Huang Pei-Yu, Hong Ming-Huang, Zhang Xing, Mai Hai-Qiang, Luo Dong-Hua, Zhang Li
State Key Laboratory of Oncology in South China, Guangzhou, Guangdong 510060, PR China.
Chin J Cancer. 2010 Feb;29(2):131-5. doi: 10.5732/cjc.009.10411.
We previously reported that C-KIT overexpression and mutation exist in biopsy samples of nasopharyngeal carcinoma (NPC). Yet whether Imatinib had an inhibitory effect on the proliferation of NPC in vitro was still unknown. So, this study examined whether sensitivities to Imatinib of other cell lines are different and whether C-KIT expression and mutations exist, to analyze the correlations between them.
The expression of C-KIT in NPC cell lines, including CNE-1, CNE-2, Hone-1, C-666, SUNE-1, 5-8F, and nasopharyngeal epithelial (NPE) cell line NP-69, were detected by Western blot. Direct sequencing of polymerase chain reaction (PCR) products was performed to analyze the sequences of C-KIT from the above-mentioned cell lines. Inhibitory effects on proliferation by Imatinib on these cell lines were determined by CCK-8 assay. Pearson product moment correlation and t test were used to analyze the correlation betweeen C-KIT overexpression, C-KIT gene mutation, and the inhibitory effect of Imatinib.
Compared with NPE cell line NP-69, NPC cell lines CNE-1, CNE-2, Hone-1, C-666, SUNE-1, and 5-8F had significantly higher levels of C-KIT expression. Heterozygous IVS17+78T>C were found in CNE-1, CNE-2, Hone-1, and NP-69 cell lines, homozygous IVS17+78T>C was found in C-666, and no mutation was found in SUNE-1 or 5-8F. Imatinib had a dose-dependent inhibitory effect on proliferation for CNE-1, CNE-2, Hone-1, C-666, SUNE-1, and 5-8F. No significant correlation between the inhibitory effects of Imatinib, C-KIT overexpression, or C-KIT mutation was found.
C-KIT overexpression and intron mutation were found in NPC cell lines and Imatinib had a dose-dependent inhibitory effect on proliferation for NPC cell lines, yet no significant correlation between C-KIT overexpression, C-KIT mutation, or the inhibitory effect of Imatinib was found.
我们之前报道过,在鼻咽癌(NPC)活检样本中存在C-KIT过表达和突变。然而,伊马替尼在体外对NPC增殖是否具有抑制作用仍不清楚。因此,本研究检测了其他细胞系对伊马替尼的敏感性是否不同以及是否存在C-KIT表达和突变,以分析它们之间的相关性。
通过蛋白质免疫印迹法检测NPC细胞系(包括CNE-1、CNE-2、Hone-1、C-666、SUNE-1、5-8F)及鼻咽上皮(NPE)细胞系NP-69中C-KIT的表达。对上述细胞系的C-KIT进行聚合酶链反应(PCR)产物直接测序以分析其序列。采用CCK-8法检测伊马替尼对这些细胞系增殖的抑制作用。使用Pearson积矩相关分析和t检验分析C-KIT过表达、C-KIT基因突变与伊马替尼抑制作用之间的相关性。
与NPE细胞系NP-69相比,NPC细胞系CNE-1、CNE-2、Hone-1、C-666、SUNE-1和5-8F的C-KIT表达水平显著更高。在CNE-1、CNE-2、Hone-1和NP-69细胞系中发现杂合子IVS17+78T>C,在C-666中发现纯合子IVS17+78T>C,而在SUNE-1或5-8F中未发现突变。伊马替尼对CNE-1、CNE-2、Hone-1、C-666、SUNE-1和5-8F的增殖具有剂量依赖性抑制作用。未发现伊马替尼的抑制作用、C-KIT过表达或C-KIT突变之间存在显著相关性。
在NPC细胞系中发现C-KIT过表达和内含子突变,且伊马替尼对NPC细胞系的增殖具有剂量依赖性抑制作用,但未发现C-KIT过表达、C-KIT突变与伊马替尼抑制作用之间存在显著相关性。