[關(guān)鍵詞]
[摘要]
目的 探究香菇多糖聯(lián)合順鉑通過(guò)調(diào)控I-κB激酶β(IKKβ)/核因子-κB(NF-κB)通路對(duì)乳腺癌的作用機(jī)制。方法 采用完全培養(yǎng)基將香菇多糖膠囊制備成10、20、40 μg/mL的溶液,將順鉑制備成10 μmol/L溶液。選用含有10%胎牛血清的RPMI-1640培養(yǎng)液進(jìn)行細(xì)胞培養(yǎng),將其分為模型組、順鉑組(10 μmol/L),順鉑+香菇多糖組在順鉑組處理的基礎(chǔ)上,分別加10、20、40 μg/mL香菇多糖,培養(yǎng)48 h用于體外細(xì)胞實(shí)驗(yàn),CCK-8法檢測(cè)乳腺癌MCF-7細(xì)胞增殖情況,計(jì)算細(xì)胞增殖抑制率;流式細(xì)胞術(shù)檢測(cè)細(xì)胞凋亡情況。隨機(jī)將大鼠分為模型組、順鉑組(2 mg/kg)、香菇多糖組(15 mg/kg)、順鉑(2 mg/kg)+香菇多糖(15 mg/kg)組、Prostratin(0.5 mg/kg)組,每組10只。將培養(yǎng)的乳腺癌細(xì)胞依次注射于各組大鼠乳腺墊內(nèi)進(jìn)行造模。各組大鼠進(jìn)行相應(yīng)的給藥處理,每日ip 1次,模型組大鼠ip等劑量的生理鹽水,連續(xù)干預(yù)28 d。計(jì)算各組大鼠瘤體體積、數(shù)量并稱重;Western blotting法檢測(cè)各組大鼠NF-κB p65、NF-κB p-p65、IKKβ表達(dá)水平。結(jié)果 與模型組相比,順鉑組、順鉑+香菇多糖(10、20、40 μg/mL)組乳腺癌細(xì)胞增殖抑制率、細(xì)胞凋亡率顯著升高(P<0.05);與順鉑組相比,順鉑+香菇多糖(10、20、40 μg/mL)乳腺癌細(xì)胞增殖抑制率、細(xì)胞凋亡率顯著升高(P<0.05),且呈劑量相關(guān)性增長(zhǎng)。與模型組相比,順鉑組、香菇多糖組、順鉑+香菇多糖組大鼠瘤體體積、瘤體質(zhì)量、瘤體數(shù)目、瘤體內(nèi)NF-κB p-p65/NF-κB p65、IKKβ蛋白表達(dá)量顯著降低(P<0.05);與順鉑組、香菇多糖組相比,順鉑+香菇多糖組大鼠瘤體體積、瘤體質(zhì)量、瘤體數(shù)目、瘤體內(nèi)NF-κB p-p65/NF-κB p65、IKKβ蛋白表達(dá)量顯著降低(P<0.05);與順鉑+香菇多糖組相比,Prostratin組大鼠瘤體體積、瘤體質(zhì)量、瘤體數(shù)目、瘤體內(nèi)NF-κB p-p65/NF-κB p65、IKKβ蛋白表達(dá)量顯著升高(P<0.05)。結(jié)論 順鉑聯(lián)合香菇多糖可通過(guò)調(diào)控IKKβ/NF-κB通路,抑制IKKβ表達(dá)和NF-κB磷酸化,從而減緩乳腺癌細(xì)胞增殖,加速細(xì)胞凋亡,限制瘤體發(fā)育,從而緩解疾病。
[Key word]
[Abstract]
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.
[中圖分類(lèi)號(hào)]
R966
[基金項(xiàng)目]
國(guó)家自然科學(xué)基金資助項(xiàng)目(81503392)