Mutation analysis of BRAF and KIT in circulating melanoma cells at the single cell level.
Mutation analysis of BRAF and KIT in circulating melanoma cells at the single cell level.
复制标题
DOI：
10.1038/bjc.2012.12
复制
发表时间：
2012-02-28
影响因子：
8.8
通讯作者：
Okuyama, R.
中科院分区：
文献类型：
--
作者：
Sakaizawa, K.;Goto, Y.;Kiniwa, Y.;Uchiyama, A.;Harada, K.;Shimada, S.;Saida, T.;Ferrone, S.;Takata, M.;Uhara, H.;Okuyama, R.
治疗黑色素瘤的分子靶向疗法的可用性强调了识别目标基因突变的必要性，如BRAF和KIT。循环肿瘤细胞(CTC)存在于相当大比例的癌症患者的外周血中。利用高分子黑色素瘤相关抗原(HMW-MAA)在黑色素瘤中的高选择性表达，从外周血中分离黑色素瘤细胞。用免疫磁珠分离HMW-MAA阳性细胞。去除CD45+细胞后，用MART-1和Gp100特异性抗体(HMW-MAA+、CD45−、MART-1/GP100+)进行鉴定。然后对单个、分离的CTC进行BRAF和KIT突变分析。从11例患者的血液中分离出CTC(HMW-MAA+、CD45Gp100+、MART-1/gp100+)，分别对9例和4例患者进行BRAF和KIT测序。CTC中鉴定的BRAF序列与3名患者自体黑色素瘤中鉴定的BRAF序列不一致，3名患者的KIT序列不一致。此外，在一名患者中发现了多克隆的BRAF突变，在另一名患者中发现了伴随的BRAF和KIT突变。黑色素瘤细胞呈克隆性异质性。因此，CTC基因分型可能是分子靶向治疗成功的关键。
The availability of molecular-targeted therapies for the treatment of melanoma has emphasised the need to identify mutations in target genes such as BRAF and KIT. Circulating tumour cells (CTC) are present in the peripheral blood of a significant proportion of cancer patients. High molecular weight melanoma-associated antigen (HMW-MAA) was used to isolate melanoma cells from peripheral blood as it is selectively expressed at high levels on melanomas. The HMW-MAA-positive cells were isolated using immunomagnetic beads. After removing CD45+ cells, CTC were identified by staining with MART-1- and gp100-specific antibodies (HMW-MAA+, CD45−, MART-1/gp100+). Single, isolated CTC were then subjected to BRAF and KIT mutational analysis. CTC (HMW-MAA+, CD45−, MART-1/gp100+) were isolated from the blood of 11 patients and BRAF and KIT were sequenced in nine and four patients, respectively. The BRAF sequences identified in the CTC were inconsistent with those identified in autologous melanoma tumours in three patients and the KIT sequences were inconsistent in three patients. In addition, polyclonal BRAF mutations were identified in one patient and concomitant mutations in BRAF and KIT were identified in another patient. Melanoma cells show clonal heterogeneity. Therefore, CTC genotyping may be crucial for successful molecular-targeted therapy.
登录
查看更多内容
影响因子：
64.5
作者：
Heidorn SJ;Milagre C;Whittaker S;Nourry A;Niculescu-Duvas I;Dhomen N;Hussain J;Reis-Filho JS;Springer CJ;Pritchard C;Marais R
通讯作者：
Marais R
影响因子：
10.3
作者：
Lin, Jingrong;Takata, Minoru;Saida, Toshiaki
通讯作者：
Saida, Toshiaki
影响因子：
3.7
作者：
Lanternier, Fanny;Cohen-Akenine, Annick;Palmerini, Fabienne;Feger, Frederic;Yang, Ying;Zermati, Yael;Barete, Stephane;Sans, Beatrix;Baude, Cedric;Ghez, David;Suarez, Felipe;Delarue, Richard;Casassus, Philippe;Bodemer, Christine;Catteau, Adeline;Soppelsa, Frederique;Hanssens, Katia;Arock, Michel;Sobol, Hagay;Fraitag, Sylvie;Canioni, Daniele;Moussy, Alain;Launay, Jean Marie;Dubreuil, Patrice;Hermine, Olivier;Lortholary, Olivier
通讯作者：
Lortholary, Olivier
影响因子：
6.5
作者：
De Giorgi, Vincenzo;Pinzani, Pamela;Massi, Daniela
通讯作者：
Massi, Daniela
影响因子：
9.3
作者：
Koyanagi, K;Kuo, C;Hoon, DSB
通讯作者：
Hoon, DSB