初級纖毛在細胞訊息傳遞以及控制細胞週期進行中扮演著重要的作用。過去認為，當細胞週期停在G0期時，初級纖毛會開始生長。初級纖毛開始生長的時候，TTBK2會被傳送到母親中心粒的位置，然後將在母親中心粒上的CP110磷酸化。而磷酸化的CP110會從母親中心粒上離開，接著TTBK2會募集生長初級纖毛的相關蛋白使初級纖毛能夠開始生成。長期以來人們一直認為癌細胞並不會生長初級纖毛。然而，最近的研究表明，在有些癌細胞中，如肺腺癌A549細胞，在細胞骨架被破壞或接受外在壓力刺激時會長出初級纖毛的結構。所以在本實驗中，我們想研究抗癌藥物VP-16 (Etoposide)對於A549中心體的作用，並試圖找到出初級在其中所扮演的角色。
在VP-16 (Etoposide)處理後，A549細胞中並未觀察到中心體異常增生，相反的，我們觀察到了初級纖毛的生長。所以我們接著觀察初級纖毛生長是否受到DNA損傷反應的調控。觀察到上游相關蛋白如DNA-PK，ATM和ATR的活化，然而，在處理抑制劑後發現這些蛋白的活性抑制不影響初級纖毛生長。有趣的是，我們發現Akt的抑制會降低VP-16 (Etoposide)處理後的初級纖毛增生。所以Akt在VP-16 (Etoposide)誘發的初級纖毛生長中扮演著關鍵的作用。最近的研究顯示抑制PI3K / AKT訊息傳遞通路會促進細胞自噬，因此我們也觀察了細胞自噬作用。我們的結果顯示，當VP-16 (Etoposide)處理後肺腺癌A549細胞會啟動自噬作用。
最後我們想知道初級纖毛在A549細胞中所扮演的角色，因此我們使用siRNA敲除初級纖毛內的運輸蛋白並處理了VP-16 (Etoposide)以觀察有無初級纖毛的差異，結果顯示，在VP-16 (Etoposide)加入後，初級纖毛被敲除的組別細胞死亡率較高。根據我們的實驗結果，我們認為初級纖毛在VP-16 (Etoposide)處理後生長對於A549細胞是起一個保護的作用。
我們的實驗結果證實，VP-16 (Etoposide)加入會促進A549細胞初級纖毛形成，並且是透過Akt的影響。根據目前的實驗結果，初步認為初級纖毛在VP-16 (Etoposide)加入後對A549細胞起保護作用。在添加抗癌藥物後細胞促進其生長以維持細胞存活。

SUMMARY

The primary cilium extends outside the cell as a cellular antenna, and primary cilium was important in cell cycle progression. It has been suggested that cancer cells do not grow primary cilia. However, recent studies show that they may have the ability to grow cilia when stresses, but its biological significance is still unclear. We would like to study the effect of anticancer drug Etoposide on the centrosome of A549, and we founded that after Etoposide treatment for 24 hours, A549 induce cilia formation. Upon Etoposide treatment, DNA damage response up stream markers like ATM, ATR and DNA-PKcs didn’t affect the cilia formation in A549. Its seems that Etoposide were induced DNA damage response in A549 cells, but the mechanism in Etoposide induced A549 cells ciliogenesis are not go through the same way. Then we found that Akt was activated and co-treatment of Etoposide and Akt inhibitors, the primary cilia formation was reduced. In addition, we also found the autophagy was activated by Etoposide, and the activated autophagy facilitated ciliogenesis and maintained cell survival. To further confirm the role of primary cilia upon Etoposide treated, primary cilia were disrupted by siRNA against intraflagellar transport protein IFT88, and we observed reduced cell viability was observed in cilia deficient cells. Thus, Etoposide induced primary cilia for protecting A549 cells from death via Akt and/or autophagy pathways.

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