Relationship Between NF-κB and Epithelial-to-mesenchymal Transition of Radiation-induced Human Esophageal Carcinoma Cells
-
摘要:目的
探讨NF-κB与放射线诱导的人食管癌细胞上皮间质转化(EMT)间的关系。
方法累积照射食管癌Eca109细胞60 Gy,得到Eca109R60细胞,克隆形成实验检测其放射抵抗性,显微镜下观察细胞形态学变化。Real-time PCR和免疫细胞化学检测Eca109和Eca109R60细胞NF-κB、E-cadherin和Vimentin的表达情况。
结果Eca109R60细胞具备了EMT样表型且放射抵抗性更强。Eca109R60细胞中E-cadherin mRNA表达明显下调(P=0.003),Vimentin、NF-κB mRNA表达升高(P=0.004,P=0.004)。相关性分析显示NF-κB与E-cadherin mRNA表达呈负相关,与Vimentin未见有明显相关性,但已显示出正相关的趋势。免疫细胞化学法检测Eca109R60细胞E-cadherin阳性染色程度明显减弱,Vimentin、NF-κB p65阳性染色程度明显增强。
结论Eca109R60细胞放射抗性可能由EMT与NF-κB信号通路相互作用调控。
Abstract:ObjectiveTo explore the relationship between NF-κB and epithelial-to-mesenchymal transition (EMT) of human esophageal carcinoma cells induced by the radiation.
MethodsWe irradiated cumulatively human esophageal carcinoma cells Eca109 with 60Gy to get Eca109R60 cells. The radio-resistance was detected by means of a standard colony-formation assay. Cells morphological change was observed under microscope. Meanwhile,Real-time PCR and immunocytochemistry were used to detect the expression level of NF-κB,E-cadherin and Vimentin in esophageal epithelial cell lines Eca109 and Eca109R60,respectively.
ResultsEca109R60 cells had EMT-like phenotype and stronger irradiation resistance(IR). In Eca109R60 cells,the expression level of E-cadherin mRNA was significantly down-regulated (P=0.003) . There were significant upregulation in the expression levels of Vimentin and NF-κB mRNA (P=0.004,P=0.004) . It showed a negative correlation between NF-κB and E-cadherin by the correlation analysis. There was not an evident correlation between NF-κB and Vimentin,but a trend to negative correlation. From the result of immunocytochemistry staining,a weaker expression of E-cadherin protein while stronger expression of Vimentin and NF-κB p65 protein were found.
ConclusionThe radio-resistance of human esophageal carcinoma cells Eca109R60 may be regulated through the interaction between EMT and NF-κB signaling pathway.
-
Key words:
- Esophageal carcinoma /
- Radio-resistance /
- Radiotherapy /
- NF-κB /
- E-cadherin /
- Vimentin
-
-
表 1 Eca109与Eca109R60放射生物学参数
Table 1 Radiation biology parameter of Eca109 and Eca109R60 cells
-
[1] Stewart BW,Wild C. World Cancer Report 2014[M]. Lyon:The International Agency for Research on Cancer,2014: 529-30.
[1] Stewart BW, Wild C. World Cancer Report 2014[M]. Lyon:The International Agency for Research on Cancer, 2014: 529-30. [2] Theys J, Jutten B, Habets R, et al. E-cadherin loss associated with EMT promotes radioresistance in human tumor cells[J]. Radiother Oncol, 2011, 99(3): 392-7. [2] Theys J,Jutten B,Habets R,et al. E-cadherin loss associated with EMT promotes radioresistance in human tumor cells[J]. Radiother Oncol,2011,99(3): 392-7. doi: 10.1016/j.radonc.2011.05.044
[3] Lan F,Yue X,Ren G,et al. miR-15a/16 enhances radiation sensitivity of non-small cell lung cancer cells by targeting the TLR1/NF-κB signaling pathway[J]. Int J Radiat Oncol Biol Phys,2015,91(1): 73-81. doi: 10.1016/j.ijrobp.2014.09.021
[3] Lan F, Yue X, Ren G, et al. miR-15a/16 enhances radiation sensitivity of non-small cell lung cancer cells by targeting the TLR1/NF-κB signaling pathway[J]. Int J Radiat Oncol Biol Phys, 2015, 91(1): 73-81. [4] Zhang Y, Wei Y, Zhu Z, et al. Icariin enhances radiosensitivity of colorectal cancer cells by suppressing NF-κB activity[J]. Cell Biochem Biophys, 2014, 69(2): 303-10. [4] Zhang Y,Wei Y,Zhu Z,et al. Icariin enhances radiosensitivity of colorectal cancer cells by suppressing NF-κB activity[J]. Cell Biochem Biophys,2014,69(2): 303-10. doi: 10.1007/s12013-013-9799-x
[5] Ahmed KM, Zhang H, Park CC. NF-κB regulates radioresistance mediated by β1-integrin in three-dimensional culture of breast cancer cells[J]. Cancer Res, 2013, 73(12): 3737-48. [5] Ahmed KM,Zhang H,Park CC. NF-κB regulates radioresistance mediated by β1-integrin in three-dimensional culture of breast cancer cells[J]. Cancer Res,2013,73(12): 3737-48. doi: 10.1158/0008-5472.CAN-12-3537
[6] 阴骏,李涛. 食管癌个体化综合治疗的研究[J]. 国际肿瘤学杂志,2012,39(7): 526-9. Yin J,Li T. Individual comprehensive therapy study for esophageal carinoma[J]. Guo Ji Zhong Liu Xue Za Zhi,2012,39(7): 526-9.in J,Li T. Individual comprehensive therapy study for esophageal carinoma[J]. Guo Ji Zhong Liu Xue Za Zhi,2012,39(7): 526-9.
