藉由增加TNF receptor-1表現Genistein促進 trichostatin A 抑制 A549細胞生長之效果

Genistein enhanced the effects of trichostatin A on inhibition of A549 cells growth by increasing the expression of TNF receptor-1.

楊盈芝(Ying-Chihi Yang)

指導教授 ： 葉姝蘭

中山醫學大學/健康管理學院/營養學研究所/碩士(2010年)

https://doi.org/10.6834/CSMU.2010.00134

摘要

我們先前的研究發現，genistein 促進trichostatin A （TSA）誘發
肺癌細胞A549 凋亡，但genistein 造成此結果的真正機制並不清楚，我們由microarray 分析結果發現可能與genistein 增加death receptor，tumor necrosis factor receptor-1 (TNFR-1) 的表現量有關，因此本研究主要目的是驗證genistein 增加TSA 抑制癌細胞生長是否透過調節TNFR-1，進而引起下游signaling cascade，而增加肺癌細胞A549 的凋亡。A549 細胞以TSA (50 ng) 和genistein (5、10μM) 單獨或合併處理一段時間後發現，如預期的， 5 和10 μM genistein 可顯著增加TSA 誘發細胞生長停滯，並具劑量效應，同時TSA 合併genistein 處理6 及12 小時，TNFR-1 的mRNA 和蛋白質的表現量亦顯著較TSA單獨處理組高，合併處理後，與TSA 單獨組相比，亦會增加caspase-10及caspase-3 mRNA 表現及活性，而caspase-8 mRNA 表現及活性則不受影響。我們利用轉染siRNA 的方式抑制A549 細胞內TNFR-1 的表現，結果發現genistein 促進TSA 抑制細胞生長及增加caspase-3 的活性的效應都被抑制。綜合以上結果證明，增加細胞凋亡外在途徑，亦即TNFR-1 表現，在genistein 促進TSA 誘發肺癌細胞A549 凋亡的機制上應扮演重要角色。

關鍵字

A549細胞 ； 金雀異黃酮 ； 曲古抑菌素 ； 腫瘤壞死因子接受器-1

並列摘要

Our previous study has shown that genistein enhance the apoptosis in A549 lung cancer cells induced by trichostatin Our previous study has shown that genistein enhance the apoptosis in A549 lung cancer cells induced by trichostatin A (TSA). The precise mechanisms underlying such an effect of genistein, however, is unclear.From microarray assay, we found that the increase of death receptor,tumor necrosis factor receptor-1 (TNFR-1), induced by genistein may
play an important role. Thus, in the present study, we investigated whether genistein enhance the anti-cancer effect of TSA through up-regulation of TNFR-1 death receptor signaling. We incubated A549 cells with TSA (50 ng/mL) alone or in combination with genistein for a
period of time. As expected, 5 and 10μM of genistein significantly increased the cell growth arrest induced by TSA in a time- and dose-dependent manner. Incubation of TSA in combination with genistein, TNFR-1 mRNA and protein expression were significantly increased at 6 and 12 hrs, respectively, as compared with that of TSA alone group. The combinative treatment also increased the mRNA expression and the activity of TSA-induced caspase-10, but not caspase-8,in A549 cells compared with TSA alone group. Furthermore, the combinative treatment increased the mRNA expression and the activity of TSA-induced caspase-3. Using transfection of siRNA to silence the expression of TNFR-1, we found that the enhancing effects of genistein on TSA-induced apoptosis, the caspase-3 activity in A549 cells were
suppressed. These data demonstrated that the up-regulation of TNFR-1 death receptor pathway play an important role, at least in part, in the enhancing effect of genistein on TSA-induced apoptosis in A549 cells.

並列關鍵字

A549 cell ； genistein ； trichostatin A ； tumor necrosis factor receptor-1

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