蜜環菌萃出的sesquiterpene aryl ester抑制鎳誘發A549及H1975細胞侵犯及轉移經由向下調節TLR4/NF-kB訊號

A sesquiterpene aryl ester from Armillaria mellea suppresses the promoting effect of nickel-induced invasion and migration in A549 and H1975 cells through downregulation of TLR4/NF-kB signaling

余佩璇(Pei-syuan Yu)

指導教授 ： 葉姝蘭 ； 劉凱莉 ； 陳建志

中山醫學大學/健康管理學院/營養學系碩士班/碩士(2014年)

https://doi.org/10.6834/CSMU.2014.00185

摘要

研究顯示，鎳是一種普遍存在於環境當中的污染物，會經由TLR4信號傳遞路徑促進肺癌細胞的侵犯能力。我們先前的研究發現，蜜環菌中萃取的一種化合物sesquiterpene aryl ester (melleolide Q; AM-Q)能顯著抑制鎳誘發肺癌細胞A549、H1975 (EGFR突變)細胞的侵犯及體外轉移能力，然而，其可能的機制目前還不清楚。蜜環菌屬於口蘑科家族，為一種可食用的蘑菇，在中醫裡蜜環菌已被用於治療老年人頭痛和頭暈。在本研究中，我們探討AM-Q是否能經由調控TLR4/NF-κB訊號來抑制鎳所造成的影響。因此，本研究先以A549及H1975細胞分別與R (2或5μM)、CLI-095 (TLR4 inhibitor, 3μM)或SC-514 (IKK inhibitor, 100μM)預培養後，再以鎳 (NiCl2, 1 mM)培養一段時間，之後再進行各項分析。結果發現，抑制劑CLI-095及SC-514能顯著抑制鎳誘發A549細胞的侵犯及轉移能力，而鎳暴露會顯著增加A549細胞TLR4/Myd88蛋白的表現、IκB kinase (IKK)的磷酸化作用、進而促進p65進入核內，增加MMP-9活性的表現。而AM-Q能抑制鎳誘發A549細胞的各種影響，其效果與抑制劑相似。另外，我們也發現，此兩種抑制劑也會抑制H1975細胞的侵犯及轉移能力，其效果與A549細胞一致﹔AM-Q也會抑制鎳誘發H1975細胞的Myd88、IκB kinase (IKK)的磷酸化作用、進而促進p65進入核內。綜合上述結果，本研究顯示AM-Q可以經由向下調控TLR4/NF-κB訊號傳遞路徑，抑制鎳造成肺癌細胞侵犯及轉移能力。

關鍵字

鎳 ； 蜜環菌 ； AM-Q

並列摘要

Studies showed that nickel, a pollutant commonly found in environment, exposure promotes the invasive potential of human lung cancer cells through TLR4 signaling. Our previous study found that a sesquiterpene aryl ester (melleolide Q; AM-Q), a compound extracted from Armillaria mellea, significantly inhibited Ni-induced invasion and migration in human lung cancer cell lines A549 and H1975 (EGFR mutation). However the possible mechanisms are unclear. Armillaria mellea, from the Tricholomataceae family, is an edible mushroom. It has been traditionally used in Chinese medicine to treat elderly condition including headache and dizziness. In the present study, we investigated whether AM-Q inhibit the effects of Ni through the regulation of TLR4/ NF-κB signaling. Human lung cancer A549 cells and H1975 cells were pre-incubated with AM-Q (2 or 5μM), CLI-095 (TLR4 inhibitor, 3μM) or SC-514 (IKK inhibitor, 100μM). Then, the cells were treated with NiCl2 (1 mM). First we found that the inhibitors, CLI-095 and SC-514, significantly inhibited the invasion and migration in Ni-treated A549 cells. Ni-exposure significantly increased the mRNA and protein expression of TLR4 and Myd88, the phosphorylation of IκB kinase (IKK), the translocation of p65 and the MMP-9 expression and activation in A549 cells. AM-Q suppressed all the effects of Ni in A549 cells. The effects of AM-Q were comparable to those of inhibitors. Furthermore, we found that the two inhibitors also inhibited the invasion and migration in H1975 cells. In addition, in consistent with the findings in A549 cells, AM-Q also tended to inhibit the expression of Myd88, the phosphorylation IKKβand the translocation of NF-κB induced by Ni-exposure in H1975 cells. In conclusion, these results indicate that the AM-Q may inhibit Ni-promoted lung cancer cell invasion and migration through downregulation of TLR4/NF-κB signaling, at least in part.

並列關鍵字

sesquiterpene aryl ester ； A549 ； H1975 ； nickel

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