miR-671-5p通过靶向肿瘤坏死因子α诱导蛋白8调控胰腺癌细胞增殖和凋亡

目的：探讨miR-671-5p 靶向肿瘤坏死因子α 诱导蛋白8（ TNFAIP8）对胰腺癌细胞增殖和凋亡的影响。 方法：实时荧光定量PCR（ qRT-PCR）和免疫印迹检测20 例胰腺癌组织和与其配对的癌旁正常组织miR-617-5p、 TNFAIP8 mRNA和TNFAIP8表达水平，验证miR-671-5p 和TNFAIP8的靶向调控关系。将体外培养胰腺癌细胞 capan-1分为miR-NC组、miR-671-5p 组、si-NC 组、si-TNFAIP8 组、miR-671-5p+pcDNA组和miR-671-5p+pcDNATNFAIP8 组。采用四甲基偶氮唑蓝（ MTT）实验检测细胞活力，流式细胞术检测细胞凋亡，免疫印迹检测细胞周 期蛋白D1（ cyclin D1）、p21、B细胞淋巴瘤/ 白血病-2（ Bcl-2）和Bcl-2 相关蛋白（ Bax）的表达水平。结果：与 癌旁正常组织比较，胰腺癌组织miR-617-5p 表达量显著降低，TNFAIP8 的表达量显著升高。miR-671-5p 靶向负向 调控TNFAIP8 表达。与miR-NC组比较，miR-671-5p 组capan-1 细胞活力显著降低，细胞凋亡率显著升高，cyclin D1和Bcl-2 的表达量显著降低，p21 和Bax 的表达量显著升高；与si-NC 组比较，si-TNFAIP8 组capan-1 细胞活力 显著降低，细胞凋亡率显著升高，cyclin D1和Bcl-2 表达量显著降低，p21 和Bax 表达量显著升高；与miR-671- 5p+pcDNA 组比较，miR-671-5p+pcDNA-TNFAIP8 组capan-1 细胞活力显著升高，细胞凋亡率显著降低，cyclin D1和Bcl-2 的表达量显著升高，p21 和Bax 的表达量显著降低。结论：miR-671-5p 通过靶向下调TNFAIP8 抑制胰 腺癌细胞增殖并促进其凋亡。上调miR-671-5p 是胰腺癌潜在治疗靶点。

miR-671-5p regulates the proliferation and apoptosis of pancreatic cancer cells by targeting TNFAIP8

Objective To investigate the effect of miR-671-5p on proliferation and apoptosis of pancreatic cancer cells by targeting tumour necrosis factor-alpha-induced protein 8（ TNFAIP8）. Methods Real-time quantitative PCR （ qRT-PCR） and Western blotting were used to detect the expression levels of miR-617-5p， TNFAIP8 mRNA and TNFAIP8 protein in 20 pancreatic cancer tissues and their adjacent paracancerous tissues. The dual luciferase reporter gene assay and Western blotting were used to verify the targeted and regulatory relationship between miR-671-5p and TNFAIP8. Capan-1 was divided into miR-NC group， miR-671-5p group， si-NC group， si-TNFAIP8 group， miR-671- 5p+pcDNA group and miR-671-5p+pcDNA-TNFAIP8 group. Cell viability was detected by MTT assay ； the apoptosis was detected by flow cytometry ；the expression levels of cyclin D1， p21， B cell lymphoma/leukemia-2 （ Bcl-2） and Bcl-2 associated X protein （ Bax） were detected by Western blotting. Results Compared with adjacent tissues， the expression of miR-617-5p was significantly decreased in pancreatic cancer tissues， and the expression of TNFAIP8 was significantly increased. miR-671-5p targeted and negatively regulated the expression of TNFAIP8. Compared with the miR-NC group， the viability of capan-1 cells in the miR-671-5p group was significantly decreased， the apoptosis rate was significantly increased， the expression levels of cyclin D1 and Bcl-2 were significantly decreased， and the expression levels of p21 and Bax were significantly increased ； compared with si-NC group， the viability of capan-1 cells in si-TNFAIP8 group was significantly decreased， the apoptosis rate was significantly increased， the expression of cyclin D1 and Bcl-2 was significantly decreased， and the expression levels of p21 and Bax were significantly elevated ； compared with miR-671-5p+pcDNA group， the viability of capan-1 cells in miR-671-5p+pcDNATNFAIP8 group was significantly increased， and the apoptosis rate was significantly decreased， the expression level of cyclin D1 and Bcl-2 was significantly increased， and the expression levels of p21 and Bax were significantly decreased. Conclusion miR-671-5p inhibits proliferationand promotes apoptosis of pancreatic cancer cells by down-regulating TNFAIP8. Up-regulation of miR-671-5p is a potential therapeutic target for pancreatic cancer.