目的 探究香菇多糖联合顺铂通过调控I-κB激酶β（IKKβ）/核因子-κB（NF-κB）通路对乳腺癌的作用机制。方法 采用完全培养基将香菇多糖胶囊制备成10、20、40 μg/mL的溶液，将顺铂制备成10 μmol/L溶液。选用含有10%胎牛血清的RPMI-1640培养液进行细胞培养，将其分为模型组、顺铂组（10 μmol/L），顺铂＋香菇多糖组在顺铂组处理的基础上，分别加10、20、40 μg/mL香菇多糖，培养48 h用于体外细胞实验，CCK-8法检测乳腺癌MCF-7细胞增殖情况，计算细胞增殖抑制率；流式细胞术检测细胞凋亡情况。随机将大鼠分为模型组、顺铂组（2 mg/kg）、香菇多糖组（15 mg/kg）、顺铂（2 mg/kg）＋香菇多糖（15 mg/kg）组、Prostratin（0.5 mg/kg）组，每组10只。将培养的乳腺癌细胞依次注射于各组大鼠乳腺垫内进行造模。各组大鼠进行相应的给药处理，每日ip 1次，模型组大鼠ip等剂量的生理盐水，连续干预28 d。计算各组大鼠瘤体体积、数量并称重；Western blotting法检测各组大鼠NF-κB p65、NF-κB p-p65、IKKβ表达水平。结果 与模型组相比，顺铂组、顺铂＋香菇多糖（10、20、40 μg/mL）组乳腺癌细胞增殖抑制率、细胞凋亡率显著升高（P<0.05）；与顺铂组相比，顺铂＋香菇多糖（10、20、40 μg/mL）乳腺癌细胞增殖抑制率、细胞凋亡率显著升高（P<0.05），且呈剂量相关性增长。与模型组相比，顺铂组、香菇多糖组、顺铂＋香菇多糖组大鼠瘤体体积、瘤体质量、瘤体数目、瘤体内NF-κB p-p65/NF-κB p65、IKKβ蛋白表达量显著降低（P<0.05）；与顺铂组、香菇多糖组相比，顺铂＋香菇多糖组大鼠瘤体体积、瘤体质量、瘤体数目、瘤体内NF-κB p-p65/NF-κB p65、IKKβ蛋白表达量显著降低（P<0.05）；与顺铂＋香菇多糖组相比，Prostratin组大鼠瘤体体积、瘤体质量、瘤体数目、瘤体内NF-κB p-p65/NF-κB p65、IKKβ蛋白表达量显著升高（P<0.05）。结论 顺铂联合香菇多糖可通过调控IKKβ/NF-κB通路，抑制IKKβ表达和NF-κB磷酸化，从而减缓乳腺癌细胞增殖，加速细胞凋亡，限制瘤体发育，从而缓解疾病。

Objective To explore the mechanism of lentinan combined with cisplatin on breast cancer by regulating IKKβ/NF-κB pathway. Methods Lentinan Capsules were prepared into 10, 20, 40 μg/mL solutions using complete medium, and cisplatin was prepared into 10 μmol/L solution. RPMI-1640 medium containing 10% fetal bovine serum was used for cell culture, and divided into model group, cisplatin group (10 μmol/L). Lentinan + cisplatin groups were treated with 10, 20, and 40 μg/mL lentinan on the basis of cisplatin group, respectively, and cultured for 48 h for in vitro cell experiment. CCK-8 assay was used to detect the proliferation of breast cancer MCF-7 cells, and the inhibition rate of cell proliferation was calculated. Cell apoptosis was detected by flow cytometry. The rats were randomly divided into model group, cisplatin (2 mg/kg) group, lentinan group (15 mg/kg), cisplatin (2 mg/kg) + lentinan (15 mg/kg) group, and Prostratin (0.5 mg/kg) group, with 10 rats in each group. The cultured breast cancer cells were successively injected into the mammary pad of each group for modeling. The rats in each group were given corresponding drug treatment, ip once daily, rats in model group were ip normal saline for 28 days. The tumor volume, quantity, and weight of each group were calculated. The expression levels of NF-κB p65, NF-κB p-p65, and IKKβ were detected by Western blotting. Results Compared with the model group, the proliferation inhibition rate and apoptosis rate of breast cancer cells in cisplatin group and cisplatin + lentinan (10, 20, 40 μg/mL) groups were significantly increased (P < 0.05). Compared with cisplatin group, the proliferation inhibition rate and apoptosis rate of cisplatin + lentinan (10, 20, 40 μg/mL) group were significantly increased (P < 0.05), and increased in a dose-dependent manner. Compared with the model group, the tumor volume, tumor weight, tumor number, the protein expression of NF-κB p-p65/NF-κB p65 and IKKβ in the cisplatin group, lentinan group, and cisplatin + lentinan group were significantly decreased (P < 0.05). Compared with the cisplatin group and lentinan group, the tumor volume, tumor weight, tumor number, NF-κB P-p65/NF-κB p65, and IKKβ protein expression in the cisplatin + lentinan group were significantly decreased (P < 0.05). Compared with the cisplatin + lentinan group, the tumor volume, tumor weight, tumor number, and protein expression levels of NF-κB P-p65/NF-κB p65 and IKKβ in Prostratin group were significantly increased (P < 0.05). Conclusion Cisplatin combined with lentinan can inhibit the expression of IKKβ and phosphorylation of NF-κB by regulating IKKβ/NF-κB pathway, thereby slowing down the proliferation of breast cancer cells, accelerating cell apoptosis, limiting tumor development and alleviating the disease.

R966

国家自然科学基金资助项目（81503392）