[6] 阴骏, 李涛. 食管癌个体化综合治疗的研究[J]. 国际肿瘤学杂 志, 2012, 39(7): 526-9. [Yin J, Li T. Individual comprehensive therapy study for esophageal carinoma[J]. Guo Ji Zhong Liu Xue Za Zhi, 2012, 39(7): 526-9.] [7] Che SM,Zhang XZ,Liu XL,et al. The radiosensitization effect of NS398 on esophageal cancer stem cell-like radioresistant cells[J]. Dis Esophagus,2011,24(4): 265-73. doi: 10.1111/j.1442-2050.2010.01138.x
[7] Che SM, Zhang XZ, Liu XL, et al. The radiosensitization effect of NS398 on esophageal cancer stem cell-like radioresistant cells[J]. Dis Esophagus, 2011, 24(4): 265-73. [8] 李文雯, 张琳丽, 胡国清. 转录因子Snail对食管癌Eca-109细胞 侵袭和迁移的影响[J]. 肿瘤防治研究, 2015, 42(4): 319-23. [Li WW, Zhang LL, Hu GQ. Effect of transcription factor snail on invasion and migration of esophageal carcinoma cell line Eca- 109[J]. Zhong Liu Fang Zhi Yan Jiu, 2015, 42(4): 319-23.] [8] 李文雯,张琳丽,胡国清. 转录因子Snail对食管癌Eca-109细胞侵袭和迁移的影响[J]. 肿瘤防治研究,2015,42(4): 319-23. http://www.zlfzyj.com/CN/abstract/abstract8457.shtml Li WW,Zhang LL,Hu GQ. Effect of transcription factor snail on invasion and migration of esophageal carcinoma cell line Eca-109[J]. Zhong Liu Fang Zhi Yan Jiu,2015,42(4): 319-23.i WW,Zhang LL,Hu GQ. Effect of transcription factor snail on invasion and migration of esophageal carcinoma cell line Eca-109[J]. Zhong Liu Fang Zhi Yan Jiu,2015,42(4): 319-23. http://www.zlfzyj.com/CN/abstract/abstract8457.shtml
[9] Tsukamoto H,Shibata K,Kajiyama H,et al. Irradiation-induced epithelial-mesenchymal transition (EMT) related to invasive potential in endometrial carcinoma cells[J]. Gynecol Oncol,2007,107(3): 500-4. doi: 10.1016/j.ygyno.2007.08.058
[9] Tsukamoto H, Shibata K, Kajiyama H, et al. Irradiation-induced epithelial-mesenchymal transition (EMT) related to invasive potential in endometrial carcinoma cells[J]. Gynecol Oncol, 2007, 107(3): 500-4. [10] Su H, Jin X, Shen L, et al. Inhibition of cyclin D1 enhances sensitivity to radiotherapy and reverses epithelial to mesenchymal transition for esophageal cancer cells[J]. Tumour Biol, 2016, 37(4): 5355-63. [10] Su H,Jin X,Shen L,et al. Inhibition of cyclin D1 enhances sensitivity to radiotherapy and reverses epithelial to mesenchymal transition for esophageal cancer cells[J]. Tumour Biol,2016,37(4): 5355-63. doi: 10.1007/s13277-015-4393-z
[11] Meyn RE,Milas L,Ang KK. The role of apoptosis in radiation oncology[J]. Int J Radiat Biol,2009,85(2): 107-15. doi: 10.1080/09553000802662595
[11] Meyn RE, Milas L, Ang KK. The role of apoptosis in radiation oncology[J]. Int J Radiat Biol, 2009, 85(2): 107-15. [12] Bhat KP, Balasubramaniyan V, Vaillant B, et al. Mesenchymal differentiation mediated by NF-κB promotes radiation resistance in glioblastoma[J]. Cancer Cell, 2013, 24(3): 331-46. [12] Bhat KP,Balasubramaniyan V,Vaillant B,et al. Mesenchymal differentiation mediated by NF-κB promotes radiation resistance in glioblastoma[J]. Cancer Cell,2013,24(3): 331-46. doi: 10.1016/j.ccr.2013.08.001
[13] Xue XY,Liu ZH,Jing FM,et al. Relationship between NRAGE and the radioresistance of esophageal carcinoma cell line TE13R120[J]. Chin J Cancer,2010,29(10): 900-6. doi: 10.5732/cjc.010.10141
