基于食管鳞癌奥沙利铂耐药细胞测序差异表达分析  

Analysis of differential expression based on sequencing of oxaliplatin resistant cells in esophageal squamous cancer

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作  者:董慧玲 刘祥辰 孔鹏洲[3] 徐恩伟[4] 宋彬 DONG Hui-ling;LIU Xiang-chen;KONG Peng-zhou;XU En-wei;SONG Bin(不详;The First Hospital of Shanxi Medical University,Taiyuan 030001,Shanxi Province,China)

机构地区:[1]山西医科大学第一医院,山西太原030001 [2]中山大学附属第五医院,广东珠海519000 [3]山西医科大学医学转化中心,山西省肿瘤医院,山西太原030001 [4]山西医科大学白求恩医院,山西太原030001

出  处:《中国生物制品学杂志》2022年第12期1443-1448,1457,共7页Chinese Journal of Biologicals

基  金:国家自然科学基金(81703016);山西省自然科学基金(20180D121306);山西白求恩医院2020年度院级肿瘤专项公开基金(2020-ZL02)。

摘  要:目的建立食管鳞癌(esophageal squamous cancer,ESCC)奥沙利铂(oxaliplatin,L-OHP)耐药细胞株,全转录本测序筛选与L-OHP耐药相关的基因,探索L-OHP耐药机制。方法采用L-OHP大剂量冲击与逐步递增药物浓度的方法建立人ESCC L-OHP耐药细胞株KYSE150/L-OHP,采用CCK-8法鉴定耐药细胞株的耐药性;光学显微镜下观察耐药细胞的形态学改变;Transwell小室试验检测耐药细胞侵袭/迁移能力的变化;全转录本测序筛选KYSE150和KYSE150/L-OHP细胞差异表达基因,并对差异表达基因进行KEGG通路富集和RT-PCR验证。结果L-OHP耐药细胞模型KYSE150/L-OHP的耐药指数为(4.0±1.35),明显高于亲本细胞(t=22.9,P<0.01);光镜下KYSE150/L-OHP细胞体积增大,形态不规则;与亲本细胞株相比,KYSE150/L-OHP细胞的侵袭和迁移能力均显著增强(t分别为25.49和46.05,P均<0.0001);全转录本测序结果显示,P13K-Akt、MAPK、Hippo信号通路可能介导L-OHP耐药的发生过程;RT-PCR验证差异基因表达与测序结果一致。结论成功建立了人ESCC L-OHP耐药细胞模型KYSE150/L-OHP,全转录本测序技术探讨了L-OHP化疗耐药的分子机制,为临床晚期ESCC L-OHP耐药患者的治疗提供了思路。Objective To develop oxaliplatin(L-OHP)resistant cell lines for esophageal squamous cancer(ESCC),perform complete transcript sequencing to screen genes related to L-OHP resistance,and explore the mechanism of it.Methods The human ESCC L-OHP resistant cell line KYSE150/L-OHP was developed by high dose shock of L-OHP and gradual increase in drug concentration,which was identified for drug resistance by CCK-8,observed for morphological changes under optical microscope and determined for change of invasion or migration ability by Transwell assay.The differentially expressed genes of KYSE150 and KYSE150/L-OHP cells were screened by complete transcript sequencing,which were enriched by KEGG pathway and verified by RT-PCR.Results The drug resistance index of L-OHP resistant cell model KYSE150/L-OHP was(4.0±1.35),which was significantly higher than that of parent cells(t=22.9,P<0.01).KYSE150/L-OHP cells showed increased volume and irregular morphology under optical microscope.Compared with that of parent cell lines,the invasion and migration ability of KYSE150/L-OHP cells were both significantly enhanced(t=25.49 and 46.05 respectively,each P<0.0001).Complete transcript sequencing showed that P13K-Akt,MAPK and Hippo signaling pathways may mediate the development of L-OHP resistance,which was consistent with the differential gene expressions verified by RT-PCR.Conclusion A human ESCC L-OHP resistant cell model,KYSE150/L-OHP,was successfully developed and the molecular mechanism of L-OHP chemotherapy resistance was explored by complete transcript sequencing technology,which provided ideas for the treatment of patients with advanced clinical ESCC L-OHP resistance.

关 键 词:食管麟癌 奥沙利铂 耐药 全转录本测序 通路分析 

分 类 号:R73-3[医药卫生—肿瘤]

 

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