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目的:研究脑源性神经营养因子(brain-derived neurotrophic factor,BDNF)及其受体原肌球蛋白相关激酶B(Tropomyosin-related kinase B,TrkB)在人肝细胞癌(hepatocellular carcinoma,HCC)中的表达情况,并探讨2者在HCC发生、发展中的作用。方法:采用蛋白质印迹法检测BDNF和TrkB蛋白在30例HCC和癌旁组织中的表达情况。采用ELISA法检测BDNF在人HCC细胞系97-H培养液上清中的浓度;FCM和Transwell小室法分别检测抗BDNF抗体或TrkB激酶活性抑制剂K252a对细胞凋亡和侵袭的影响。结果:30例配对组织标本中,BDNF和TrkB在HCC组织中的表达水平高于癌旁组织(P均<0.05)。BDNF在97-H细胞培养液上清中的表达量为(119.08±6.21)pg/mL。抗BDNF抗体或K252a都能有效诱导97-H细胞的凋亡,并抑制细胞的侵袭能力。结论:BDNF/TrkB可能对HCC细胞的存活和侵袭具有重要的支持作用,并促进HCC的发生及发展。
Objective: To investigate the expression of brain-derived neurotrophic factor (BDNF) and its receptor Tropomyosin-related kinase B (TrkB) in human hepatocellular carcinoma (HCC) And to explore the role of the two in HCC occurrence and development. Methods: Western blotting was used to detect the expression of BDNF and TrkB protein in 30 cases of HCC and adjacent normal tissues. The concentration of BDNF in the supernatant of human HCC cell line 97-H was detected by ELISA. The effect of anti-BDNF antibody or TrkB kinase inhibitor K252a on the apoptosis and invasion was detected by FCM and Transwell chamber assay. Results: The expression of BDNF and TrkB in HCC tissues was higher than that in adjacent tissues (P <0.05). The expression of BDNF in 97-H cell culture supernatant was (119.08 ± 6.21) pg / mL. Anti-BDNF antibody or K252a can effectively induce the apoptosis of 97-H cells and inhibit the invasion ability of cells. Conclusion: BDNF / TrkB may play an important supporting role in the survival and invasion of HCC cells and promote the occurrence and development of HCC.