pEgr1-TRAIL重组质粒联合电离辐射对乳腺癌MCF-7细胞死亡受体通路相关基因和蛋白表达的影响

doi:10.13481/j.1671-587x.20160301

吉林大学学报(医学版) ›› 2016, Vol. 42 ›› Issue (03): 419-423.doi: 10.13481/j.1671-587x.20160301

• 基础研究 • 下一篇

pEgr1-TRAIL重组质粒联合电离辐射对乳腺癌MCF-7细胞死亡受体通路相关基因和蛋白表达的影响

胡志宏¹, 赵大力², 谢忠伟², 王梦莹¹, 胡景坤¹, 龚守良³, 刘扬³, 齐亚莉¹

1. 北华大学公共卫生学院流行病学教研室, 吉林吉林 132011;
2. 吉林省吉林(市)出入境检验检疫局, 吉林吉林 132012;
3. 吉林大学公共卫生学院卫生部放射生物学重点实验室, 吉林长春 130021

收稿日期:2016-01-23 出版日期:2016-05-28 发布日期:2016-06-17
通讯作者: 齐亚莉,副教授,硕士研究生导师(Tel:0432-64608343,E-mail:374494617@qq.com) E-mail:374494617@qq.com
作者简介:胡志宏(1966-),女,吉林省吉林市人,副教授,医学硕士,主要从事肿瘤基因放射治疗方面的研究。
基金资助:
国家自然科学基金资助课题(30970681);吉林省教育厅"十二五"科学技术研究项目资助课题(2014-192)

Effects of pEgr1-TRAIL recombinant plasmid combined with ionizing radiation on related gene and protein expressions of death receptor pathway in breast cancer MCF-7 cells

HU Zhihong¹, ZHAO Dali², XIE Zhongwei², WANG Mengying¹, HU Jingkun¹, GONG Shouliang³, LIU Yang³, QI Yali¹

1. Department of Epidemiology, School of Public Health, Beihua University, Jilin 132001, China;
2. Jilin Province (City) Entry and Exit Inspection and Quarantine Bureau, Jilin Province, Jilin 132012, China;
3. Key Laboratory of Radiobiology, Ministry of Health, School of Public Health, Jilin University, Changchun 130021, China

Received:2016-01-23 Online:2016-05-28 Published:2016-06-17

摘要/Abstract

摘要：

目的: 将Egr1-TRAIL重组质粒转染人乳腺癌MCF-7细胞且加X线照射,探讨MCF-7细胞中死亡受体通路相关基因和蛋白表达的变化。方法: 将MCF-7细胞分为对照组、空质粒组、pEgr1-TRAIL质粒组、4.0GyX射线组、空质粒+4.0GyX射线组和pEgr1-TRAIL+4.0GyX射线组。Real-timePCR法检测各组MCF-7细胞中DR4、caspase-9和caspase-6mRNA表达水平;Western blotting法检测各组MCF-7细胞中DR4、caspase-9和caspase-6蛋白相对表达水平。结果: 经4.0GyX射线照射后4h,与对照组比较,各组MCF-7细胞中DR4、caspase-9和caspase-6mRNA表达水平升高(P<0.01),8h达最高值,之后开始降低,到24h恢复到照射前水平;各组MCF-7细胞中DR4、caspase-9和caspase-6mRNA表达由高到低的顺序为pEgr1-TRAIL+4.0GyX射线组> 空质粒+4.0GyX射线组> pEgr1-TRAIL质粒组> 空质粒组> 4.0GyX射线组> 对照组,其中pEgr1-TRAIL+4.0GyX射线组MCF-7细胞中DR4、caspase-9和caspase-6mRNA表达水平明显高于其他各组(P<0.01)。MCF-7细胞中DR4、caspase-9和caspase-6蛋白在照射后6h开始表达,12h达高峰,48h后细胞中DR4、caspase-9和caspase-6蛋白表达水平仍高于6h时蛋白表达水平。与对照组比较,其他各组MCF-7细胞中蛋白相对表达水平由高到低的顺序为pEgr1-TRAIL+4.0GyX射线组> 空质粒+4.0GyX射线组> 4.0GyX射线组> pEgr1-TRAIL质粒组> 空质粒组> 对照组,其中pEgr1-TRAIL+4.0GyX射线组MCF-7细胞中蛋白相对表达水平明显高于其他各组。结论: pEgr1-TRAIL重组质粒联合放疗对MCF-7细胞具有杀伤和诱导凋亡的作用,其效果优于单纯质粒或单纯照射。

