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In vitro and in vivo anti-cancer activities of Kuding tea (Ilex kudingcha C.J. Tseng) against oral cancer

机译:苦丁茶(Ilex kudingcha C.J. Tseng)对口腔癌的体外和体内抗癌活性

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Kuding tea (Ilex kudingcha C.J. Tseng) is drunk widely in China. The in vitro anticancer effects of Kuding tea were evaluated in TCA8113 human tongue carcinoma cells using a 3-(4,5-dimethyl-2-thiazolyl)-2,5- diphenyltetrazolium bromide (MTT) assay. At a concentration of 200 μg/ml, Kuding tea exhibited an inhibitory effect of 75% in TCA8113 cells, which was higher than that observed at concentrations of 100 and 50 μg/ml (41 and 10% inhibition, respectively). Reverse transcription-polymerase chain reaction (RT-PCR) and western blot analyses of the apoptosis, inflammation and metastasis genes and proteins in Kuding tea-treated cancer cells were performed. Kuding tea significantly induced apoptosis in TCA8113 cancer cells (P<0.05) by upregulating Bax, caspase-3 and caspase-9 expression, and downregulating Bcl-2 expression. Expression of the NF-κB, iNOS and COX-2 genes that are associated with inflammation was significantly downregulated by Kuding tea, which demonstrated its anti-inflammatory properties. Kuding tea also exerted an anti-metastatic effect on cancer cells. This was demonstrated by the decreased expression of matrix metalloproteases (MMPs) and the increased expression of tissue inhibitors of metalloproteinases (TIMPs), and confirmed by the inhibition of the metastasis of U14 squamous cell carcinoma cells in imprinting control region (ICR) mice. The ICR mouse buccal mucosa cancer model was established by injecting the mice with U14 cells. Following injection, the wound at the injection site was topically treated with Kuding tea. It was observed that the tumor volumes for the group treated with Kuding tea were smaller than those from the control mice. Analysis of the sections of buccal mucosa cancer tissue demonstrated that the buccal mucosa cancer degrees of the Kuding tea-treated mice were weaker than that in the control mice. Similar results were observed in the lesion sections of the cervical lymph nodes. Based on these results, Kuding tea exhibited successful in vitro anticancer effects in TCA8113 cells and in vivo buccal mucosa cancer preventive activity.
机译:苦丁茶(Ilex kudingcha C.J. Tseng)在中国广泛饮用。使用3-(4,5-二甲基-2-噻唑基)-2,5-二苯基四唑溴化物(MTT)分析评估了TCA8113人舌癌细胞中苦丁茶的体外抗癌作用。在200μg/ ml的浓度下,苦丁茶对TCA8113细胞的抑制作用为75%,高于在100和50μg/ ml的浓度下(分别为41%和10%)的抑制作用。进行逆转录-聚合酶链反应(RT-PCR)和蛋白质印迹分析,以分析苦丁茶处理过的癌细胞的凋亡,炎症和转移基因和蛋白质。苦丁茶通过上调Bax,caspase-3和caspase-9的表达以及下调Bcl-2的表达,显着诱导TCA8113癌细胞凋亡(P <0.05)。苦丁茶显着下调了与炎症有关的NF-κB,iNOS和COX-2基因的表达,证明了其具有抗炎作用。苦丁茶还对癌细胞具有抗转移作用。基质金属蛋白酶(MMPs)的表达减少和金属蛋白酶组织(TIMPs)的表达增加证明了这一点,并通过抑制印迹控制区(ICR)小鼠中的U14鳞状细胞癌细胞的转移得到了证实。通过向小鼠注射U14细胞来建立ICR小鼠颊粘膜癌模型。注射后,用苦丁茶局部治疗注射部位的伤口。观察到用苦丁茶治疗的组的肿瘤体积小于对照小鼠的肿瘤体积。对颊粘膜癌组织切片的分析表明,苦丁茶治疗小鼠的颊粘膜癌程度较对照小鼠弱。在子宫颈淋巴结的病变部分观察到类似的结果。基于这些结果,苦丁茶在TCA8113细胞中显示出成功的体外抗癌作用,并且在体内具有颊粘膜癌的预防活性。

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