| 李 吉,徐 莹,褚以忞,蒯 榕,周 璐,张海芹.lncRNA XIST-miR137-ATG5调节细胞自噬功能在肠癌细胞5-氟胞嘧啶耐药性中的作用[J].,2020,(19):3609-3615 |
| lncRNA XIST-miR137-ATG5调节细胞自噬功能在肠癌细胞5-氟胞嘧啶耐药性中的作用 |
| LncRNA XIST-miR137-ATG5 Mediate Cell Autophagy to Attenuate 5-FU Resistance in Colorectal Cancer Cells to 5-FU |
| 投稿时间：2020-02-28 修订日期：2020-03-23 |
| DOI：10.13241/j.cnki.pmb.2020.19.002 |
| 中文关键词: microRNA XIST 结肠癌 细胞自噬 化疗敏感性 |
| 英文关键词: microRNA XIST Colorectal cancer Autophagy Chemoresistance |
| 基金项目:上海市卫生和计划生育委员会青年项目（20174Y0084）；上海交通大学医工（理）交叉基金项目（ZH2018QNB24） |
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| 中文摘要: |
| 摘要 目的：本文旨在研究长链非编码RNA XIST-miR137-ATG5的相互作用，同时探讨其调节细胞自噬功能与肠癌细胞5-氟胞嘧啶敏感性的关系。方法：实时聚合酶链反应(real time PCR)检测XIST与miR-137在肠癌细胞中的表达；采用脂质体转染法将si-XIST，miR-137转染入肠癌SW480及HCT116细胞中。采用CCK-8检测瞬时转染si-XIST对肠癌细胞增殖及5-FU敏感性的影响；并利用双荧光素酶报告实验检测miR-137与XIST, miR-137与ATG5相互关系。Western blot方法检测XIST- miR137- ATG5对细胞自噬的影响。结果：与正常结肠细胞FHC比较， XIST在结肠癌细胞系明显高表达，miR-137在结肠癌细胞系明显低表达。与阴性对照组比较，转染si-XIST后，SW480及HCT116细胞增殖能力明显受到抑制，对F-5U的敏感性增强，且抑制自噬蛋白Beclin-1及LC3II/LC3 I的表达。miR-137可与XIST，ATG5 3'UTR结合，抑制XIST和ATG5的表达及功能。在结肠癌SW480细胞中共转染miR-137 inhibitor或过表达ATG5可逆转XIST沉默引起的5-FU耐药，同时可逆转因XIST沉默引起的自噬蛋白表达的抑制。结论：LncRNA XIST或可通过调控mir137-ATG促进结直肠癌细胞SW480自噬从而提高其对5-FU的耐药，针对其这一机制，可为将来针对结肠癌的靶向治疗提供一定的实验基础。 |
| 英文摘要: |
| ABSTRACT Objective: To investigate the interaction of long noncoding RNA X-inactive specific transcript (XIST) -miR137-ATG5 and explore the relationship between its function of autophagy mediating and 5-FU sensitivity in colorectal cancer cells. Methods: The relative expressions of XIST and miR-137 in colorectal cancer cell lines were detected by real-time polymerase chain reaction(qRT-PCR). The lipofection assay was utilized in XIST and miR-137 transfection experiments. CCK-8 assay was used to detect cell viability and the 5-FU sensitivity in colorectal cancer cells. Moreover, luciferase reporter system was used to determine binding ability between miR-137 and XIST or ATG5. Additionally, indicating protein expressions were determined by western blotting. Results: We found that XIST expression was increased in CRC cells comparing to normal colon cell FHC, which showed a negative correlation with miR-137 expression. In addition, XIST knockdown noticeably inhibited the proliferation of CRC cells and enhanced 5-FU sensitivity. It revealed that XIST knockdown suppressed the expression of Beclin-1 and ratio of LC3II/I. miR-137 was validated to target XIST and 3'UTR of ATG5, playing an important role in restraining the expression and function of XIST and ATG5 . Inhibition of miR-137 or overexpression of ATG5 could effectively reverse elevated 5-FU sensitivity upon XIST knockdown in SW480. Conclusion: It is suggested that LncRNA XIST could target miR-137 promoting autophagy to facilitate 5-FU chemoresistance in CRC and XIST-miR137-ATG5 may serve as a potential therapeutic target for patients with colorectal cancers. |
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