肺癌是全球死亡的主要原因之一。 PTEN 被廣泛認為是一種腫瘤抑制基因，在各種癌症類型中都表現出異常功能。在這項研究中，我們希望澄清PTEN 對肺癌進展的影響。首先，我們通過腹腔注射構建了肺癌 PTEN 敲除小鼠模型，該模型刪除了 EGFRL858R/PTEN-/-小鼠的 PTEN 外顯子 5（ EGFRL858R/PTEN-/- ）。結果表明， 與僅誘導 EGFRL858R相比 EGFRL858R/PTEN-/-小鼠觀察到明顯的支氣管增生，表明 PTEN 可能是負調控支氣管進展的。我們分析了下一代測序（ NGS） 數據，並發現 PTEN的喪失不僅增加了支氣管基因標記物的表達，還增加了與纖毛生成相關的基因的表達。另一方面， PTEN 的敲除導致 MUC5AC 的過度表達。其次，下調的 PTEN 活化核受體亞家族-NR2F1，一種纖毛基因的錄因因子，通過增加 AKT 的磷酸化。使用慢病毒-shNR2F1 敲低 NR2F1 的表達顯著降低了控制纖毛形成的一些基因： DNAI2， DNAI3。敲低 PTEN 還誘導了DNAI2 啟動子活性，表明 NR2F1 確實調控了 DNAI2 的表達。此外，我們發現 PTEN 在 PC9 (EGFRL858R)和 H1299L858R 細胞株中增加了AKT 的磷酸化，但在 A549 KRAS 突變和 H1299 細胞中降低了 AKT 的磷酸化。這些結果暗示 PTEN 在具有 EGFR 突變的肺癌中可能作為一個致癌基因，在具有 KRAS 突變的肺癌中可能作為一個腫瘤抑制基因。

Lung cancer is the leading cause of death globally. The survival percentage in lung cancer patients has been reported and poor prognosis due to metastasis after surgery. Cancer formation is caused by the activating of oncogenes or downregulating of tumor suppressors. PTEN was well known as a tumor suppressor, which is an abnormal function in various cancer types. In this study, we wanted to clarify the impact of PTEN on lung cancer progression. First, we constructed PTEN knock-out mice modal in lung cancer by intraperitoneal (IP) injections, which removed exon 5 of PTEN in lung cancer mice (EGFRL858R/PTEN-/-). The results indicated that significant bronchiolar hyperplasia was observed in EGFRL858R/PTEN-/- mice compared to EGFRL858R induced only, suggesting that PTEN may be negative bronchial progression. Next, we analyzed Next Generation Sequencing (NGS) data. We identified that loss of PTEN not only increased the expression of bronchial gene markers but also of genes regulated to ciliogenesis. In addition, PTEN downregulation-induced acetylated tubulin, a cilia marker was examined by immunohistochemistry (IHC). Through the development of cilia, we proposed that loss of PTEN contributes to lung cancer progression via the proliferation of bronchial cells. On the other hand, the elimination of PTEN resulted in the overexpression of MUC5AC, a critical component of lung defense studied by IHC and Alcian Blue (AB) staining. Secondly, downregulated PTEN activated NR2F1, a transcription factor for cilia genes through increasing phosphorylation of AKT. Knockdown of the expression of NR2F1 by using lentivirus-shNR2F1 led to a dramatic decrease in certain genes that control cilia formation: DNAI2, and DNAI3. Deletion of PTEN also induced DNAI2 promoter activity suggesting that NR2F1 regulated DNAI2 expression. Moreover, we discovered that PTEN elevated phosphor AKT in PC9 (EGFRL858R), and H1299L858R cells but decreased it in A549 (KRAS mutant), and H1299 cells. PTEN functions as a double-edged sword, regulating the development of lung cancer in different genome backgrounds. By elucidating this detailed mechanism, we hope that we can provide beneficial diagnosis and therapeutic strategies for cancer treatment.

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