[關(guān)鍵詞]
[摘要]
目的 探討丹酚酸B對結(jié)腸癌細(xì)胞HCT116增殖、遷移、糖酵解和上皮-間充質(zhì)轉(zhuǎn)化(EMT)的作用及機(jī)制。方法 通過CCK-8實(shí)驗(yàn)和克隆形成實(shí)驗(yàn)評估丹酚酸B(25、50、100 μmol·L-1)對結(jié)腸癌細(xì)胞HCT116增殖的影響;通過創(chuàng)傷愈合實(shí)驗(yàn)評估丹酚酸B對HCT116細(xì)胞遷移能力的影響;試劑盒法檢測丹酚酸B對結(jié)腸癌細(xì)胞糖酵解及乳酸水平的影響;通過Western blotting法分析丹酚酸B對EMT相關(guān)蛋白E-鈣黏蛋白(E-cadherin)、N-鈣黏蛋白(N-cadherin)、波形蛋白(Vimentin)和Snail,糖酵解相關(guān)蛋白葡萄糖轉(zhuǎn)運(yùn)蛋白1 (GLUT1)、乳酸脫氫酶A (LDHA),p-絲氨酸/蘇氨酸激酶(AKT)/核因子κB(NF-κB)信號(hào)通路相關(guān)蛋白AKT、p-p65的調(diào)控作用。結(jié)果 與對照組相比,50、100 μmol·L-1丹酚酸B顯著抑制HCT116細(xì)胞的增殖(P<0.01、0.001);100 μmol·L-1丹酚酸B顯著阻斷HCT116細(xì)胞的遷移(P<0.001); 50、100 μmol·L-1丹酚酸B顯著降低HCT116細(xì)胞GLUT1和LDHA的蛋白表達(dá)(P<0.01、0.001),顯著降低HCT116細(xì)胞的葡萄糖消耗及乳酸產(chǎn)生(P<0.01、0.001);50、100 μmol·L-1丹酚酸B組上皮標(biāo)志物E-cadherin的蛋白表達(dá)顯著上升(P<0.01、0.001),而間充質(zhì)標(biāo)志物N-cadherin、Vimentin和轉(zhuǎn)錄因子Snail的蛋白表達(dá)顯著下降(P<0.01、0.001);50、100 μmol·L-1丹酚酸B顯著降低AKT的磷酸化水平和核因子κB(NF-κB)的亞基p65的磷酸化水平(P<0.01、0.001)。結(jié)論 丹酚酸B能顯著抑制結(jié)腸癌細(xì)胞的增殖、遷移能力,并有效抑制EMT過程、糖酵解,可能通過抑制AKT/NF-κB信號(hào)通路實(shí)現(xiàn)。
[Key word]
[Abstract]
Objective To investigate the effects of salvianolic acid B on the proliferation, migration, glycolysis, and epithelialmesenchymal transition (EMT) of colon cancer cells HCT116 and the related mechanism. Methods The proliferation of HCT116 cells was evaluated by CCK-8 assay and colony formation assay with salvianolic acid B (25, 50, and 100 μmol·L-1). The migration ability of HCT116 cells was evaluated by wound healing assay with salvianolic acid B. The glycolysis and lactate levels of HCT116 cells were detected by kit assay. The effects of salvianolic acid B on the EMT-related proteins E-cadherin, N-cadherin, Vimentin, and Snail, the glycolysis-related proteins GLUT1 and LDHA, and the p-serine/threonine kinase (AKT)/nuclear factor kappa B (NF-κB) signaling pathway-related proteins AKT and p-p65 were analyzed by Western blotting. Results Compared with the control group, 50 and 100 μmol·L-1 salvianolic acid B significantly inhibited the proliferation of HCT116 cells (P < 0.01, 0.001), 100 μmol·L-1 salvianolic acid B significantly blocked the migration of HCT116 cells (P < 0.001), 50 and 100 μmol·L-1 salvianolic acid B significantly reduced the protein expression of GLUT1 and LDHA in HCT116 cells (P < 0.01, 0.001), and significantly reduced the consumption of glucose and lactate in HCT116 cells (P < 0.01, 0.001). 50 and 100 μmol·L-1 of salvianolic acid B treatment resulted in a significant increase in the protein expression of epithelial marker E-cadherin (P < 0.01, 0.001), while the expression of mesenchymal markers N-cadherin, Vimentin, and transcription factor Snail significantly decreased (P < 0.01, 0.001), 50 and 100 μmol·L-1 of salvianolic acid B significantly reduced the phosphorylation levels of AKT and the subunit p65 of NF-κB (P < 0.01, 0.001). Conclusion Salvianolic acid B can significantly inhibit the proliferation and migration of colon cancer cells, effectively inhibit the EMT process and glycolysis, and may achieve this by inhibiting the AKT/NF-κ B signaling pathway.
[中圖分類號(hào)]
R285.5
[基金項(xiàng)目]
國家自然科學(xué)基金資助項(xiàng)目(81660302)