C-KIT overexpression and mutation in nasopharyngeal carcinoma cell lines and reactivity of Imatinib on these cell lines.
C-KIT overexpression and mutation in nasopharyngeal carcinoma cell lines and reactivity of Imatinib on these cell lines.
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DOI：
10.5732/cjc.009.10411
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发表时间：
2010
影响因子：
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通讯作者：
Pei-yu Huang;M. Hong;Xing Zhang;H. Mai;D. Luo;Li Zhang
中科院分区：
文献类型：
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作者：
Pei-yu Huang;M. Hong;Xing Zhang;H. Mai;D. Luo;Li Zhang
背景与目的我们曾报道鼻咽癌组织中存在C-KIT基因的过度表达和突变。但伊马替尼在体外对鼻咽癌细胞增殖是否有抑制作用尚不清楚。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。方法采用Western blot方法检测C-KIT在鼻咽癌细胞系CNE-1、CNE-2、Hone-1、C-666、SUNE-1、5- 8 F和鼻咽上皮细胞系NP-69中的表达。对聚合酶链反应（PCR）产物进行直接测序，以分析来自上述细胞系的C-KIT序列。CCK-8法检测伊马替尼对细胞增殖的抑制作用。采用Pearson积差相关和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的相关性。结果CNE-1、CNE-2、Hone-1、C-666、SUNE-1和5- 8 F细胞株C-KIT表达水平明显高于NPE细胞株NP-69。在CNE-1、CNE-2、Hone-1和NP-69细胞系中发现了IVS 17 + 78 T>C杂合子，在C-666细胞系中发现了IVS 17 + 78 T>C纯合子，在SUNE-1和5- 8 F细胞系中未发现IVS 17 + 78 T> C突变。伊马替尼对CNE-1、CNE-2、Hone-1、C-666、SUNE-1和5- 8 F的增殖具有剂量依赖性抑制作用。结论鼻咽癌细胞中存在C-KIT过表达和内含子突变，伊马替尼对鼻咽癌细胞增殖具有剂量依赖性抑制作用，但C-KIT过表达、C-KIT突变和伊马替尼的抑制作用之间无明显相关性。
BACKGROUND AND OBJECTIVE 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. METHODS 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. RESULTS 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. CONCLUSION 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.