鉴定8号染色体p臂上的MSRA基因作为人乙型肝炎病毒阳性肝细胞癌的候选转移抑制基因。
Identification of MSRA gene on chromosome 8p as a candidate metastasis suppressor for human hepatitis B virus-positive hepatocellular carcinoma.
作者信息
Lei Ke-Feng, Wang Yan-Fang, Zhu Xiao-Qun, Lu Peng-Cheng, Sun Bing-Sheng, Jia Hu-Liang, Ren Ning, Ye Qing-Hai, Sun Hui-Chuan, Wang Lu, Tang Zhao-You, Qin Lun-Xiu
机构信息
Liver Cancer Institute and Zhongshan Hospital, Institutes of Biomedical Science, Fudan University, Shanghai, China.
出版信息
BMC Cancer. 2007 Sep 4;7:172. doi: 10.1186/1471-2407-7-172.
BACKGROUND
The prognosis of patients with hepatocellular carcinoma (HCC) still remains very dismal, which is mainly due to metastasis. In our previous studies, we found that chromosome 8p deletions might contribute to metastasis of HCC. In this study, we aimed to identify the candidate metastatic suppressor gene on chromosome 8p.
METHODS
Oligo-nucleotide microarrays which included 322 genes on human chromosome 8p were constructed to analyze the difference in gene expression profiles between HCC tissues with and without metastasis. The leading differentially expressed genes were identified and selected for further analysis by real-time PCR and Western blotting. Recombinant expression plasmid vectors for each target gene were constructed and transfected into HCC cells and its in vitro effects on proliferation and invasion of HCC cells were also investigated.
RESULTS
Sixteen leading differentially expressed genes were identified from the HCC tissues with metastasis compared with those without metastasis (p < 0.01, q < 16 %). Among of the 10 significantly down-regulated genes in HCC with metastasis, methionine sulfoxide reductase A (MSRA) had the lowest p value and false discovery rate (FDR), and was considered as a potential candidate for metastasis suppressor gene. Real-time PCR and Western blotting confirmed that the mRNA and protein expression levels of MSRA were significantly decreased in HCC with metastasis compared with those without metastasis (p < 0.001), and MSRA mRNA level in HCCLM6 cells (with high metastatic potential) was also much lower than that of other HCC cell lines. Transfection of a recombinant expression plasmid vector and overexpression of MSRA gene could obviously inhibit cell colony formation (4.33 +/- 2.92 vs. 9.17 +/- 3.38, p = 0.008) and invasion (7.40 +/- 1.67 vs. 17.20 +/- 2.59, p= 0.0001) of HCCLM6 cell line.
CONCLUSION
MSRA gene on chromosome 8p might possess metastasis suppressor activity in HCC.
背景
肝细胞癌（HCC）患者的预后仍然非常严峻，这主要是由于转移所致。在我们之前的研究中，我们发现8号染色体短臂缺失可能与HCC转移有关。在本研究中，我们旨在鉴定8号染色体短臂上的候选转移抑制基因。
方法
构建包含人类8号染色体短臂上322个基因的寡核苷酸微阵列，以分析有转移和无转移的HCC组织之间基因表达谱的差异。鉴定出主要的差异表达基因，并通过实时PCR和蛋白质印迹法进行进一步分析。构建每个靶基因的重组表达质粒载体，并转染到HCC细胞中，同时研究其对HCC细胞增殖和侵袭的体外影响。
结果
与无转移的HCC组织相比，从有转移的HCC组织中鉴定出16个主要的差异表达基因（p < 0.01，q < 16%）。在有转移的HCC中10个显著下调的基因中，甲硫氨酸亚砜还原酶A（MSRA）的p值和错误发现率（FDR）最低，被认为是转移抑制基因的潜在候选者。实时PCR和蛋白质印迹法证实，与无转移的HCC相比，有转移的HCC中MSRA的mRNA和蛋白质表达水平显著降低（p < 0.001），并且HCCLM6细胞（具有高转移潜能）中的MSRA mRNA水平也远低于其他HCC细胞系。转染重组表达质粒载体并过表达MSRA基因可明显抑制HCCLM6细胞系的细胞集落形成（4.33±2.92对9.17±3.38，p = 0.008）和侵袭（7.40±1.67对17.20±2.59，p = 0.0001）。
结论
8号染色体短臂上的MSRA基因可能在HCC中具有转移抑制活性。
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