普奈洛尔对胃癌放疗敏感性影响及其机制的实验研究
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摘要
目的:从体内实验探究普萘洛尔对胃癌放疗敏感性的作用。方法:使用胃癌细胞株SGC-7901和BALA/C-nunu裸鼠皮下移植瘤模型,利用β受体阻滞剂干预裸鼠,通过测量胃癌移植瘤重量和体积判断肿瘤生长情况,通过免疫组织化学法和Western blot法研究NF-κB、EGFR、COX-2和VEGF的表达。结果:在裸鼠体内实验发现,在放疗前给予普奈洛尔可以明显减小移植瘤的体积及重量,起到抑制肿瘤生长和促进肿瘤凋亡的作用。通过免疫组织化学染色和Western blot发现放疗前使用普奈洛尔较单独使用放射治疗,可以降低细胞及组织NF-κB表达,并下调其下游相关分子VEGF、EGFR和COX-2的表达,从而抑制胃癌细胞的增殖及抑制其侵袭转移的能力。而在单独使用相同剂量普奈洛尔进行干预未见上述变化。结论:通过β受体阻滞剂普奈洛尔可以加强细胞和组织对放射治疗的敏感性,从而阻断NF-κB表达,下调核转录因子下游相关分子VEGF、EGFR和COX-2的表达,抑制胃癌细胞的增殖及抑制其侵袭转移的能力。提示β受体阻滞剂可作为胃癌的放疗增敏剂,有抗增殖、促凋亡、抑制侵袭和转移能力,为提高胃癌放疗效果提供基础,为探索新的放疗增敏剂提供理论依据。
Objective: To evaluate if propranolol could enhance radiosensitivity and explore its precise molecular mechanism in gastric cancer in vivo study. Methods: Human gastric adenocarcinoma cell lines( SGC-7901) were cultured for research. Tumor models were created by nude mice and then treated with propranolol or/and radiation.Then,the data of tumor weight and volume were obtained for the statistical analysis. Moreover,the expression of NF-κB,EGFR,COX-2 and VEGF were detected by Western blot and immunohistochemical stain. Results: In nude mice,it was found that giving propranolol before radiotherapy can significantly reduce the volume and weight of transplanted tumor,inhibit tumor growth and promote tumor apoptosis. Immunohistochemical staining and Western blot found that the use of propranolol before radiotherapy compared with radiotherapy can reduce the cell and tissue NF-κB expression and down-regulate expression of VEGF,EGFR and COX-2 genes,thereby inhibiting gastric cancer cell proliferation and inhibiting the ability of invasion and metastasis. Conclusion: These results suggested that propranolol enhanced the sensitivity of gastric cancer to radiation through the inhibition of β-ARs and the downstream NF-κB-VEGF/EGFR/COX-2 pathway.
Objective: To evaluate if propranolol could enhance radiosensitivity and explore its precise molecular mechanism in gastric cancer in vivo study. Methods: Human gastric adenocarcinoma cell lines( SGC-7901) were cultured for research. Tumor models were created by nude mice and then treated with propranolol or/and radiation.Then,the data of tumor weight and volume were obtained for the statistical analysis. Moreover,the expression of NF-κB,EGFR,COX-2 and VEGF were detected by Western blot and immunohistochemical stain. Results: In nude mice,it was found that giving propranolol before radiotherapy can significantly reduce the volume and weight of transplanted tumor,inhibit tumor growth and promote tumor apoptosis. Immunohistochemical staining and Western blot found that the use of propranolol before radiotherapy compared with radiotherapy can reduce the cell and tissue NF-κB expression and down-regulate expression of VEGF,EGFR and COX-2 genes,thereby inhibiting gastric cancer cell proliferation and inhibiting the ability of invasion and metastasis. Conclusion: These results suggested that propranolol enhanced the sensitivity of gastric cancer to radiation through the inhibition of β-ARs and the downstream NF-κB-VEGF/EGFR/COX-2 pathway.
引文
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[14]Lin J,Wu H,Shi H,et al. Combined inhibition of epidermal growth factor receptor and cyclooxygenase-2 leads to greater anti-tumor activity of docetaxel in advanced prostate cancer[J].PLo S One,2013,8(10):e76169.
[15]Pang LY,Hurst EA,Argyle DJ. Cyclooxygenase-2:A role in cancer stem cell survival and repopulation of cancer cells during therapy[J]. Stem Cells Int,2016,2016:2048731.
[16]Yu T,Lao X,Zheng H. Influencing COX-2 activity by COX related pathways in inflammation and cancer[J]. Mini Rev Med Chem,2016,16(15):1230-1243.
[17]Salehifar E,Hosseinimehr SJ. The use of cyclooxygenase-2 inhibitors for improvement of efficacy of radiotherapy in cancers[J].Drug Discov Today,2016,21(4):654-662.
