南方医科大学学报

南方医科大学学报 ›› 2019, Vol. 39 ›› Issue (03): 357-.doi: 10.12122/j.issn.1673-4254.2019.03.15

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长片段非编码RNA XIST可改变人鼻咽癌HNE1 细胞对顺铂的耐药性

王浩，李维，谭国林   

  • 出版日期:2019-03-20 发布日期:2019-03-20

Long non-coding RNA XIST modulates cisplatin resistance by altering PDCD4 and Fas-L expressions in human nasopharyngeal carcinoma HNE1 cells in vitro

  • Online:2019-03-20 Published:2019-03-20

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摘要： 目的明确长片段非编码RNA（lncRNA）X失活特异性转录本（XIST）对人鼻咽癌细胞顺铂（DDP）耐药性的影响及可能的 机制。方法通过荧光定量PCR检测人鼻咽癌顺铂耐药HNE1细胞株（HNE1/DDP）中XIST的表达特征。采用MTT试验观察 XIST上调和下调对HNE1/DDP细胞DDP耐药性的影响，并通过EdU试验和流式细胞技术观察其对HNE1/DDP细胞增殖和凋 亡的影响。采用蛋白质印迹法检测XIST上调和下调对促凋亡因子程序性细胞死亡4（PDCD4）和Fas配体（Fas-L）蛋白表达的 影响。结果XIST在HNE1/DDP细胞的表达量显著高于HNE1细胞（0.57±0.06 vs 0.1±0.02，P<0.05）。下调XIST的表达显著降 低HNE1/DDP 细胞对DDP 的耐药性（P<0.05），而上调XIST 增加对DDP 的耐药性（P<0.05）。下调XIST 的表达显著降低 HNE1/DDP细胞增殖（6.17±1.93 vs 16.59 4.86, P<0.05）并诱导细胞凋亡[（18.04±4.72）% vs（4.22±1.65）%, P<0.05]，而上调XIST 增加细胞增殖（25.40±7.21 vs 13.16±3.95, P<0.05）并抑制细胞凋亡[（2.82±0.88）% vs（6.46±1.75）%, P<0.05]。下调XIST的表达 显著增加HNE1/DDP细胞中PDCD4（4.10±0.42 vs 1.0±0.12, P<0.05和Fas-L（3.07 0.29 vs 1.0±0.14, P<0.05）蛋白的表达，而上调 XIST降低PDCD4（0.36±0.11 vs 1.0±0.18, P<0.05）和Fas-L（0.17±0.09 vs 1.0±0.21, P<0.05）蛋白的表达。结论XIST在HNE1/ DDP细胞中表达升高，下调和上调XIST的表达可分别降低和增加HNE1/DDP细胞对DDP的耐药性，其机制可能与PDCD4和 Fas-L蛋白表达改变相关。

Abstract: Objective To explore the role of long non-coding RNA (lncRNA) X inactive specific transcript (XIST) in modulating cisplatin (DDP) resistance of human nasopharyngeal carcinoma cells and investigate the possible mechanism. Methods Realtime PCR was performed to detect the expression of XIST in cisplatin-resistant human nasopharyngeal carcinoma cell line HNE1/DDP. The effects of up-regulation and down-regulation of XIST on DDP resistance, proliferation and apoptosis of HNE1/ DDP cells were assessed using MTT assay, EdU assay and flow cytometry. Western blotting was used to detect the changes in the expressions of programmed cell death 4 (PDCD4) and Fas ligand (Fas-L) proteins in the cells in response to up-regulation or down-regulation of XIST. Results The expression of XIST was significantly up-regulated in HNE1/DDP cells in comparison with HNE1 cells (0.57±0.06 vs 0.1±0.02, P<0.05). Down-regulation of XIST significantly decreased while up-regulation of XIST obviously increased DDP resistance of HNE1/DDP cells (P<0.05). Down-regulation of XIST significantly reduced the proliferation (6.17 ± 1.93 vs 16.59 ± 4.86, P<0.05) and enhanced apoptosis [(18.04 ± 4.72)% vs (4.22 ± 1.65)% , P<0.05], while upregulating XIST enhanced the proliferation (25.40±7.21 vs 13.16±3.95, P<0.05) and inhibited apoptosis [(2.82±0.88)% vs (6.46± 1.75)%, P<0.05] in HNE1/DDP cells. Down-regulation of XIST significantly increased the protein expressions of PDCD4 and Fas-L (P<0.05) in HNE1/DDP cells, and up-regulation of XIST resulted in reverse changes in PDCD4 and Fas-L expressions (P< 0.05). Conclusion XIST is up-regulated in HNE1/DDP cells, and down-regulation and up-regulation of XIST expression reduce and increase DDP resistance of the cells, respectively, possibly as a result of changes in the expressions of PDCD4 and Fas-L.