[13] Xue XY, Liu ZH, Jing FM, et al. Relationship between NRAGE and the radioresistance of esophageal carcinoma cell line TE13R120[J]. Chin J Cancer, 2010, 29(10): 900-6. [14] Duru N,Candas D,Jiang G,et al. Breast cancer adaptive resistance: HER2 and cancer stem cell repopulation in a heterogeneous tumor society[J]. J Cancer Res Clin Oncol,2014,140(1):1-14. doi: 10.1007/s00432-013-1494-1
[14] Duru N, Candas D, Jiang G, et al. Breast cancer adaptive resistance: HER2 and cancer stem cell repopulation in a heterogeneous tumor society[J]. J Cancer Res Clin Oncol, 2014, 140(1):1-14. [15] Sinha S, Ghildiyal R, Mehta VS, et al. ATM-NFκB axis-driven TIGAR regulates sensitivity of glioma cells to radiomimetics in the presence of TNFα[J]. Cell Death Dis, 2013, 4: e615. [15] Sinha S,Ghildiyal R,Mehta VS,et al. ATM-NFκB axis-driven TIGAR regulates sensitivity of glioma cells to radiomimetics in the presence of TNFα[J]. Cell Death Dis,2013,4: e615. doi: 10.1038/cddis.2013.128
[16] Sun Q, Yao X, Ning Y, et al. Overexpression of response gene to complement 32(RGC32) promotes cell invasion and induces epithelial-mesenchymal transition in lung cancer cells via the NF- κB signaling pathway[J]. Tumour Biol, 2013, 34(5): 2995-3002. [16] Sun Q,Yao X,Ning Y,et al. Overexpression of response gene to complement 32(RGC32) promotes cell invasion and induces epithelial-mesenchymal transition in lung cancer cells via the NF-κB signaling pathway[J]. Tumour Biol,2013,34(5): 2995-3002. doi: 10.1007/s13277-013-0864-2
[17] Huber MA, Azoitei N, Baumann B, et al. NF-kappaB is essential for epithelial-mesenchymal transition and metastasis in a model of breast cancer progression[J]. J Clin Invest, 2004, 114(4): 569-81. [17] Huber MA,Azoitei N,Baumann B,et al. NF-kappaB is essential for epithelial-mesenchymal transition and metastasis in a model of breast cancer progression[J]. J Clin Invest,2004,114(4): 569-81. doi: 10.1172/JCI200421358
[18] 石磊,王世明,贺杰峰. 核转录因子-κB在胰腺癌中的表达及其与上皮细胞间质转化的关系[J]. 肿瘤研究与临床,2011,23(3): 191-3. http://cdmd.cnki.com.cn/Article/CDMD-10114-1011092173.htm Shi L,Wang SM,He JF. Expression of NF-κB and its relation with epithelial-mesenchymal transition in pancreatic cancer[J]. Zhong Liu Yan Jiu Yu Lin Chuang,2011,23(3): 191-3.hi L,Wang SM,He JF. Expression of NF-κB and its relation with epithelial-mesenchymal transition in pancreatic cancer[J]. Zhong Liu Yan Jiu Yu Lin Chuang,2011,23(3): 191-3. http://cdmd.cnki.com.cn/Article/CDMD-10114-1011092173.htm
[18] 石磊, 王世明, 贺杰峰. 核转录因子-Κ B在胰腺癌中的表达 及其与上皮细胞间质转化的关系[J]. 肿瘤研究与临床, 2011, 23(3): 191-3. [Shi L, Wang SM, He JF. Expression of NF-κB and its relation with epithelial-mesenchymal transition in pancreatic cancer[J]. Zhong Liu Yan Jiu Yu Lin Chuang, 2011, 23(3): 191-3.] [19] Yan Z,Yin H,Wang R,et al. Overexpression of integrin-linked kinase (ILK) promotes migration and invasion of colorectal cancer cells by inducing epithelial-mesenchymal transition via NF-κB signaling[J]. Acta Histochem,2014,116(3): 527-33. doi: 10.1016/j.acthis.2013.11.001
[19] Yan Z, Yin H, Wang R, et al. Overexpression of integrin-linked kinase (ILK) promotes migration and invasion of colorectal cancer cells by inducing epithelial-mesenchymal transition via NF-κB signaling[J]. Acta Histochem, 2014, 116(3): 527-33.