[18]Zahra S,Seyed JH. Synergistic effect of epidermal growth factor receptor inhibitors and ionization radiation in cancer treatment[J].Recent Patents on Anti-Cancer Drug Discovery,2007,12(4):1-17.
[19]Komposch K,Sibilia M. EGFR signaling in liver diseases[J]. Int J Mol Sci,2015,17(1):E30.
[20]Cuneo KC,Nyati MK,Ray D,et al. EGFR targeted therapies and radiation:Optimizing efficacy by appropriate drug scheduling and patient selection[J]. Pharmacol Ther,2015,10(154):67-77.
[21]Li C,Huang S,Armstrong EA,et al. Antitumor effects of MEHD7945A,a dual-specific antibody against EGFR and HER3,in combination with radiation in lung and head and neck cancers[J]. Mol Cancer Ther,2015,14(9):2049-2059.
[22]Jiang X,Engelbach JA,Yuan L,et al. Anti-VEGF antibodies mitigate the development of radiation necrosis in mouse brain[J].Clin Cancer Res,2014,20(10):2695-2702.
[23]Gao X,Zhao Y,Stemmer-Rachamimov AO,et al. Anti-VEGF treatment improves neurological function and augments radiation response in NF2 schwannoma model[J]. Proc Natl Acad Sci USA,2015,112(47):14676-14681.
[24]Matsumoto K,Ema M. Roles of VEGF-A signalling in development,regeneration,and tumours[J]. J Biochem,2014,156(1):1
[25]Sclabas GM,Uwagawa T,Schmidt C,et al. Nuclear factorκB activation is a potential target for preventing pancreatic carcinoma by aspirin[J]. Cancer,2005,103(12):2485-2490.
[26]Takada Y,Kobayashi Y,Aggarwal BB. Evodiamine abolishes constitutive and inducible NF-κB activation by inhibiting In Ba kinase activation,thereby suppressing NF-κB-regulated antiapoptotic and metastatic gene expression,up-regulating apoptosis,and inhibiting invasion[J]. J Biol Chem,2005,280(17):17203-17212.
[27]Takada Y,Murakami A,Aggarwal BB. Zerumbone abolishes NF-κB and In Ba kinase activation leading to suppression of antiapoptotic and metastatic gene expression,upregulation of apoptosis,and downregulation of invasion[J]. Oncogene,2005,24(46):6957-6969.
[28]Shin VY,Jin HC,Ng EK,et al. Nicotine and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone induce cyclooxygenase-2 activity in human gastric cancer cells:Involvement of nicotinic acetylcholine receptor(nAChR)and beta-adrenergic receptor signaling pathways[J]. Toxicol Appl Pharmacol,2008,233(2):254-261.
[29]Huang RY,Chen GG. Cigarette smoking,cyclooxygenase-2 pathway and cancer[J]. Biochim Biophys Acta,2011,1815(2):158-169.
[30]Wang W,Chin-Sheng H,Kuo LJ,et al. NNK enhances cell migration throughα7-nicotinic acetylcholine receptor accompanied by increased of fibronectin expression in gastric cancer[J]. Ann Surg Oncol,2012,19(S3):S580-588.
[2]Tatsuta M,Iishi H,Yamamura H,et al. Enhancement by propranolol of the inhibitory effect of tetragastrin on gastric carcinogenesis induced by N-methyl-N'-nitro-N-nitrosoguanidine in Wistar rats[J]. Cancer Res,1987,47(1):111-114.
[3]Iishi H,Tatsuta M,Baba M,et al. Promotion by the alpha-adrenoceptor agonist phenylephrine,but not by the beta-adrenoceptor agonist isoproterenol,of gastric carcinogenesis induced by N-methyl-N'-nitro-N-nitrosoguanidine in Wistar rats[J]. Cancer Lett,1998,122(1-2):61-65.
[4]Luna SL,Neuman S,Aguilera J,et al. In vivoβ-adrenergic blockade by propranolol prevents isoproterenol-induced polycystic ovary in adult rats[J]. Horm Metab Res,2012,44(9):676-681.
[5]Takahashi K,Kaira K,Shimizu A,et al. Clinical significance ofβ2-adrenergic receptor expression in patients with surgically resected gastric adenocarcinoma[J]. Tumour Biol,2016,37(10):13885-13892.
[6]Chang PY,Huang WY,Lin CL,et al. Propranolol reduces cancer risk:A population-based cohort study[J]. Medicine(Baltimore),2015,94(27):e1097.
[7]Lu YJ,Geng ZJ,Sun XY,et al. Isoprenaline induces epithelialmesenchymal transition in gastric cancer cells[J]. Mol Cell Biochem,2015,408(1-2):1-13.
[8]Wei B,Sun X,Geng Z,et al. Isoproterenol regulates CD44 expression in gastric cancer cells through STAT3/microRNA373 cascade[J]. Biomaterials,2016,10(105):89-101.
[9]Kalita B,Ranjan R,Singh A,et al. A combination of podophyllotoxin and rutin attenuates radiation induced gastrointestinal injury by negatively regulating nf-κb/p53 signaling in lethally irradiated mice[J]. PLo S One,2016,11(12):e0168525.
