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目的:研究胰腺癌患者血清微RNA-28-5p(miR-28-5p)的表达情况及其对人胰腺癌细胞增殖、迁移及自噬基因BECN1表达的影响。方法:纳入我院消化科门诊及病房确诊胰腺癌的60例患者为试验组,60名健康成人为对照组,提取外周血血清,TaqMan实时定量(qRT)-PCR实验检测血清中miR-28-5p的表达水平;Targetscan预测自噬基因BECN1为miR-28-5p的靶基因,采用蛋白质印迹、荧光素酶报告基因检测(luciferase reporter assay)验证BECN1为miR-28-5p的直接靶基因;细胞计数试剂盒8(CCK-8)法和Transwell试验检测miR-28-5p对人胰腺癌细胞增殖、迁移能力的影响。结果:与正常健康人相比,胰腺癌患者血清中miR-28-5p的表达水平明显降低(39.90±1.64比80.50±1.40,P<0.05);BECN1为miR-28-5p的直接靶基因;增高miR-28-5p的表达使得人胰腺癌细胞的增殖和迁移能力明显降低(1.69±0.07比2.79±0.06,P<0.05;15.89±0.79比29.78±0.60,P<0.05);抑制miR-28-5p则获得相反的效果(2.99±0.12比1.98±0.21,P<0.05;60.40±2.15比38.40±1.09,P<0.05)。结论:胰腺癌患者血清中miR-28-5p较正常人明显降低,miR-28-5p可抑制胰腺癌细胞的增殖和迁移、促进BECN1的表达从而发挥抗肿瘤作用。
Objective: To investigate the expression of microRNA-28-5p (miR-28-5p) and its effect on the proliferation and migration of human pancreatic cancer cells and the expression of autophagy gene BECN1 in patients with pancreatic cancer. Methods: Sixty patients with gastrointestinal cancer and ward diagnosed as pancreatic cancer in our hospital were selected as test group and 60 healthy adults as control group. Peripheral blood serum was extracted and real-time quantitative PCR (qRT) -PCR was used to detect the expression of miR-28- Targeted prediction of autophagy gene BECN1 as miR-28-5p target gene, using Western blot, luciferase reporter assay (luciferase reporter assay) to verify that BECN1 is a direct target of miR-28-5p; Cells The effects of miR-28-5p on the proliferation and migration of human pancreatic cancer cells were detected by counting kit 8 (CCK-8) assay and Transwell assay. Results: The expression of miR-28-5p in the serum of patients with pancreatic cancer was significantly lower than that in healthy controls (39.90 ± 1.64 vs 80.50 ± 1.40, P <0.05). BECN1 was a direct target of miR-28-5p. Increased miR-28-5p expression significantly reduced the proliferation and migration ability of human pancreatic cancer cells (1.69 ± 0.07 vs 2.79 ± 0.06, P <0.05; 15.89 ± 0.79 vs. 29.78 ± 0.60, P <0.05) -5p yielded the opposite effect (2.99 ± 0.12 vs 1.98 ± 0.21, P <0.05; 60.40 ± 2.15 vs. 38.40 ± 1.09, P <0.05). Conclusion: The serum level of miR-28-5p in pancreatic cancer patients is significantly lower than that in normal people. MiR-28-5p can inhibit the proliferation and migration of pancreatic cancer cells and promote the expression of BECN1 to exert anti-tumor effect.