The influence of autophagy-related genes about X-Ray on nasopharyngeal carcinoma CNE2 and CNE2/DDP cells
-
摘要: 目的: 研究放疗抵抗与自噬的关系,为使用药物调节自噬水平来提高鼻咽癌放疗敏感性提供理论基础。方法: 采用流式细胞术分析CNE2及CNE2/DDP细胞照射前后的细胞周期分布;实时荧光定量PCR和免疫荧光染色检测CNE2及CNE2/DDP细胞中自噬特异性基因Beclin 1及微管相关蛋白轻链3β(MAPLC3β)的表达水平。结果: 射线照射后CNE2及CNE2/DDP细胞的G2-M期增多,与放射剂量呈正相关(P<0.05),CNE2/DDP细胞的G2-M期阻滞比CNE2明显(P<0.05)。射线照射后CNE2及CNE2/DDP细胞的Beclin1及MAPLC3β的表达水平增高,与放射剂量呈正相关(P<0.05)。CNE2/DDP细胞的Beclin1及MAPLC3β表达水平均低于CNE2细胞,差异有统计学意义(P<0.05)。结论: 自噬性死亡可能是放射治疗后人鼻咽癌CNE2及CNE2/DDP细胞的死亡方式之一,而CNE2/DDP细胞的放射抵抗可能与低水平的自噬有关。Abstract: Objective: To study the relationship between the radiotherapy resistance and autophagy. To provide a theoretiacal basis for drugs that regulate autophagy to improve radiotherapy sensitivity.Method: Flow cytometry (FCM) was performed to analyze the distribution of the cell cycle of CNE2 and CNE2/DDP cells under the action of X radiation. The expression of autopagy-specific gene Beclin1 and microtubule-associated protein light chain 3β (MAPLC3β) in CNE2 and CNE2/DDP cells was determined by real time PCR and Immumofluorescence staining.Result: CNE2/DDP and their parental CNE2 cells produced the G2-M phase arrest under the action of X radiation.With the radiation dose increasing,The cells which in the G2-M phase were more and more (P<0.05). The G2-M phase arrest in CNE2/DDP cells was more obvious than in CNE2 cells (P<0.05). The expression of Beclin1 and MAPLC3β in CNE2 and CNE2/DDP cells increased under the action of X radiation. What's more, the raise was more and more obvious with the increase of the irradiation dose(P<0.05). The expression levels of Beclin1 and MAPLC3β in CNE2/DDP was lower than that in CNE2 cells (P<0.05).Conclusion: Autophagic cell death may be the one manner of death in nasopharyngeal carcinoma CNE2 and CNE2/DDP cells under the action of X radiation. The radiation resistance of CNE2/DDP cells may be related to the low expression of autophagy-related genes.
-
Key words:
- autophagy /
- radiation resistance /
- Beclin 1 /
- MAPLC3β
-
[1] HENG D M,WEE J,FONG K W,et al.Prognostic factors in 677patients in Singapore with nondisseminated nasopharyngeal carcinoma[J].Cancer,1999,86:1912-1920.
[2] ALI H,AL-SARRAF M.Chemotherapy in advanced nasopharyngeal cancer[J].Oncology(Williston Park),2000,14:1223-1230.
[3] 曹素梅,洪明晃,郭翔,等.血浆EB病毒游离DNA检测对监测鼻咽癌患者预后的意义[J].癌症,2003,22(3):302-306.
[4] TEO P,YU P,LEE W Y,et al.Significant prognosticators after primary radiotherapy in 903nondisseminated nasopharyngeal carcinoma evaluatedby computer tomography[J].Int J Radiat Oncol Biol Phys,1996,36:291-304.
[5] HENG,WE D M,FONG J,et al.Prognostic factors in 677 Patients in Singapore with nondisseminated nasopharyngeal careinoma[J].Cancer,1999,86:1912-1920.
[6] NAGASAW A H,KENG P,HARLEY R,et al.Relationship between Y-ray-induced G2/M Delay and cellular Radiosensitivity[J].Original Article,1994,66:373-379.
[7] KLIONSKY D J,EMR S D.Autophagy as a regulated pathway of cellular degradation[J].Science,2000,290:1717-1721.
[8] OHSUMI Y.Molecular dissection of autophagy:two ubiquitin-like systems[J].Nat Rev Mol Cell Biol,2001,2:211-216.
[9] KOUKOURAKIS1 M I,GIATROMANOLAKI2 A,SIVRIDIS E,et al.Beclin 1over- and underexpression in colorectal cancer:distinct patterns relate to prognosis and tumour hypoxia[J].British J Cancer,2010,10:1209-1214.
[10] MIZSHIMA N,OHSUMI Y,YOSHIMORI T.Autophagosome formation in mammalian cells[J].Cell Struct Funct,2002,27:421-429.
[11] LIANG X H,YU J,BROWN K,et al.Beclin 1contains a leucine-rich nuclear export signal that is required for its autophagy and tumorsuppressor function[J].Cancer Res,2001,61:3443-3449.
[12] KUWAHARA Y,OIKAWA T,OCHIAI Y,et al.Enhancement of autophagy is a potential modality for tumors refractory to radiotherapy[J].Cell Death and Disease,2011,2:2041-4889.
[13] AMARAVADI R K,THOMPSON C B.The roles of therapy-induced autophagy and necrosis in cancer treatment[J].Clin Cancer Res,2007,13:7271-7279.
计量
- 文章访问数: 91
- PDF下载数: 50
- 施引文献: 0