X射线诱导人鼻咽鳞状细胞癌CNE1细胞耐药相关通路的研究

doi:10.3760/cma.j.issn.1673-422X.2017.06.001

摘要/Abstract

摘要： 目的探讨经X射线照射后人鼻咽鳞状细胞癌CNE1细胞耐药性的产生与血小板内皮细胞黏附分子1（PECAM1）/磷酯酰肌醇3激酶（PI3K）/蛋白激酶B（AKT）/Bcl2信号通路的关系。方法①对CNE1细胞进行培养，待细胞进入对数生长期后，进行X射线照射，2 Gy/次，隔天照射1次，累积剂量50 Gy，收集照射后获得的CNE1/R细胞。②CNE1/R细胞培养24 h后，分为3组：对照组、实验组一（经1∶600的PECAM1抗体作用24 h）、实验组二（经10 μmol/L的PI3K抑制剂LY294002作用24 h）。③利用四甲基偶氮唑蓝(MTT)法分别检测对照组和两个实验组CNE1/R细胞对顺铂的半抑制浓度（IC50）值。④半定量反转录聚合酶链反应（RTPCR）技术分别检测对照组和实验组一PI3K mRNA和Bcl2 mRNA的表达情况。⑤半定量RTPCR技术分别检测对照组和实验组二AKT mRNA和Bcl2 mRNA的表达情况。结果①对照组、实验组一和实验组二的IC50值分别为（2.99±0.02）μg/ml、（1.50±0.03）μg/ml、（1.60±0.05）μg/ml，3组间比较差异具有统计学意义（F=4 381.263，P=0.000），两实验组的IC50均较对照组明显降低（t=116.885，P=0.000；t=73.006，P=0.000）。②对照组和实验组一的PI3K mRNA的半定量值分别为1.66±0.01和0.90±0.05(t=50.394，P=0.000)；Bcl2 mRNA的半定量值分别为1.66±0.02和0.71±0.05（t=183.165，P=0.000），差异均具有统计学意义。③对照组和实验组二的AKT mRNA的半定量值分别为1.53±0.01和0.72±0.01（t=135.281，P=0.000）；Bcl2 mRNA的半定量值分别为1.66±0.02和0.63±0.02（t=128.814，P=0.000），差异均具有统计学意义。结论X射线照射后人鼻咽鳞状细胞癌CNE1细胞耐药性的产生，可能与PECAM1通过PI3K/AKT途径调节Bcl2的活性有关。

关键词: 鼻咽肿瘤, 抗药性, 肿瘤, 抗原, CD31, X线

Abstract: ObjectiveTo investigate the relationship between multidrug resistance of Xray irradiated human nasopharyngeal squamous carcinoma cell line CNE1 and platelet endothelial cell adhesion molecule1 (PECMA1)/phosphatidylinositol 3kinase (PI3K)/protein kinase B (AKT)/Bcl2 signaling pathway. Methods①The CNE1 cells were cultured in incubator. Exponentially growing CNE1 cells were exposed to 50 Gy ionizing radiation administered in 25 fractions, 2 Gy per fraction, once every two days, and the CNE1/R cells were collected. ②CNE1/R cells were cultured for 24 h, and divided into three groups: the control group, experimental group one (CNE1/R cells were processed through 1∶600 PECAM1 antibody for 24 h) and experimental group two (CNE1/R cells were processed through 10 μmol/L PI3K inhibitor LY294002 for 24 h). ③The half maximal inhibitory concentration (IC50) values of the control group and two experimental groups of CNE1/R cells were detected by methyl thiazolyl tetrazolium (MTT). ④The mRNA expressions of PI3K and Bcl2 in the control group and experimental group one were measured by semiquantitative reverse transcriptionpolymerase chain reaction (RTPCR). ⑤The mRNA expressions of AKT and Bcl2 in the control group and experimental group two were measured by semiquantitative RTPCR. Results①The IC50 of the control group, experimental group one and two were (2.99±0.02)μg/ml, (1.50±0.03)μg/ml and (1.60±0.05)μg/ml, and the difference among the three groups was statiatically signficant （F=4 381.263， P=0.000）. The IC50 of the two experimental groups were obviously lower than that of the control group (t=116.885, P=0.000; t=73.006, P=0.000). ②The semiquantitative values of PI3K in the control group and experimental group one were 1.66±0.01 and 0.90±0.05 (t=50.394, P=0.000), and the semiquantitative values of Bcl2 were 1.66±0.02 and 0.71±0.05 (t=183.165, P=0.000), and the differences were statistically significant. ③The semiquantitative values of AKT of control group and experimental group two were 1.53±0.01 and 0.72±0.01 (t=135.281, P=0.000), and the semiquantitative values of Bcl2 were 1.66±0.02 and 0.63±0.02 (t=128.814, P=0.000), and the differences were statistically significant. ConclusionPECAM1 induced multidrug resistance of human nasopharyngeal squamous cell carcinoma CNE1 cells may be related to the activity of Bcl2 through the signal pathway of PI3K/AKT.

Key words: Nasopharyngeal neoplasms, Drug resistance, neoplasm, Antigens, CD31, Xrays

皇婷,廖和和,刘晓栋,王若雨. X射线诱导人鼻咽鳞状细胞癌CNE1细胞耐药相关通路的研究[J]. 国际肿瘤学杂志, 2017, 44(6): 401-.

Huang Ting, Liao Hehe, Liu Xiaodong, Wang Ruoyu. Study of signal transduction mechanisms of Xray induced multidrug resistance in nasopharyngeal squamous cell carcinoma CNE1 cells[J]. Journal of International Oncology, 2017, 44(6): 401-.

参考文献

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