肺癌為全球死亡率最高的癌症，去泛素化酶移除目標底物上的泛素以調節蛋白質降解，過去實驗室的研究指出泛素特異性蛋白酶24 ( ubiquitin-specific peptidase 24, USP24) USP24在體外肺癌細胞中抑制細胞凋亡並增加細胞增殖，穩定含溴結構域的蛋白質使晚期肺癌惡化，以及增加ABC轉運蛋白 P-gp、ABCG2 的含量，增強耐藥性，但對於USP24在肺癌初期進展中的確切角色需進一步釐清。在實驗室的動物實驗發現，Doxycycline induced- EGFRL858R*USP24C1695A小鼠相比EGFRL858R*USP24WT小鼠顯著抑制肺部腫瘤生長，這表明USP24作為致癌基因在體內促進癌症的形成，進而對USP24功能性缺失的小鼠之肺臟進行次世代RNA定序分析，顯示無論有無doxycycline誘導的小鼠，USP24缺失都高度影響免疫反應的啟動、體液免疫反應、免疫球蛋白、補體系統的基因表達，也在脾臟中看到B細胞數量增加，據此推論USP24可能在肺癌初期進展期間參與B細胞的調節。在RAMOS B細胞中，以USP24抑制劑處理顯著提升CLEC10A、MNDAL B細胞抗原相關基因以及PAX5 B細胞發育基因，降低USP24表現量也使CD79a、CD79b、IGHV1-9、IGKV14-111、IGHM 等B細胞受體與抗體基因表現上升，上調NFκB路徑活性，腫瘤浸潤性B細胞（tumor-infiltrating B cell，TIB）在調節癌症也有重要的協同作用，RAMOS細胞培養於PC9 USP24KD 的條件培養液( conditioned medium, CM)，使B細胞增殖能力與遷移能力提升，上調CLEC10A啟動發炎反應，細胞受體與抗體基因表現上升。本研究的結論顯示：在癌症形成過程中，USP24在B細胞中會高度表現，進而抑制B細胞的免疫活性，導致癌症細胞得以逃脫免疫系統的毒殺，而USP24的抑制劑（USP24-i-101）可以活化B細胞的免疫反應，增進免疫治療的效果。

Lung cancer is the highest cause of cancer-related deaths worldwide. USP24 is a deubiquitinating enzyme (DUB) that participates in regulating protein degradation in cells. This process mediates of ubiquitination levels in target proteins. Previous studies in our lab have indicated that ubiquitin-specific peptidase 24 (USP24) inhibits cancer cell apoptosis, and enhances cell proliferation in vitro. Also, it promotes lung cancer by stabilizing proteins p300 and β-TrCP in the tumor microenvironment, contributing to the progression and deterioration of late-stage lung cancer. Therefore, the exact role of USP24 in lung cancer progression in vivo requires further clarification. In previous laboratory experiments, doxycycline-induced EGFRL858RUSP24C1695A mice significantly suppressed lung tumor growth compared to EGFRL858RUSP24WT mice, suggesting that USP24 acts as an oncogene in promoting cancer formation in vivo. Subsequent RNA sequencing analysis of USP24C1695A mouse lungs, regardless of doxycycline induction, showed substantial impacts on the activation of immune responses, humoral immune response, immunoglobulins, and the complement system. Additionally, an increase in B cell numbers in the spleen was observed, implying USP24 may be involved in the regulation of B cells during cancer progression. Treatment with USP24 inhibitor in RAMOS B cells significantly upregulated the expression of B cell antigen-related genes (CLEC10A, MNDAL) and B cell development genes (PAX5), while reducing USP24 expression resulted in increased expression of B cell receptor and antibody genes (CD79a, CD79b, IGHV1-9, IGKV14-111, IGHM), meanwhile enhanced NFκB pathway activity. Tumor-infiltrating B cells (TIB) play a crucial synergistic role in regulating cancer. Culturing RAMOS cells in conditioned medium (CM) from PC9 USP24KD cells enhanced B cell proliferation and migration, triggered inflammatory responses via upregulation of CLEC10A, and increased expression of B cell receptor and antibody genes. In summary, USP24 express in B cells during cancer progression to inhibit B cell activity, leading cancer growth and malignancy. USP24-i-101 targeting USP24 increases B cell activity, subsequently inhibiting cancer progression.

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