大腸結腸癌在世界上患病率排名為前三名，年齡越高則有更高機率罹患此疾病。目前奧沙利鉑(Oxaliplatin)為主要治療大腸結腸癌藥物之一，但患者在經過長期服藥後便會產生抗藥性，目前對於癌症抗藥機制至今還未清楚，所以針對其治療方法至今仍須不斷深入探討。在近幾年之細胞培養方法中，發現3D立體細胞培養可透過胞外基質 (extracellular matrix; ECM)微環境，更趨近人體內組織結構。近年，發現外泌體中微核糖核酸(exosomal microRNAs)有特定方向性的癌症轉移，以及抗藥性相關分子機制。因此，本研究針對在2D和3D 大腸結腸癌細胞以及抗藥細胞所分泌出之外泌體，進行高通量定序來比較兩者間miRNA的表現差異。我們發現miR-182-5p、miR-200a-3p、miR-10a-5p在2D和3D的外泌體中有明顯之差異表達現象。並且進一步實驗結果也驗證miRNA在大腸結腸癌細胞中的表現，確定miR-182-5p、miR-200a-3p的表現會增加。在過表達中miR-200a-3p進入結腸癌細胞後發現細胞存活有特異性之表達。最後，經由miR-200a-3p的表達結果，期望未來可以利用這些miRNAs作為大腸結腸癌抗藥的生物標誌物以作為治療癌症的重要角色。

Colorectal cancer (CRC) poses a significant global health challenge, compounded by the complexity of chemotherapy resistance in treatment strategies. Conventional two-dimensional (2D) cell culture models have inherent limitations in replicating the intricate tumor microenvironment. In contrast, three-dimensional (3D) models offer superior mimicry of in vivo conditions. This study focuses on investigating miRNA expression profiles within exosomes derived from both 2D and 3D cultures of HCT116 cells and their drug-resistant variants (Oxa#7, Oxa#9). Through comprehensive miRNA sequencing, we have identified distinct patterns of miRNA expression, particularly highlighting miR-182-5p, miR-10a-5p, miR-200a-3p, miR-98-5p and miR-185-5p which may elucidate critical mechanisms underpinning drug resistance in CRC. Further qPCR experiments confirmed significant upregulation of miR-182-5p, miR-200a-3p in exosomes from drug-resistant cells, suggesting their potential key roles in drug resistance mechanisms. Functional assays demonstrated that transfection with miR-200a-3p mimics notably enhanced sensitivity of HCT116 cells to oxaliplatin treatment. These findings indicate that upregulation of miR-200a-3p could potentially overcome oxaliplatin resistance and enhance cancer cell sensitivity to therapy, implicating them as promising targets for combating drug resistance in CRC. This research contributes to advancing our understanding of how 3D culture systems can more accurately simulate the complexities of CRC biology and therapeutic responses, providing a theoretical foundation for further development of treatment strategies targeting chemotherapy resistance.

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