Li Haiyan, Wang Jinwei, Jin Yingying, Lin Jian, Gong Liuyang, Xu Youzu
Department of Respiratory and Critical Care Medicine, Taizhou Hospital of Zhejiang Province Affiliated to Wenzhou Medical University, Linhai, China.
Functional Inspection Section, Taizhou Hospital of Zhejiang Province affiliated to Wenzhou Medical University, Linhai, China.
Transl Cancer Res. 2022 Aug;11(8):2876-2886. doi: 10.21037/tcr-22-1812.
The incidence of lung cancer is extremely high. For treatment, the chemotherapy of lung cancer is low, and drug resistance and recurrence are frequently observed, in which hypoxic tumor microenvironments play a key role. We investigated the upregulation of long non-coding ribonucleic acid (lncRNA) H19 expression and the induction of drug resistance in non-small cell lung cancer (NSCLC) cells under hypoxic conditions.
We used cobaltous chloride (CoCl2) to create hypoxic culture environments for the A549 and H1650 cells, and they were divided into normoxia and hypoxia groups. We used real-time quantitative PCR detecting system (qPCR) to detect the messenger RNA (mRNA) expressions of lncRNA H19, hypoxia-inducible factor 1-α (HIF1-α), multi-drug resistant associated protein 1 (MRP1), and heme oxygenase-1 (HO-1), and western blot to detect the protein expressions of HIF1-α, MRP1, HO-1, and P-glycoprotein (P-gp). We performed cell migration and invasion assays. Annexin V-Allophycocyanin (APC) and flow cytometry were used to detect the apoptotic rates.
We found that the expression levels of lncRNA H19, HIF1-α, HO-1, multi-drug resistant associated protein 1 (MRP1), and P-glycoprotein increased significantly in the NSCLC cell lines (A549 and H1650) under hypoxic conditions, as determined by the qPCR and western blot analyses. In NSCLC cells cultured under hypoxic conditions, the invasion and migration ability of tumor cells increased significantly, resistance to cisplatin increased, and cisplatin-induced apoptosis decreased considerably. In addition, chemoresistance was also induced in tumor cells under hypoxic conditions.
These results indicate that hypoxia increased the expression levels of lncRNA H19 and induced chemoresistance in tumor cells.
肺癌发病率极高。在治疗方面，肺癌化疗效果欠佳，且常出现耐药和复发情况，其中缺氧肿瘤微环境起关键作用。我们研究了缺氧条件下非小细胞肺癌（NSCLC）细胞中长链非编码核糖核酸（lncRNA）H19表达上调及耐药性诱导情况。
我们用氯化钴（CoCl2）为A549和H1650细胞创建缺氧培养环境，并将其分为常氧组和缺氧组。我们使用实时定量PCR检测系统（qPCR）检测lncRNA H19、缺氧诱导因子1-α（HIF1-α）、多药耐药相关蛋白1（MRP1）和血红素加氧酶-1（HO-1）的信使核糖核酸（mRNA）表达，并用蛋白质印迹法检测HIF1-α、MRP1、HO-1和P-糖蛋白（P-gp）的蛋白质表达。我们进行了细胞迁移和侵袭试验。使用膜联蛋白V-别藻蓝蛋白（APC）和流式细胞术检测凋亡率。
通过qPCR和蛋白质印迹分析确定，我们发现缺氧条件下NSCLC细胞系（A549和H1650）中lncRNA H19、HIF1-α、HO-1、多药耐药相关蛋白1（MRP1）和P-糖蛋白的表达水平显著增加。在缺氧条件下培养的NSCLC细胞中，肿瘤细胞的侵袭和迁移能力显著增强，对顺铂的耐药性增加，顺铂诱导的凋亡显著减少。此外，缺氧条件下肿瘤细胞也产生了化学抗性。
这些结果表明，缺氧增加了lncRNA H19的表达水平并诱导肿瘤细胞产生化学抗性。
