. robustum caused a significant reduction in serum aspartate transaminase and alanine transaminase levels (P<0.05). Furthermore, cleaved caspase-3-positive and terminal nucleotidyl transferase-mediated nick end labeling (TUNEL)-positive cells were observed in L. robustum-treated tumor tissues.
L. robustum inhibits tumor cell growth both in vitro and in vivo by inducing apoptosis in a caspase-dependent way without apparent hepatic toxicity and histological damage, which may offer partial scientific support for the ethnopharmacological claims of L. robustum as a herbal tea for its antitumor activity.
研究女贞子水提物对肿瘤生长的体内外抑制作用，并探讨其可能的分子机制。
在体外研究中，采用细胞计数试剂盒-8 检测人宫颈癌（HeLa）细胞、人乳腺癌（MCF-7）细胞、人前列腺癌（PC-3）细胞、人肝癌（7721）细胞和人结肠癌细胞（SW480）的细胞活力。对于女贞子处理的 HeLa 细胞，通过 Annexin V/PI 染色评估早期或晚期细胞凋亡。通过 JC-1 染色测量线粒体膜电位。通过 4',6-二脒基-2-苯基吲哚（DAPI）染色基于染色质浓缩和碎裂监测细胞凋亡。通过比色法测定半胱天冬酶-3 和 -8 的活性水平。在体内，进行裸鼠异种移植研究以评估女贞子介导的抗肿瘤作用的可能机制。
在体外研究中，发现女贞子对 HeLa、MCF-7、PC-3、7721、SW480 具有毒性，其半数最大抑制浓度值为 2-5mg/mL（P<0.05）。此外，在女贞子处理的 HeLa 细胞中检测到磷脂酰丝氨酸外翻、线粒体膜电位丧失、DNA 片段化以及 caspase-3 和 -8 的激活。使用携带 HeLa 异种移植的裸鼠模型，我们发现女贞子降低了肿瘤体积和肿瘤重量（P<0.05），但对体重和重要器官的组织损伤没有影响。腹腔注射女贞子导致血清天门冬氨酸转氨酶和丙氨酸转氨酶水平显著降低（P<0.05）。此外，在女贞子处理的肿瘤组织中观察到 cleaved caspase-3 阳性和末端核苷酸转移酶介导的 nick 末端标记（TUNEL）阳性细胞。
女贞子通过依赖半胱天冬酶的方式诱导细胞凋亡，从而在体内外抑制肿瘤细胞生长，没有明显的肝毒性和组织损伤，这为女贞子作为一种具有抗肿瘤活性的草药茶的民族药理学说法提供了部分科学依据。
