Department of Pathology, Tianjin Cancer Institute and Hospital, Tianjin Medical University, Tianjin, People's Republic of China.
J Exp Clin Cancer Res. 2010 Aug 11;29(1):109. doi: 10.1186/1756-9966-29-109.
The incidence of malignant melanoma (MM) was occurring at a faster rate than for most neoplasm worldwide, and melanoma metastasis is still the most formidable problem. So it is necessarily to find some biomarkers associated with melanoma metastasis.
In our study, 8 spontaneous lung metastatic mice models were created by B16F10 subcutaneously transplantation. The differential protein profiles of two kinds of subcutaneous transplanted tumor tissues, which was parental B16F10 (B16 group) and corresponding lung metastases (B16M group) were detected by two-dimensional differential in-gel electrophoresis (2D-DIGE) combined with matrix-assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF/MS). Western blotting was used to validate the results, and the clinical significance of individual protein was detected furtherly in a set of human samples.
In this study, thirty proteins were found to be differentially expressed (ratio > 2 or < -2, P < 0.01) and thirteen of them were identified by MS. Highly expressed proteins in B16M group included cytoskeleton/structure proteins (vimentin, gamma-actin, beta-actin, laminin binding protein), the chaperone family of proteins (heavy-chain binding protein, Bip), immunoproteasome assembly (proteasome activator REG alpha) and others involved in glycolysis activity (PGK1, enolase, TPI, human skeletal muscle GAPDH) and protein transport (myoglobin). Vimentin was significantly up-regulated in B16M group compared with B16 group which was validated by western blotting. Immunohistochemistry was performed in a set of clinical samples, the results showed that over-expression of vimentin was frequently observed in primary melanoma patients with hematogenous metastasis (P < 0.05), not associated with lymph node metastasis (P > 0.05). The presence of TNM stage was a independent indicator of poor prognosis for melanoma patients (P = 0.004).
The aberrant immunohistochemical expression of vimentin in primary melanoma tissues may help to call attention for patients with high risk of hematogenous metastasis. That might be as a novel metastatic indicator for melanoma. In a word, vimentin is not only the diagnostic marker but also the hematogenous metastasis predictor for melanomas clinically.
恶性黑色素瘤(MM)的发病率比世界上大多数肿瘤都要快,黑色素瘤转移仍然是最棘手的问题。因此,有必要寻找一些与黑色素瘤转移相关的生物标志物。
我们通过 B16F10 皮下移植建立了 8 例自发性肺转移小鼠模型。通过二维差异凝胶电泳(2D-DIGE)联合基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF/MS)检测两种皮下移植肿瘤组织的差异蛋白谱,即亲本 B16F10(B16 组)和相应的肺转移(B16M 组)。Western blot 进一步验证了结果,并在一组人类样本中检测了单个蛋白的临床意义。
本研究发现 30 种蛋白表达差异(比值>2 或<-2,P<0.01),其中 13 种蛋白通过 MS 鉴定。B16M 组高表达的蛋白包括细胞骨架/结构蛋白(波形蛋白、γ-肌动蛋白、β-肌动蛋白、层粘连蛋白结合蛋白)、伴侣蛋白家族(重链结合蛋白、Bip)、免疫蛋白酶体组装(蛋白酶体激活剂 REG alpha)和其他参与糖酵解活性(PGK1、烯醇酶、TPI、人骨骼肌 GAPDH)和蛋白质运输(肌红蛋白)的蛋白。Western blot 验证了 B16M 组中波形蛋白的表达明显上调。在一组临床样本中进行了免疫组织化学检测,结果显示,在发生血行转移的原发性黑色素瘤患者中,波形蛋白的过度表达经常观察到(P<0.05),与淋巴结转移无关(P>0.05)。TNM 分期的存在是黑色素瘤患者预后不良的独立指标(P=0.004)。
原发性黑色素瘤组织中波形蛋白异常的免疫组化表达可能有助于引起对高血行转移风险患者的关注。这可能是黑色素瘤的一个新的转移指标。总之,波形蛋白不仅是黑色素瘤的诊断标志物,也是黑色素瘤血行转移的预测标志物。