Mortalin stabilizes CD151-depedent tetraspanin-enriched microdomains and implicates in the progression of hepatocellular carcinoma
Mortalin stabilizes CD151-depedent tetraspanin-enriched microdomains and implicates in the progression of hepatocellular carcinoma
复制标题
Mortalin 稳定 CD151 依赖的富含四跨膜蛋白的微结构域并参与肝细胞癌的进展
DOI：
10.7150/jca.36301
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发表时间：
2019-01-01
影响因子：
3.9
通讯作者：
Shi, Guo-Ming
中科院分区：
文献类型：
--
作者：
Liu, Li-Xin;Lu, Jia-Cheng;Shi, Guo-Ming
背景资料：我们以前的研究表明，四跨膜蛋白CD 151与肝细胞癌（HCC）的进展有关，主要取决于与分子伴侣（包括Mortalin）形成功能复合物。在这项研究中，我们研究了死亡蛋白在CD 151依赖性肝癌进展中的作用。研究方法：应用免疫荧光染色、western blot和实时荧光定量聚合酶链反应（qRT-PCR）检测CD 151和Mortalin在4种不同转移能力肝癌细胞系中的表达和定位。采用免疫共沉淀法研究了HCCLM 3细胞中Mortalin与CD 151之间的关系。通过转染技术改变肝癌细胞中CD 151或Mortalin的表达。创伤愈合实验和Transwell实验检测CD 151和Mortalin在细胞迁移和侵袭中的作用。分析187例HCC组织中CD 151和Mortalin的表达及其与预后的关系。结果如下：肝癌细胞中Mortalin的表达与其转移能力呈正相关，其趋势与CD 151的表达一致。免疫荧光染色显示Mortalin主要定位于细胞浆，而CD 151主要定位于细胞浆和细胞膜。co-IP显示Mortalin与CD 151形成复合物。Mortalin的下调诱导了CD 151蛋白的中度降低，但不诱导CD 151 mRNA的降低，而CD 151的抑制并不影响Mortalin在蛋白和mRNA水平上的表达。Mortalin的干预可显著抑制CD 151高表达的肝癌细胞的侵袭和迁移，并部分恢复CD 151过表达诱导的肝癌细胞的侵袭和迁移。临床上，Mortalin高表达与HCC的恶性表型相关，如微血管浸润（p=0.017）和肿瘤直径（p=0.001）。高表达Mortalin的HCC患者有CD 151高表达的倾向。CD 151高表达的HCC患者预后较差，且与Mortalin呈依赖关系。结论：Mortalin可能稳定了肝癌细胞中依赖于CD 151的四跨膜蛋白富集微区的结构，并参与了肝癌的发生、发展。
Background: Our previous studies showed that tetraspanin CD151 was implicated in the progression of hepatocellular carcinoma (HCC), mainly depending on the formation of functional complexes with molecular partners, including Mortalin. In this study, we investigate the role of mortalin in CD151-depedent progression of HCCs. Methods: Immunofluorescent staining, western blot and quantitative real-time polymerase chain reaction (qRT-PCR) were used to investigate the expression and location of CD151 and Mortalin in four HCC cell lines with different metastatic ability. The relationship between Mortalin and CD151 was investigated in HCCLM3 cells using co-immunoprecipitation. CD151 or Mortalin expression in HCC cells were modified by transfection technology. Wound-healing assay and Transwell assay were used to assay the role of CD151 and Mortalin in cell migration and invasion. The expression and prognostic implication of CD151 and Mortalin in 187 cases of HCCs were analyzed. Results: Expression of Mortalin in HCC cells was positive related to their metastatic ability and its tendency was in line with the expression of CD151. Immunofluorescent staining showed that Mortalin was located in cytoplasm, while positive staining for CD151 was observed in cytoplasm and membrane of HCC cells. co-IP revealed that Mortalin formed a complex with CD151. Down-regulation of Mortalin induced a moderate decreased CD151 protein, but not CD151 mRNA, while inhibition of CD151 did not influence the expression of Mortalin at the level of both protein and mRNA. Interference of Mortalin significantly inhibited the invasion and migration of HCC cells with high CD151 expression and partially restored the invasion and migration of HCC cells induced by CD151 over-expression. Clinically, high Mortalin expression correlated with malignant phenotype of HCC, such as microvascular invasion (p=0.017) and tumor diameter (p=0.001). HCC patients expressing high Mortalin were tend to have higher expression of CD151. HCC patients expressing high level of CD151 showed the poorer prognosis in a Mortalin-dependent manner. Conclusions: Mortalin maybe stabilize of the structure of CD151-dependent tetraspanin-enriched microdomains and implicate in the progression of HCC.