本研究旨探討全氟及多氟烷基物質（per- and polyfluoroalkyl substances， PFAS）在促進化療抗藥性泌尿上皮癌（urothelial carcinoma， UC）惡性進展中的作用及其潛在分子機制。PFAS為廣泛存在於環境中的持久性污染物，具有高度穩定性與生物累積性，近年研究指出其暴露與多種癌症風險上升相關。然而，目前對於 PFAS 是否能進一步促進已具化療抗藥性之腫瘤細胞惡化，仍缺乏系統性研究。 本研究聚焦於長期低劑量PFAS暴露對抗藥性腫瘤行為之影響，並比較長鏈代表 物質全氟辛烷磺酸（PFOS）與短鏈替代物全氟丁烷磺酸（FBSA）之作用差異。 此實驗以人類膀胱癌細胞株 J82 及其經持續順鉑處理建立之抗藥性衍生株 J82 CR0.65 為實驗模型。相較於親代細胞，J82-CR0.65 細胞呈現顯著提升之遷移能 力與基質金屬蛋白酶（matrix metalloproteinases， MMPs）表現，顯示其具備較強之惡性特徵。在長期低劑量PFOS暴露條件下，兩種細胞株皆出現細胞存活率與 集落形成能力上升，其中以抗藥性細胞 J82-CR0.65 的變化更為顯著，顯示抗藥 性腫瘤細胞對 PFAS 誘導之惡性表型具有較高敏感性。進一步分析顯示，PFOS 暴露可隨時間促進上皮–間質轉換（epithelial–mesenchymal transition， EMT）及細胞外基質（extracellular matrix， ECM）重塑，並伴隨 MMP-2、MMP-7及MMP-9等分子表現上調。此外，在親代細胞中，短期（7天）PFOS暴露已可誘導EMT相關蛋白（如N-cadherin、Snail 及 fibronectin）表現增加，並使其 MMP 表現趨近於抗藥性細胞之型態；而在抗藥性細胞中，長期（約30天）PFOS暴露則主要誘導發炎相關訊號分子，包括IL-1β、IL-18及KRAS之上升。在抗藥性細胞中， 上述發炎與EMT變化並未伴隨顯著之活性氧（reactive oxygen species， ROS）變 化，顯示PFOS可能透過非典型氧化壓力途徑，促進EMT與發炎訊號之交互作 用（EMT–inflammatory axis）， 進而驅動腫瘤惡性進展。此外，短鏈PFAS替代物FBSA 亦呈現類似效應，包括誘導 EMT 相關分子、MMP 表現及趨化激素（如CXCL5、CXCL9）轉錄上升，顯示即使為替代性PFAS，仍可能保留其促腫瘤進 展之潛在能力。綜合以上結果，本研究指出長期PFAS暴露可透過協同活化EMT、 細胞外基質重塑及發炎訊號途徑，促進順鉑抗藥性泌尿上皮癌細胞之惡性進展， 且抗藥性細胞對環境污染物具有較高反應性。本研究不僅補足PFAS於抗藥性腫 瘤進展中的角色，也進一步強調環境暴露因子在腫瘤惡化過程中的重要性，對未 來癌症治療策略與環境風險評估具有潛在意義。;This study aimed to investigate the role of per- and polyfluoroalkyl substances (PFAS) in promoting malignant progression in chemoresistant urothelial carcinoma (UC) and to elucidate the underlying molecular mechanisms. PFAS are persistent environmental pollutants characterized by high chemical stability and bioaccumulation， and increasing evidence has linked their exposure to elevated cancer risk. However， whether PFAS further exacerbate malignant phenotypes in chemoresistant tumor cells remains largely unclear. Therefore， this study focused on the effects of long-term， low-dose PFAS exposure on chemoresistant cancer cells， with a comparative analysis of the long-chain compound perfluorooctane sulfonate (PFOS) and the short-chain alternative perfluorobutane sulfonate (FBSA). Human bladder cancer J82 cells and their cisplatin-resistant derivative， J82-CR0.65， were used as experimental models. Compared with parental cells， J82-CR0.65 cells exhibited enhanced migratory capacity and elevated expression of matrix metalloproteinases (MMPs)， indicating a more aggressive phenotype. Under long-term， low-dose PFOS exposure， both cell lines showed increased cell viability and clonogenic potential， with more pronounced effects observed in the resistant cells， suggesting that chemoresistant cells are more susceptible to PFAS-induced malignant progression.Further analyses revealed that PFOS exposure promoted epithelial–mesenchymal transition (EMT) and extracellular matrix (ECM) remodeling in a time-dependent manner， accompanied by upregulation of MMP-2， MMP-7， and MMP-9. In parental J82 cells， short-term PFOS exposure (7 days) induced EMT-related markers， including N-cadherin， Snail， and fibronectin， and shifted the MMP expression profile toward that observed in resistant cells. In contrast， prolonged PFOS exposure (approximately 30 days) in J82-CR0.65 cells predominantly activated inflammatory signaling pathways， as evidenced by increased expression of IL-1β， IL-18， and KRAS. Notably， these inflammatory and EMT-associated changes occurred without significant alterations in intracellular reactive oxygen species (ROS)， suggesting that PFOS may promote malignant progression through a noncanonical mechanism involving crosstalk between EMT and inflammatory signaling (EMT–inflammatory axis)， rather than through classical oxidative stress pathways. The short-chain PFAS alternative FBSA elicited comparable effects， including induction of EMT markers， MMP expression， and chemokine transcription (e.g.， CXCL5 and CXCL9)， indicating that replacement PFAS may retain pro-tumorigenic potential. In conclusion， long-term PFAS exposure promotes malignant progression in cisplatin-resistant urothelial carcinoma through coordinated activation of EMT， extracellular matrix remodeling， and inflammatory signaling pathways. These findings suggest that chemoresistant tumor cells exhibit heightened responsiveness to environmental stressors such as PFAS， and highlight the potential contribution of environmental exposure to tumor progression and therapeutic resistance.
