Department of Pathology, Shandong Provincial Hospital Affiliated to Shandong University, Jinan, China.
Eur Rev Med Pharmacol Sci. 2019 Apr;23(7):2827-2837. doi: 10.26355/eurrev_201904_17559.
This study was to find out the influence of microRNA-129-5p on proliferative ability, invasiveness, and metastasis of lung tumor cells and tumor angiogenesis. Besides, the effects of microRNA-129-5p on vascular endothelial growth factor (VEGF) level and potential regulatory mechanisms were also what we were interested in.
Quantitative Real Time-Polymerase Chain Reaction (qRT-PCR) assay was performed to detect the microRNA-129-5p level in tumor tissues and paracancerous tissues of 50 patients with LCa, and the interaction between microRNA-129-5p expression and LCa pathological parameters was analyzed. The untreated cell group (NC) and the transfected microRNA-129-5p overexpression group (microRNA-129-5p mimics) were established, and then, the transfection efficiency of microRNA-129-5p was further verified by qRT-PCR. In H1299 and SPC-A1, cell counting kit-8 (CCK-8), Tube-formation experiments, and transwell invasion and migration tests were performed to evaluate the influence of the microRNA on the biological function of LCa cells. Finally, the potential mechanism of action of VEGF, a downstream gene of microRNA-129-5p, was explored by bioinformatics analysis and recovery experiments.
QRT-PCR results showed that the level of microRNA-129-5p in cancer tissues of LCa patients was notably lower than that in normal tissues, and the difference was statistically significant. Compared with patients with highly expressed microRNA-129-5p, patients with low level had higher rates of lymph node or distant metastasis, and the overall survival rate was lower. Compared with NC group, cell proliferation, invasiveness and migration ability, and tumor angiogenesis capacity were strikingly decreased in microRNA-129-5p mimics group. Subsequently, VEGF expression was validated conspicuously enhanced in LCa cell line and tissue and was negatively correlated with microRNA-129-5p. In addition, the recovery experiment demonstrated that overexpression of VEGF could counteract the impact of microRNA-129-5p mimics on tumor angiogenesis and the invasive and migratory capacity of LCa cells, which then together led to the malignant progression of LCa.
The above studies demonstrated that microRNA-129-5p was strikingly correlated with LCa lymph node or distant metastasis and poor prognosis, and it can inhibit the malignant progression of this cancer. The investigation also demonstrated that microRNA-129-5p may inhibit proliferation capacity and invasiveness of LCa cells and tumor angiogenesis via regulating VEGF.
本研究旨在探讨 microRNA-129-5p 对肺肿瘤细胞增殖能力、侵袭性和转移以及肿瘤血管生成的影响。此外,我们还关注 microRNA-129-5p 对血管内皮生长因子 (VEGF) 水平的影响及其潜在的调节机制。
采用实时定量聚合酶链反应 (qRT-PCR) 检测 50 例 LCa 患者肿瘤组织和癌旁组织中的 microRNA-129-5p 水平,并分析 microRNA-129-5p 表达与 LCa 病理参数之间的相互关系。建立未处理细胞组 (NC) 和转染 microRNA-129-5p 过表达组 (microRNA-129-5p 模拟物),进一步通过 qRT-PCR 验证 microRNA-129-5p 的转染效率。在 H1299 和 SPC-A1 中,通过细胞计数试剂盒-8 (CCK-8)、管形成实验、Transwell 侵袭和迁移实验评估 microRNA 对 LCa 细胞生物学功能的影响。最后,通过生物信息学分析和恢复实验探讨 VEGF(microRNA-129-5p 的下游基因)的潜在作用机制。
qRT-PCR 结果显示,与正常组织相比,LCa 患者癌组织中 microRNA-129-5p 水平明显降低,差异具有统计学意义。与高表达 microRNA-129-5p 的患者相比,低水平 microRNA-129-5p 的患者淋巴结或远处转移率更高,总生存率更低。与 NC 组相比,microRNA-129-5p 模拟物组细胞增殖、侵袭和迁移能力以及肿瘤血管生成能力明显降低。随后,在 LCa 细胞系和组织中验证了 VEGF 表达明显增强,且与 microRNA-129-5p 呈负相关。此外,恢复实验表明,过表达 VEGF 可拮抗 microRNA-129-5p 模拟物对 LCa 细胞血管生成和侵袭迁移能力的影响,进而共同导致 LCa 的恶性进展。
上述研究表明,microRNA-129-5p 与 LCa 淋巴结或远处转移及预后不良显著相关,可抑制该癌症的恶性进展。研究还表明,microRNA-129-5p 可能通过调节 VEGF 抑制 LCa 细胞增殖能力、侵袭性和肿瘤血管生成。