Yang Guang-Yun, Zhang Ai-Qun, Wang Jing, Li Chong-Hui, Wang Xian-Qiang, Pan Ke, Zhou Cheng, Dong Jia-Hong
Department of Hepatobiliary Surgery, PLA General Hospital and Institute of Hepatobiliary of Chinese PLA, Beijing, China.
Cell Biochem Funct. 2016 Jun;34(4):274-85. doi: 10.1002/cbf.3189.
We aimed to elucidate the effects of hepatoma-derived growth factor (HDGF) on growth and metastasis of hepatocellular carcinoma (HCC) cells. Tissue microarrays with 236 HCC specimens and 18 extrahepatic metastases were utilized to detect the HDGF expression by immunohistochemistry. Meanwhile, HDGF expressions in HCC cell lines with different metastatic potentials were examined using immunofluorescence staining, real-time PCR and western blotting. After HDGF silencing, the growth and metastatic potentials of HCC cells were evaluated by soft agar assay, invasion assay, together with tumorigenicity assay in nude mice. The gelatin zymography was performed by detecting MMP-2 and MMP-9 levels. Additionally, western blotting was conducted to determine the levels of total and phosphorylated ERK1/2, JNK, p38 and Akt. The results showed that HDGF was overexpressed in HCC metastasis tumour, and the expression increased with the differentiation degree of tumours (Grade I 44.0%, Grade II 48.4% and Grade III 65.6%). Consistently, HDGF levels were positively associated with the metastatic capability of HCC cells (MHCC97L < MHCC97H < HCCLM3). The growth and metastasis were suppressed by HDGF-siRNA. Gelatinolytic activities were enhanced in the three metastatic HCC cell lines, but had no significant difference among them. The tumourigenicity and metastatic capability of HCCLM3 cells in nude mice were inhibited after silencing HDGF. Meanwhile, HDGF-siRNA specifically suppressed the total and phosphorylated protein levels of ERK1/2, while not JNK, p38 and Akt. In conclusion, HDGF was overexpressed in HCC patients and cells, and HDGF might be closely correlated with HCC metastasis via regulating ERK signalling pathway. Copyright © 2016 John Wiley & Sons, Ltd.
我们旨在阐明肝癌衍生生长因子（HDGF）对肝细胞癌（HCC）细胞生长和转移的影响。利用含有236例HCC标本和18例肝外转移灶的组织芯片，通过免疫组织化学检测HDGF表达。同时，采用免疫荧光染色、实时PCR和蛋白质印迹法检测不同转移潜能HCC细胞系中的HDGF表达。HDGF沉默后，通过软琼脂试验、侵袭试验以及裸鼠致瘤性试验评估HCC细胞的生长和转移潜能。通过检测基质金属蛋白酶-2（MMP-2）和基质金属蛋白酶-9（MMP-9）水平进行明胶酶谱分析。此外，进行蛋白质印迹法以测定总ERK1/2、磷酸化ERK1/2、JNK、p38和Akt的水平。结果显示，HDGF在HCC转移瘤中过表达，且表达随肿瘤分化程度增加（I级44.0%，II级48.4%，III级65.6%）。一致地，HDGF水平与HCC细胞的转移能力呈正相关（MHCC97L ＜ MHCC97H ＜ HCCLM3）。HDGF-siRNA抑制了生长和转移。三种转移性HCC细胞系中的明胶酶活性增强，但它们之间无显著差异。HDGF沉默后，抑制了HCCLM3细胞在裸鼠中的致瘤性和转移能力。同时，HDGF-siRNA特异性抑制ERK1/2的总蛋白和磷酸化蛋白水平，而对JNK、p38和Akt无影响。总之，HDGF在HCC患者和细胞中过表达，且HDGF可能通过调节ERK信号通路与HCC转移密切相关。版权所有© 2016约翰威立父子有限公司。
