LncRNA HOTAIR通过靶向miR-126激活SHH信号通路促进结直肠癌的发生发展

LncRNA HOTAIR activating SHH signaling pathway by targeting miR-126 to promote the occurrence and development of colorectal cancer

目的:探究miR-126与lncRNA HOTAIR的靶向关系及其对SHH信号通路的调节作用,并研究结直肠癌发生发展的分子机制。方法:收集2018年2月至2019年2月于河北省沧州市人民医院进行手术治疗的患者的正常结直肠、结直肠息肉、不典型增生组织、癌前病变、结直肠癌组织及其癌旁正常组织标本,采用实时荧光定量聚合酶链式反应(RT-qPCR)、免疫组化分析HOTAIR、miR-126及SHH表达情况,starBase数据库、荧光素酶实验、RNA免疫共沉淀及RT-qPCR验证miR-126与HOTAIR、SHH的作用关系,MTT实验、集落形成实验、划痕实验、Transwell实验检测细胞活力、增殖、迁移、侵袭能力。结果:结直肠癌组织中的HOTAIR和SHH、Glil、Smo蛋白表达上调,miR-126表达下调(P<0.05)。HOTAIR靶向抑制miR-126表达,miR-126靶向抑制SHH表达。与sh-NC组相比,sh-HOTAIR组SW480、HCT116细胞活力、增殖、迁移、侵袭能力下降,miR-126 inhibitor组增加(P<0.05);sh-HOTAIR+miR-126 inhibitor组细胞的活力、增殖、迁移、侵袭能力显著高于sh-HOTAIR组,低于miR-126 inhibitor组(P<0.05);miR-126 inhibitor+sh-SHH组细胞的活力、增殖、迁移、侵袭能力显著低于miR-126 inhibitor组(P<0.05)。结论:HOTAIR靶向抑制miR-126表达,从而促进结直肠癌的发生发展,其机制可能与激活SHH信号通路的表达有关。

Objective:To investigate the targeting relationship between miR-126 and lncRNA HOTAIR and its regulatory effect on SHH signaling pathway,and to study the molecular mechanism in the occurrence and development of colorectal cancer.Methods:Samples of normal colorectal tissues,colorectal polyps,atypical hyperplasia,precancerous lesions,colorectal cancer tissues and adjacent normal tissues were obtained from patients who underwent surgery in Cangzhou People’s Hospital of Hebei Province from February 2018 to February 2019.Real-time fluorescence quantitative polymerase chain reaction(RT-qPCR)and immunohistochemistry were used to analyze the expression of HOTAIR,miR-126 and SHH.StarBase,luciferase assay,RNA co-immunoprecipitation and RT-qPCR were used to verify the relationship of miR-126 with HOTAIR and SHH.MTT,colony-forming assay,scratch assay and Transwell test were performed to detect cell viability,proliferation,migration and invasion.Results:The expression of HOTAIR and SHH,Glil,Smo proteins in colorectal cancer tissues were significantly up-regulated,and the expression of miR-126 was significantly down-regulated(P<0.05).HOTAIR targeted to inhibit the expression of miR-126,and miR-126 targeted to inhibit the expression of SHH.Compared with the sh-NC group,the cell viability,proliferation,migration and invasion of SW480 and HCT116 cells in sh-HOTAIR group decreased,while those in the miR-126 inhibitor group increased(P<0.05).The cell viability,proliferation,migration and invasion of cells in the sh-HOTAIR+miR-126 inhibitor group were significantly higher than those in the sh-HOTAIR group,and significantly lower than those in the miR-126 inhibitor group (P<0.05). The cell viability, proliferation, migration and invasionof cells in the miR-126 inhibitor+sh-SHH group were significantly lower than those in the miR-126 inhibitorgroup (P<0.05). Conclusion: The expression of HOTAIR targets to inhibit the expression of miR-126, therebypromoting the occurrence and development of colorectal cancer. The mechanism may be related to the activationof SHH signaling pathway.