Cai Yuanming, Jia Ruxue, Xiong Haozhe, Ren Qun, Zuo Weimin, Lin Tingting, Lin Rong, Lei Yan, Wang Ping, Dong Huiyue, Zhao Hu, Zhu Ling, Fu Yunfeng, Zeng Zhiyong, Zhang Wei, Wang Shuiliang
Undergraduate Grade 2015, The 5th Clinical Medical School of Xinjiang Medical University, Urumchi, People's Republic of China.
Department of Urology, The 900th Hospital of the Joint Logistics Team, Fujian Medical University, Fuzhou, Fujian, People's Republic of China.
Cancer Manag Res. 2019 Aug 5;11:7391-7404. doi: 10.2147/CMAR.S215427. eCollection 2019.
Paclitaxel has shown significant anti-tumor activity against non-small cell lung cancer (NSCLC); however, resistance to paclitaxel frequently occurs and represents a significant clinical problem and its underlying molecular mechanism remains elusive.
Long-term treatment of culture cell with paclitaxel was carried out to mimic the development of acquired drug resistance in NSCLC. Cell proliferation and clonogenic assay and apoptosis evaluation were carried out to determine the efficacy of paclitaxel on NSCLC cells. Western blot analyses were performed to determine the expression and activation of proteins. Apoptosis enzyme-linked immunosorbent assay was used to quantify cytoplasmic histone-associated DNA fragments. Microarray analyses were applied to explore both mRNA and miRNA expression profiles in NSCLC cells followed by integrative analysis. qRT-PCR was carried out to verify the differentially expressed mRNAs and miRNAs.
The expression of 652 genes was shown to be changed at least 2-fold in paclitaxel-resistant NSCLC (H460_TaxR) cells with 511 upregulated and 141 downregulated as compared with that in parental H460 cells. The differentially expressed genes were functionally enriched in regulating the cell proliferation, cell death, and response to endogenous stimulus, and clustered in pathways such as cancer and signaling by the G protein-coupled receptor (GPCR). Moreover, 43 miRNAs were shown to be differentially expressed in H460_TaxR cells with 15 upregulated and 28 downregulated as compared with parental H460 cells. A total of 289 pairs of miRNA-potential target gene were revealed in H460_TaxR cells by bioinformatics analysis. Furthermore, integrative analysis of miRNAs and gene expression profiles revealed that dysregulated miR-362-3p, miR-766-3p, and miR-6507-3p might confer paclitaxel resistance in NSCLC via targeting MAPT simultaneously.
Our findings suggested that specific manipulation of MAPT-targeting miRNAs may be a novel strategy to overcome paclitaxel resistance in patients with NSCLC especially large-cell lung carcinoma.
紫杉醇已显示出对非小细胞肺癌（NSCLC）具有显著的抗肿瘤活性；然而，对紫杉醇的耐药性经常出现，这是一个重大的临床问题，其潜在的分子机制仍然不清楚。
用紫杉醇对培养细胞进行长期处理，以模拟NSCLC中获得性耐药的发展过程。进行细胞增殖、克隆形成试验和凋亡评估，以确定紫杉醇对NSCLC细胞的疗效。进行蛋白质印迹分析以确定蛋白质的表达和激活情况。采用凋亡酶联免疫吸附测定法对细胞质中组蛋白相关DNA片段进行定量。应用微阵列分析来探索NSCLC细胞中的mRNA和miRNA表达谱，随后进行综合分析。进行qRT-PCR以验证差异表达的mRNA和miRNA。
与亲本H460细胞相比，在耐紫杉醇的NSCLC（H460_TaxR）细胞中，有652个基因的表达变化至少达2倍，其中511个上调，141个下调。差异表达的基因在调节细胞增殖、细胞死亡和对内源性刺激的反应方面功能富集，并聚集在癌症和G蛋白偶联受体（GPCR）信号传导等途径中。此外，与亲本H460细胞相比，在H460_TaxR细胞中有43个miRNA差异表达，其中15个上调，28个下调。通过生物信息学分析在H460_TaxR细胞中总共揭示了289对miRNA-潜在靶基因。此外，对miRNA和基因表达谱的综合分析表明，失调的miR-362-3p、miR-766-3p和miR-6507-3p可能通过同时靶向MAPT赋予NSCLC对紫杉醇的耐药性。
我们的研究结果表明，特异性调控靶向MAPT的miRNA可能是克服NSCLC患者尤其是大细胞肺癌患者对紫杉醇耐药的一种新策略。
