Cytokeratin expression in malignant melanoma: potential application of in-situ hybridization analysis of mRNA
Cytokeratin expression in malignant melanoma: potential application of in-situ hybridization analysis of mRNA
复制标题
恶性黑色素瘤中细胞角蛋白的表达：mRNA 原位杂交分析的潜在应用
DOI：
10.1097/cmr.0b013e3283252feb
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发表时间：
2009-04-01
影响因子：
2.2
通讯作者：
Zhou, Qiao
中科院分区：
文献类型：
--
作者：
Chen, Ni;Gong, Jing;Zhou, Qiao
偶尔有报道指出细胞角蛋白（CK）蛋白在恶性黑色素瘤（MM）中表达（主要通过免疫组织化学方法），并提示其与不良临床参数相关。然而，细胞角蛋白的mRNA表达及其在MM中的临床病理意义尚未得到专门评估。我们研究了9种细胞角蛋白在黑色素瘤细胞系和组织中的mRNA和蛋白表达，特别是CK18 mRNA表达的预后意义。对黑色素瘤细胞系A375、A875、M14和SK - MEL - 1进行了逆转录（RT）- PCR（检测CK6 - 10、14和18 - 20）、原位杂交（ISH）（检测CK18）以及蛋白质印迹法（检测CK18和广谱细胞角蛋白AE1/AE3）。通过ISH和免疫组织化学方法对80例MM组织样本进行CK18表达分析。通过RT - PCR在1 - 4种黑色素瘤细胞系中检测到CK6 - 8、10、14、18和19（但不是CK9和20）的mRNA。通过RT - PCR、ISH和蛋白质印迹法在所有4种细胞系中均检测到CK18。在30例原发性皮肤黑色素瘤中有3例（10.0%）、25例原发性黏膜黑色素瘤中有10例（40.0%）以及25例转移性黑色素瘤中有12例（48.0%）CK18 mRNA的ISH检测呈阳性（总体阳性率：80例中有25例，31.3%）。在80例MM组织样本中仅有8例（10.0%）局部观察到CK18免疫染色，而AE1/AE3免疫染色完全为阴性。值得注意的是，通过单变量分析（P < 0.001）和多变量分析（相对危险度 = 5.430，95%置信区间2.246 - 13.128，P < 0.001），CK18 mRNA的ISH阳性（而非蛋白免疫组织化学）与较差的预后相关。在三分之一的黑色素瘤组织样本中可检测到CK18 mRNA，它是一个不良预后因素。在分析MM中CK18表达方面，ISH优于免疫组织化学。《黑色素瘤研究》19:87 - 93（c）2009年，威科集团 | 利平科特·威廉姆斯和威尔金斯出版社。
Occasional reports indicated cytokeratin (CK) protein expression (mainly by immunohistochemistry) in malignant melanoma (MM) and suggested an association with unfavorable clinical parameters. However, the mRNA expression of CK and its clinicopathologic significance in MM has not been specifically evaluated. We investigated the mRNA and protein expression of nine CKs in melanoma cell lines and tissues, in particular the prognostic significance of CK18 mRNA expression. Reverse transcription (RT)-PCR (CK6–10, 14 and 18–20), in-situ hybridization (ISH) (CK18), and western blotting (CK18 and pan-cytokeratin AE1/AE3) were performed on MM cell lines A375, A875, M14, and SK-MEL-1. Eighty MM tissue samples were analyzed by ISH and immunohistochemistry for CK18 expression. The mRNA of CK6–8, 10, 14, 18, and 19 (but not CK9 and 20) was detected in one to four of the melanoma cell lines by RT-PCR. CK18 was detected in all four cell lines by RT-PCR, ISH, and western blotting. CK18 mRNA ISH was positive in three of 30 (10.0%), 10 of 25 (40.0%), and 12 of 25 (48.0%) of primary cutaneous, primary mucosal, and metastatic melanomas, respectively (overall positivity: 25 of 80, 31.3%). CK18 immunostaining was only observed focally in eight of 80 (10.0%) of MM tissue samples, and AE1/AE3 immunostaining was altogether negative. Significantly, CK18 mRNA ISH positivity (but not protein immunohistochemistry) was associated with poorer prognosis by both univariate analysis (P<0.001) and multivariate analysis (relative risk=5.430, 95% confidence interval 2.246–13.128, P<0.001). CK18 mRNA could be identified in one-third of melanoma tissue samples and is an adverse prognostic factor. ISH is superior to immunohistochemistry for analyzing CK18 expression in MM.