miR-362-3p的表达及其靶基因与胆囊癌恶性特征的关系

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目的:探讨miR-362-3p在胆囊癌中的表达及功能.方法:用qRT-PCR检测手术切除的44例胆囊癌患者手术标本中miR-362-3P的表达,并分析其表达与胆囊癌临床病理特征及预后的关系.miR-363-3p模拟物转染胆囊癌细胞后,分别用MTT法、细胞划痕试验及Transwell侵袭试验观察细胞增殖、迁移及侵袭的改变.通过生物信息学方法及双荧光素酶报告基因试验分析miR-362-3p的靶基因,并采用补救试验验证.结果:miR-362-3p在胆囊癌组织的表达明显低于相应癌旁组织(P<0.05).miR-362-3p的低表达与肿瘤TNM分期、淋巴结转移及远处转移明显有关(均P<0.05).低表达miR-362-3p患者总体生存率较高表达miR-362-3p患者明显降低(P<0.05).转染miR-362-3p模拟物后,胆囊癌细胞的增殖,迁移及侵袭能力明显减弱(均P<0.05).Nemo样激酶(NLK)被确定为miR-362-3p的潜在靶基因,转染 NLK过表达载体后,miR-362-3p模拟物对胆囊癌细胞的上述作用被明显逆转(均P<0.05 ).结论:miR-362-3p在胆囊癌中表达下调,下调的miR-362-3p减少了对靶基因NLK的抑制,从而促进了胆囊癌细胞的增殖、迁移和侵袭.“,”Objective: To detect the expression of miR-362-3p in gallbladder cancer and its functions. Methods: The expressions of miR-362-3p in the surgical specimens from 44 gallbladder cancer patients were determined by qRT-PCR, and the relations of miR-362-3p expression with clinicopathological characteristics and prognosis of the patients were analyzed. In gallbladder cancer cells after transfection with miR-362-3p mimics, the changes in proliferation, migration and invasion were examined by MTT colorimetry, wound healing assay, and Transwell assay respectively. The target gene of miR-362-3p was analyzed by bioinformatics analysis and dual luciferase reporter gene assay, and then was verified through recovery tests. Results: The expression of miR-362-3p was significantly decreased in gallbladder cancer tissue compared with tumor adjacent tissue (P<0.05). The decreased miR-362-3p expression was significantly associated with tumor TNM stage, lymph node metastasis and distant metastasis (all P<0.05). The overall survival rate in patients with low miR-362-3p expression was significantly lower than that in patients with high miR-362-3p expression (P<0.05). The abilities of proliferation, migration and invasion were significantly reduced in gallbladder cancer cells after transfection with miR-362-3p mimics (all P<0.05). Nemo like kinase (NLK) was found potentially to be the target gene of miR-362-3p. After transfection with NLK overexpression vectors,the above effects exerted by miR-362-3p mimics in gallbladder cancer cells were significantly reversed (all P<0.05). Conclusion: The expression of miR-362-3p is down-regulated in gallbladder cancer, and the down-regulated miR-362-3p expression may reduce the inhibition on its target gene NLK, and thereby promote the proliferation, migration and invasion of gallbladder cancer cells.
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