Notch3在鼻咽癌中的基础研究及临床意义

目的：探讨Notch3是否可以作为判断鼻咽癌复发、转移和化疗预后的分子生物学指标。方法 West-ern blot检测鼻咽癌细胞系CNE2沉默Notch3的效果，球形成实验分析沉默Notch3对癌症干细胞的影响；通过Western blot检测上皮细胞标志E-cadherin和间质细胞标志Vimentin的表达分析沉默Notch3对鼻咽癌细胞EMT的影响；检测鼻咽癌细胞上清中乳酸的浓度分析在鼻咽癌中沉默Notch3对糖酵解的影响；Western blot检测顺铂作用后Notch3的表达，判断Notch3是否可以作为鼻咽癌对化疗药物敏感的指标。结果Western blot结果显示寡核苷酸干扰序列可以明显抑制沉默鼻咽癌细胞系CNE2中Notch3的表达。通过球形成实验发现在鼻咽癌细胞系CNE2中沉默Notch3后球形结构明显增加，说明沉默Notch3后干细胞增加。通过Western blot发现鼻咽癌细胞系CNE2沉默Notch3后上皮细胞标志分子E-cadherin表达减少，间质细胞标志分子Vimentin表达升高，说明沉默促进鼻咽癌细胞EMT转化。通过检测上清液中乳酸的浓度发现鼻咽癌细胞CNE2发现沉默Notch3后乳酸浓度升高，表明糖酵解增加。 Western blot发现化疗药物顺铂作用鼻咽癌细胞系CNE2后Notch3表达减少，表明Notch3可以作为判断化疗药物有效的指标。结论在鼻咽癌细胞系CNE2中沉默Notch3后癌症干细胞增加，促进EMT转换，糖酵解增加。化疗药物顺铂作用CNE2细胞后Notch3表达减少。因此，Notch3可以作为判断鼻咽癌复发、转移和化疗预后的分子生物学指标。

The significance of Notch3 in nasopharyngeal carcinoma

Objective To explore the possibility of Notch 3 as a molecular biological index to evaluate the recurrence , metastasis and prognosis of nasopharyngeal cancer .Methods Western Blot was used to detect the effects of silencing Notch 3 in nasopharyngeal carcinoma cell lines-CNE2 by small interfering RNA ( siRNA) .The sphere forming assay was performed to analyze the influence of silencing Notch 3 in CNE2 on the number of cancer stem cells .The expression levels of E-cadherin and VImentin were detected by Western Blot to observe the influence of silencing Notch 3 in CNE2 on EMT.The effects of silencing Notch3 in CNE2 on glycolysis were observed by detecting the concentration of lactic acid .The expression levels of Notch 3 were determine after CNE2 cells were treated by cisplatin in order to evaluate whether Noth 3 could be regarded as an index of chemotherapy sensitivity.Results The results by Western Blot showed that the siRNA could effectively silence the expressions of Notch3 in CNE2 cells.The results by sphere forming assay showed that the sphericity structure was obviously increased after silencing Notch3 in CNE2 cells, which suggested that silencing Notch 3 increased the number of cancer stem cells .After silencing Notch3 in CNE2 cells the expression levels of epithelial marker -E-cadherin were decreased , however , the expression levels of mesenchymal cell marker molecule -Vimentin were increased , which indicated that silencing Notch 3 promoted EMT transformation of CNE2 cells.Moreover lactate concentration was increased in CNE 2 cells after silencing Notch3,which suggested that silencing Notch 3 in CNE2 cells could increase glycolysis .Western Blot results showed that the expression levels of Notch3 were decreased after CNE2 cells were interfered by cisplatin,which indicated that Notch3 could be regarded as an index to evaluate the efficiency of chemotherapeutics .Conclusion In nasopharyngeal carcinoma cell lines-CNE2 cells, silencing Notch3 can increase the number of cancer stem cell , promote EMT transition and enhance glycolysis . The expression levels of Notch3 are decreased after CNE2 cells are interfered by cisplatin.Thus Notch3 can be regarded as an index to evaluate the recurrence , metastasis and prognosis of nasopharyngeal cancer .