Ying Hai Yan, Gong Chao Jie, Feng Yi, Jing Da Dao, Lu Lun Gen
Department of Geriatrics, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
Department of Gastroenterology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China.
J Dig Dis. 2017 Jun;18(6):349-358. doi: 10.1111/1751-2980.12486.
To investigate serine protease inhibitor Kazal type 1 (SPINK1) expression and its influence on the prognosis of human hepatocellular carcinoma (HCC) and to explore the underlying molecular mechanisms involved.
Altogether 80 patients with HCC who underwent curative resection were followed up for a median of 58.6 months. SPINK1 expression was detected in the primary HCC samples by immunohistochemistry. Its role in tumor invasion and metastasis was evaluated in vitro by gene silencing using a small interfering RNA-mediated approach, recombinant SPINK1 and U0126 (an inhibitor of MEK/ERK). The proteins in the MEK/ERK signaling pathway were detected by Western blot.
Patients with high SPINK1 expression showed poor overall survival (P = 0.0001) and recurrence-free survival (P = 0.001) compared with those with low SPINK1 expression. The suppression of SPINK1 resulted in reduced cell migration and invasion. SPINK1 overexpression was significantly associated with increased cell migration and invasion in vitro. Furthermore, SPINK1 promoted cancer cells motility and epithelial-mesenchymal transition (EMT) via the mitogen-activated protein kinase kinase (MAPK) and extracellular regulated kinase (ERK) pathway, resulting in increased vimentin expression and decreased E-cadherin expression.
SPINK1 may be an oncogene that induces EMT via the MEK/ERK pathway and is a potential target for HCC therapy.
研究丝氨酸蛋白酶抑制剂Kazal型1（SPINK1）的表达及其对人肝细胞癌（HCC）预后的影响，并探讨其潜在的分子机制。
对80例行根治性切除的HCC患者进行随访，中位随访时间为58.6个月。采用免疫组织化学法检测原发性HCC样本中SPINK1的表达。通过小干扰RNA介导的基因沉默、重组SPINK1和U0126（MEK/ERK抑制剂）在体外评估其在肿瘤侵袭和转移中的作用。采用蛋白质印迹法检测MEK/ERK信号通路中的蛋白。
与低SPINK1表达患者相比，高SPINK1表达患者的总生存期（P = 0.0001）和无复发生存期（P = 0.001）较差。SPINK1的抑制导致细胞迁移和侵袭减少。SPINK1过表达与体外细胞迁移和侵袭增加显著相关。此外，SPINK1通过丝裂原活化蛋白激酶激酶（MAPK）和细胞外调节激酶（ERK）途径促进癌细胞运动和上皮-间质转化（EMT），导致波形蛋白表达增加和E-钙黏蛋白表达减少。
SPINK1可能是一种通过MEK/ERK途径诱导EMT的癌基因，是HCC治疗的潜在靶点。
