Epidermal growth factor-containing fibulin-like extracellular matrix protein 1 (EFEMP1) suppressed the growth of hepatocellular carcinoma cells by promoting Semaphorin 3B(SEMA3B)
Epidermal growth factor-containing fibulin-like extracellular matrix protein 1 (EFEMP1) suppressed the growth of hepatocellular carcinoma cells by promoting Semaphorin 3B(SEMA3B)
复制标题
含有fibulin样细胞外基质蛋白1（EFEMP1）的表皮生长因子通过促进Semaphorin 3B（SEMA3B）抑制肝细胞癌细胞的生长
DOI：
10.1002/cam4.2144
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发表时间：
2019-06-01
期刊：
影响因子：
4
通讯作者：
Lu, Lungen
中科院分区：
文献类型：
--
作者：
Hu, Jiangfeng;Duan, Bensong;Lu, Lungen
目的含有表皮生长因子的纤维蛋白样细胞外基质蛋白1(EFEMP1)已被发现参与多种癌症的发生和发展。 EFEMP1与肝细胞癌（HCC）发生发展的关系及分子机制尚不完全清楚。方法采用实时聚合酶链反应（PCR）和组织芯片检测EFEMP1在HCC细胞系和组织中的表达。甲基化特异性 PCR 测定用于测量 HCC 细胞系和组织中 EFEMP1 的甲基化水平。为了研究EFEMP1对细胞功能的影响，Huh7和HepG2被表达EFEMP1的慢病毒颗粒感染。采用MTT法和集落形成实验检测EFEMP1对细胞增殖的影响。采用Annexin-VAPC/7-AAD双联检测EFEMP1对细胞凋亡的影响。为了进一步检测EFEMP1对体内HCC发生发展的影响，我们进行了裸鼠成瘤实验。采用基因芯片检测过表达EFEMP1的Huh7和HepG2的表达谱。为了进一步筛选差异，进行了GO分析和pathway分析。为了研究SEMA3B的作用，使用特异性siRNA抑制SEMA3B的表达。采用卡方检验和秩和检验分析EFEMP1表达与HCC临床特征的关系。结果研究发现EFEMP1在HCC细胞系和HCC组织中的表达显着降低。 EFEMP1的表达水平与HCC患者的TNM（肿瘤范围、淋巴结扩散程度、有无转移）分期及预后相关。蛋白表达的降低提示患者预后较差，EFEMP1蛋白水平可能是HCC患者预后的独立因素。启动子甲基化可能是EFEMP1抑制的原因之一。 EFEMP1能够抑制HCC细胞增殖，促进HCC细胞凋亡，从而调控HCC的发生发展。 EFEMP1主要通过线粒体凋亡途径促进HCC细胞凋亡。 EFEMP1可能通过Axon引导通路中的SEMA3B基因抑制HCC细胞的增殖。结论 总之，我们的研究揭示了 EFEMP1 对 HCC 细胞增殖和凋亡的调节。 EFEMP1可能通过促进SEMA3B抑制HCC细胞的生长。
Aim Epidermal growth factor-containing fibulin-like extracellular matrix protein 1(EFEMP1) has been found to be involved in the occurrence and development of many cancers. The relationship between EFEMP1 and the development of hepatocellular carcinoma (HCC) and the molecular mechanism are not fully understood. Methods Real-time polymerase chain reaction (PCR) and tissue microarray were used to detect the expression of EFEMP1 in HCC cell lines and tissue. Methylation-specific PCR assay was used to measure the methylation level of EFEMP1 in HCC cell lines and tissue. To study the function of EFEMP1 on cell function, Huh7 and HepG2 were infected with lentiviral particles expressing EFEMP1. MTT assay and colony formation assay were used to examine the effect of EFEMP1 on cell proliferation. Annexin-VAPC/7-AAD double were used to detect the effect of EFEMP1 on cell apoptosis. To further detect the effect of EFEMP1 on the development of HCC in vivo, we performed the tumor formation experiment in nude mice. Gene chip was used to detect the expression profile of Huh7 and HepG2 overexpressing EFEMP1. To further screen out the differences, GO analysis and pathway analysis were performed. To study the effects of SEMA3B, specific siRNA was used to inhibit the expression of SEMA3B. Chi-squared test and rank sum test were used to analyze the relationship between EFEMP1 expression and HCC clinical characteristic. Results The study found that the expression of EFEMP1 was significantly decreased in HCC cell lines and HCC tissues. The expression level of EFEMP1 was related to the TNM (the extent of the tumor, the extent of spread to the lymph nodes, the presence of metastasis) stage and the prognosis of patients with HCC. The decrease of protein expression suggested that the patient prognosis was worse, and the protein level of EFEMP1 may be an independent factor in the prognosis of HCC patients. Promoter methylation may be one of the reasons for EFEMP1 inhibition. EFEMP1 could inhibit the proliferation of HCC cells and promoted the apoptosis of HCC cells to regulate the development of HCC. And EFEMP1 promoted the apoptosis of HCC cells mainly through the mitochondrial apoptosis pathway. EFEMP1 may inhibit the proliferation of HCC cells through the SEMA3B gene in the Axon guidance pathway. Conclusion In summary, our research revealed the regulation of EFEMP1 on cell proliferation and apoptosis in HCC. EFEMP1 may suppress the growth of HCC cells by promoting SEMA3B.