Department of Surgery, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi 530021, P.R. China.
Department of Anaesthesiology, The First Affiliated Hospital of Guangxi Medical University, Nanning, Guangxi 530021, P.R. China.
Int J Mol Med. 2018 Apr;41(4):1958-1966. doi: 10.3892/ijmm.2018.3381. Epub 2018 Jan 11.
Due to a lack of effective methods for early diagnosis, the majority of patients with gastric cancer (GC) are diagnosed during the late stages of the disease, which are often accompanied by metastasis. For these patients, despite being considered an important therapeutic modality in the treatment of cancer, chemotherapy is usually not effective due to multidrug resistance (MDR). The expression levels of MDR/metastasis‑associated genes are regulated by numerous microRNAs (miRNAs/miRs). The expression of miR-647 in GC tissues and SGC7901/VCR cell line (drug resistance to vincristine) was detected by qRT-PCR. The effect of overexpression of miR-647 on drug resistance was evaluated by measuring the half maximal inhibitory concentration (IC50) value of SGC-7901/VCR to vincristine and tumor growth in vivo. Moreover, drug-induced cell apoptosis and cell cycle were evaluated by flow cytometry, as well as the ability of cell migration and invasiveness detected by wound healing and transwell assay. Furthermore, underlying targets of miR-647 were predicted by TargetScan and MicroRNA; meanwhile, the expression of ANK2, FAK, MMP2, MMP12,CD44,SNAIL1 were observed by qRT-PCR and western blot analysis. The present study established that the expression levels of miR‑647 were downregulated in GC tissues from patients with metastasis and in the vincristine‑resistant SGC7901 (SGC‑7901/VCR) GC cell line. The IC50 value for vincristine was significantly decreased, whereas the proportion of cells in G0/G1 phase and the drug‑induced apoptotic rate were significantly increased following upregulation of miR‑647. Furthermore, the results demonstrated that miR‑647 overexpression led to decreased migration and invasion of SGC‑7901/VCR cells. Overexpression of miR‑647 was also demonstrated to sensitize tumors to chemotherapy in vivo. In addition, miR‑647 overexpression was able to reduce the expression levels of ankyrin‑B, focal adhesion kinase, matrix metalloproteinase (MMP)2, MMP12, cluster of differentiation 44 and snail family transcriptional repressor 1. In conclusion, these findings demonstrated that miR‑647 may function as a novel target to ameliorate drug resistance and metastasis of GC cells.
由于缺乏早期诊断的有效方法，大多数胃癌（GC）患者在疾病晚期被诊断出来，此时通常伴有转移。对于这些患者，尽管化疗被认为是癌症治疗的重要治疗方式，但由于多药耐药（MDR），通常效果不佳。MDR/转移相关基因的表达水平受许多 microRNAs（miRNAs/miRs）的调节。通过 qRT-PCR 检测 GC 组织和 SGC7901/VCR 细胞系（对长春新碱耐药）中 miR-647 的表达。通过测量 SGC-7901/VCR 对长春新碱的半最大抑制浓度（IC50）值和体内肿瘤生长来评估 miR-647 过表达对耐药性的影响。此外，通过流式细胞术评估药物诱导的细胞凋亡和细胞周期，通过划痕愈合和 Transwell 测定检测细胞迁移和侵袭能力。此外，通过 TargetScan 和 MicroRNA 预测 miR-647 的潜在靶标；同时，通过 qRT-PCR 和 Western blot 分析观察 ANK2、FAK、MMP2、MMP12、CD44、SNAIL1 的表达。本研究表明，miR-647 的表达水平在有转移的 GC 组织和长春新碱耐药的 SGC7901（SGC-7901/VCR）GC 细胞系中下调。长春新碱的 IC50 值显著降低，而 G0/G1 期细胞比例和药物诱导的凋亡率显著增加，miR-647 上调。此外，结果表明 miR-647 过表达导致 SGC-7901/VCR 细胞迁移和侵袭减少。体内过表达 miR-647 也能使肿瘤对化疗敏感。此外，miR-647 过表达能够降低锚蛋白-B、粘着斑激酶、基质金属蛋白酶（MMP）2、MMP12、分化群 44 和蜗牛家族转录阻遏物 1 的表达水平。总之，这些发现表明 miR-647 可能作为一种新的靶点，改善 GC 细胞的耐药性和转移。
