TEC酪氨酸蛋白激酶在肝癌组织中高表达并参与肝癌耐药

刘文; 李菲菲; 钟明贵; 刘亚坤; 方皓舒; 汪思应

doi:10.3971/j.issn.1000-8578.2018.18.0107

TEC酪氨酸蛋白激酶在肝癌组织中高表达并参与肝癌耐药

TEC Expresses Highly in Hepatocellular Carcinoma and Is Involved in Drug Resistance

摘要

摘要:
目的研究TEC酪氨酸蛋白激酶在肝癌组织中的表达及其与肝癌耐药之间的关系，为肝癌临床治疗提供线索。
方法 Real-time PCR、Western blot及免疫组织化学法检测癌旁、肝癌组织及肝癌细胞系中TEC mRNA、TEC蛋白表达水平；狭线杂交法检测肝癌组织中TEC mRNA表达水平；MTT检测细胞存活率。构建TEC真核表达载体，经脂质体介导法转染肝癌细胞；RNA干扰技术敲低肝癌细胞中TEC。免疫共沉淀技术检测TEC磷酸化活化水平。
结果 18例肝癌组织中，TEC mRNA和蛋白水平显著高于癌旁组织（P < 0.001）；48例肝癌组织中，TEC高表达组对化疗药物的耐受能力明显高于TEC低表达组；上调TEC表达水平后，HepG2细胞对化疗药物的耐受性增加；用化疗药物处理肝癌细胞株HepG2，其TEC表达水平增加，并伴随有ERK磷酸化水平升高。
结论 TEC在肝癌组织中的高表达促进了肝癌的耐药，且可能与ERK磷酸化水平升高有关。

Abstract:
Objective To investigate the expression level of TEC, a no-receptor tyrosine kinase, in hepatocellular carcinoma(HCC) tissues and its correlation with drug resistance.
Methods The expressions of TEC mRNA and protein in carcinoma tissues, paracancerous tissues and HepG2 cell line were determined by RT-PCR, Western blot, immunohistochemistrical staining and slot blotting. MTT assay was used to detect the activity of HepG2 cells. A eukaryotic expression vector carrying TEC gene was constructed to obtain high expression level of TEC in HCC, and RNA interference was applied to knock down TEC. Phospho-TEC was also analyzed by CO-IP to determine TEC activation.
Results TEC mRNA and protein were both increased significantly in 18 cases of HCC tissues as compared with those in paracancerous tissues(P < 0.001). The high expression of TEC was associated with the increased resistance of 48 cases of HCC tissues to chemotherapeutic drugs; the upregulation of TEC increased the resistance of HepG2 cells to chemotherapeutic drugs. TEC were augmented by the treatment of chemotherapeutics, accompanying with an increase of phospho-ERK.
Conclusion TEC is highly expressed in HCC tissues, which is responsible for the drug resistance of HCC and likely related to increased phospho-ERK.

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