[10]Pordanjani SM,Hosseinimehr SJ. The role of NF-κB inhibitors in cell response to radiation[J]. Curr Med Chem,2016,23(34):3951-3963.
[11]Zhang W,Kang M,Zhang T,et al. Triptolide combined with radiotherapy for the treatment of nasopharyngeal carcinoma via NF-κB-related mechanism[J]. Int J Mol Sci,2016,17(12):E2139.
[12]Park M,Yoon HJ,Kang MC,et al. PTK7 regulates radioresistance through nuclear factor-kappa B in esophageal squamous cell carcinoma[J]. Tumour Biol,2016,37(10):14217-14224.
[13]Adams J. The proteasome,structure,function,and role in the cell[J]. Cancer Treat Rev,2003,29(s1):3-9.
[14]Lin J,Wu H,Shi H,et al. Combined inhibition of epidermal growth factor receptor and cyclooxygenase-2 leads to greater anti-tumor activity of docetaxel in advanced prostate cancer[J].PLo S One,2013,8(10):e76169.
[15]Pang LY,Hurst EA,Argyle DJ. Cyclooxygenase-2:A role in cancer stem cell survival and repopulation of cancer cells during therapy[J]. Stem Cells Int,2016,2016:2048731.
[16]Yu T,Lao X,Zheng H. Influencing COX-2 activity by COX related pathways in inflammation and cancer[J]. Mini Rev Med Chem,2016,16(15):1230-1243.
[17]Salehifar E,Hosseinimehr SJ. The use of cyclooxygenase-2 inhibitors for improvement of efficacy of radiotherapy in cancers[J].Drug Discov Today,2016,21(4):654-662.
[18]Zahra S,Seyed JH. Synergistic effect of epidermal growth factor receptor inhibitors and ionization radiation in cancer treatment[J].Recent Patents on Anti-Cancer Drug Discovery,2007,12(4):1-17.
[19]Komposch K,Sibilia M. EGFR signaling in liver diseases[J]. Int J Mol Sci,2015,17(1):E30.
[20]Cuneo KC,Nyati MK,Ray D,et al. EGFR targeted therapies and radiation:Optimizing efficacy by appropriate drug scheduling and patient selection[J]. Pharmacol Ther,2015,10(154):67-77.
[21]Li C,Huang S,Armstrong EA,et al. Antitumor effects of MEHD7945A,a dual-specific antibody against EGFR and HER3,in combination with radiation in lung and head and neck cancers[J]. Mol Cancer Ther,2015,14(9):2049-2059.
[22]Jiang X,Engelbach JA,Yuan L,et al. Anti-VEGF antibodies mitigate the development of radiation necrosis in mouse brain[J].Clin Cancer Res,2014,20(10):2695-2702.
[23]Gao X,Zhao Y,Stemmer-Rachamimov AO,et al. Anti-VEGF treatment improves neurological function and augments radiation response in NF2 schwannoma model[J]. Proc Natl Acad Sci USA,2015,112(47):14676-14681.
[24]Matsumoto K,Ema M. Roles of VEGF-A signalling in development,regeneration,and tumours[J]. J Biochem,2014,156(1):1
[25]Sclabas GM,Uwagawa T,Schmidt C,et al. Nuclear factorκB activation is a potential target for preventing pancreatic carcinoma by aspirin[J]. Cancer,2005,103(12):2485-2490.
[26]Takada Y,Kobayashi Y,Aggarwal BB. Evodiamine abolishes constitutive and inducible NF-κB activation by inhibiting In Ba kinase activation,thereby suppressing NF-κB-regulated antiapoptotic and metastatic gene expression,up-regulating apoptosis,and inhibiting invasion[J]. J Biol Chem,2005,280(17):17203-17212.
[27]Takada Y,Murakami A,Aggarwal BB. Zerumbone abolishes NF-κB and In Ba kinase activation leading to suppression of antiapoptotic and metastatic gene expression,upregulation of apoptosis,and downregulation of invasion[J]. Oncogene,2005,24(46):6957-6969.
[28]Shin VY,Jin HC,Ng EK,et al. Nicotine and 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanone induce cyclooxygenase-2 activity in human gastric cancer cells:Involvement of nicotinic acetylcholine receptor(nAChR)and beta-adrenergic receptor signaling pathways[J]. Toxicol Appl Pharmacol,2008,233(2):254-261.
[29]Huang RY,Chen GG. Cigarette smoking,cyclooxygenase-2 pathway and cancer[J]. Biochim Biophys Acta,2011,1815(2):158-169.
[30]Wang W,Chin-Sheng H,Kuo LJ,et al. NNK enhances cell migration throughα7-nicotinic acetylcholine receptor accompanied by increased of fibronectin expression in gastric cancer[J]. Ann Surg Oncol,2012,19(S3):S580-588.