Department of Laboratory Medicine, Key Laboratory of Clinical Laboratory Diagnosis and Translation Research of Zhejiang Province, The First Affiliated Hospital of Wenzhou Medical University, Nanbaixiang Street, Ouhai District, Wenzhou, 325000, Zhejiang Province, China.
J Cancer Res Clin Oncol. 2023 Mar;149(3):1295-1311. doi: 10.1007/s00432-022-04152-5. Epub 2022 Jul 6.
Our previous experiments have demonstrated that lncRNA UCA1 (UCA1) promoted cisplatin resistance in lung adenocarcinoma (LUAD). This study aimed to explore the potential downstream target genes regulated by UCA1 and how this downstream gene promotes cisplatin resistance in LUAD.
Here, we measured the expression level of Heme oxygenase1 (HO1) in LUAD cell lines by reverse transcription-quantitative polymerase chain reaction (RT-qPCR) based on UCA1 overexpression cell lines and UCA1 knockdown cell lines. HO1 was knocked down in the UCA1 overexpression cell line, and HO1 was overexpressed in the UCA1 knockdown cell line, and the half maximal inhibitory concentration (IC50) trends were observed by adding cisplatin containing a certain concentration gradient. Cell functional assays were performed to observe the changes in the biological behavior of HO1 after overexpression and knockdown, and the tumorigenic assay in nude mice was performed to verify the effect of UCA1 in regulating the growth and cisplatin resistance of HO1 on LUAD cells in vivo.
The results showed that HO1 and UCA1 expression were both upregulated in LUAD tissues and LUAD cisplatin-resistant cell lines, and there was a significant positive correlation between the expression of HO1 and UCA1. In vitro experiments showed that HO1 overexpression could reverse the reduced sensitivity to cisplatin caused by UCA1 knockdown in A549/DDP cells, and HO1 knockdown could reduce cisplatin resistance in A549 UCA1 overexpressing cells. Tumorigenic assays in nude mice further confirmed the role of HO1 in the regulation of UCA1 by activating the NRF2/HO1 pathway against LUAD cisplatin resistance.
Our findings suggested that UCA1 regulates HO1 targets the UCA1/NRF2-HO1 pathway to exert cisplatin resistance in LUAD.
我们之前的实验已经证明长链非编码 RNA UCA1（UCA1）促进了肺腺癌（LUAD）对顺铂的耐药性。本研究旨在探索 UCA1 调控的潜在下游靶基因，以及该下游基因如何促进 LUAD 对顺铂的耐药性。
在这里，我们通过基于 UCA1 过表达细胞系和 UCA1 敲低细胞系的逆转录定量聚合酶链反应（RT-qPCR）测量 LUAD 细胞系中血红素加氧酶 1（HO1）的表达水平。在 UCA1 过表达细胞系中敲低 HO1，并在 UCA1 敲低细胞系中过表达 HO1，观察添加含有一定浓度梯度的顺铂后 IC50 趋势。进行细胞功能测定以观察 HO1 过表达和敲低后生物学行为的变化，并在裸鼠中进行肿瘤发生测定以验证 UCA1 调节 HO1 在体内对 LUAD 细胞生长和顺铂耐药性的作用。
结果表明，HO1 和 UCA1 的表达在 LUAD 组织和 LUAD 顺铂耐药细胞系中均上调，并且 HO1 和 UCA1 的表达之间存在显著的正相关。体外实验表明，在 A549/DDP 细胞中，HO1 过表达可以逆转 UCA1 敲低导致的对顺铂敏感性降低，而在 A549 UCA1 过表达细胞中，HO1 敲低可以降低顺铂耐药性。裸鼠肿瘤发生实验进一步证实了 HO1 在 UCA1 通过激活 NRF2/HO1 通路调节对 LUAD 顺铂耐药性中的作用。
我们的研究结果表明，UCA1 通过调控 UCA1/NRF2-HO1 通路来调节 HO1 靶基因，从而在 LUAD 中发挥顺铂耐药性。
