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目的:探讨miR-128对非小细胞肺癌发生发展及血管和淋巴管生成的影响及相关机制。方法:RT-PCR检测肿瘤组织及细胞miR-128的表达;应用慢病毒表达载体建立稳定表达miR-128的非小细胞肺癌细胞株;裸鼠移植瘤治疗实验证实ERK,AKT和p38信号通路的磷酸化活性降低,进而阻断了上述信号通路。结果:miR-128在肺癌细胞株和非小细胞肺癌组织中表达下调,并且与非小细胞肺癌的分化程度、病理分期和淋巴结转移相关。异常的miR-128高表达可明显抑制肺癌细胞株的体外增殖、克隆形成、迁移和侵袭能力,并且诱使细胞发生G1期细胞阻滞和凋亡发生。更重要的是,荧光素酶系统证实了miR-128与VEGF-C的靶向关系,miR-128的高表达促使了肿瘤细胞VEGF-C的表达下调和VEGF-C 3’UTR荧光素酶活性降低,导致了调控肿瘤血管和淋巴管生成的VEGF家族因子VEGF-A,VEGFR-2和VEGFR-3的表达下调,使得ERK,AKT和p38信号通路的磷酸化活性降低,进而阻断了上述信号通路。而且裸鼠移植瘤治疗实验证实,体内恢复miR-128的高表达,可抑制肿瘤的体内生长及血管和淋巴管的生成。结论:miR-128在非小细胞肺癌的发生中起着重要作用,至少可以通过靶向调控VEGF-C表达来同时控制肿瘤的血管和淋巴管生成及相关信号通路,体内恢复miR-128的高表达有望成为非小细胞肺癌治疗的新策略。
Objective: To investigate the effects of miR-128 on the development and progression of non-small cell lung carcinoma (NSCLC) and angiogenesis and lymphangiogenesis and related mechanisms. Methods: The expression of miR-128 in tumor tissues and cells was detected by RT-PCR. The lentiviral expression vector was used to establish non-small cell lung cancer cell line stably expressing miR-128. The transplanted tumor in nude mice proved that ERK, AKT and p38 signaling pathways Phosphorylation activity decreased, thereby blocking the signal pathway. Results: miR-128 was down-regulated in lung cancer cell lines and non-small cell lung cancer tissues, and correlated with the degree of differentiation, pathological stage and lymph node metastasis of non-small cell lung cancer. Aberrant miR-128 overexpression significantly inhibited the proliferation, colony formation, migration and invasion of lung cancer cell lines in vitro and induced cell arrest and apoptosis in G1 phase. More importantly, the luciferase system confirmed the targeted relationship between miR-128 and VEGF-C. The high miR-128 expression led to the down-regulation of VEGF-C expression and VEGF-C 3’UTR luciferase activity in tumor cells Decreased the expression of VEGF family factors VEGF-A, VEGFR-2 and VEGFR-3 that regulate tumor angiogenesis and lymphangiogenesis, decreased the phosphorylation activity of ERK, AKT and p38 signaling pathway, and then blocked the above signal path. In addition, the treatment of transplanted xenografts in nude mice confirmed that the high expression of miR-128 in vivo can inhibit the growth of tumors and the formation of blood vessels and lymphatic vessels in vivo. Conclusions: miR-128 plays an important role in the development of non-small cell lung cancer. At least miR-128 can be restored in vivo by at least regulating the expression of VEGF-C and simultaneously controlling tumor angiogenesis, lymphangiogenesis and related signaling pathways Expression is expected to become a new strategy for the treatment of non-small cell lung cancer.