关键词: pEgr1-TRAIL重组质粒, 电离辐射, MCF-7细胞, 死亡受体通路

Abstract:

Objective: To transfer the pEgr1-TRAIL recombinant plasmid into human breast cancer MCF-7 cells and to combine with ionizing radiation,and to discuss the changes of related gene and protein expressions of death receptor pathway in breast cancer MCF-7 cells. Methods: The MCF-7 cells were divided into empty plasmid,pEgr1-TRAIL plasmid,4.0 Gy X-ray,empty plasmid+4.0 Gy X-ray, and pEgr1 TRAIL+4.0 Gy X-ray groups.Real-time PCR method was used to detect the expression levels of DR4,caspase-9,and caspase-6 mRNA in the MCF-7 cells in various groups;Western blotting method was used to detect the relative expression levels of DR4,caspase-9,and caspase-6 protein in the MCF-7 cells in various groups. Results: Compared with control group, the mRNA expression levels of DR4,caspase-9 and caspase-6 in MCF-7 cells in the other groups began to rise 4 h after 4.0 Gy X-ray irradiation(P<0.01),and they reached to the highest levels 8 h later,then reduced to the level of pre-irradiation 24 h later.The order of these mRNA expression levels from high to low in various groups was pEgr1-TRAIL+4.0 Gy X-ray group>empty plasmid+4.0 Gy X-ray group >pEgr1-TRAIL plasmid group >empty plasmid group >4.0 Gy X-ray group>control group;the expression levels of RD4,caspase-9,and caspase-6 mRNA in the MCF-7 cells in pEgr1-TRAIL+4.0 Gy X-ray group were higher than those in other groups (P<0.01).The relative expression levels of DR4,caspase-9 and caspase-6 proteins began to rise 6 h after irradiation, and reached the peak 12 h later; the relative expression levels of those protein 48 h later were still higher than those 6 h after irrdiation.The order of these protein expression levels from high to low in various group was pEgr1 TRAIL+4.0 Gy X-ray group >empty plasmid+4.0 Gy X-ray group >4.0 Gy X-ray group >pEgr1-TRAIL plasmid group >empty plasmid group >control group.The relative expression levels of DR4,caspase-9,and caspase-6 proteins in the MCF-7 cells in pEgr1-TRAIL+4.0 Gy X-ray group were higher than those in the other groups. Conclusion: pEgr1-TRAIL recombinant plasmid combined with radiation can kill and induce the apoptosis of MCF-7 cells,and its effect is better than recombinant plasmid or ionizing radiation used alone.

Key words: pEgr1-TRAIL recombinant plasmid, ionizing radiation, MCF-7 cell, death receptor pathway

中图分类号:

R144.1

胡志宏, 赵大力, 谢忠伟, 王梦莹, 胡景坤, 龚守良, 刘扬, 齐亚莉. pEgr1-TRAIL重组质粒联合电离辐射对乳腺癌MCF-7细胞死亡受体通路相关基因和蛋白表达的影响[J]. 吉林大学学报(医学版), 2016, 42(03): 419-423.

HU Zhihong, ZHAO Dali, XIE Zhongwei, WANG Mengying, HU Jingkun, GONG Shouliang, LIU Yang, QI Yali. Effects of pEgr1-TRAIL recombinant plasmid combined with ionizing radiation on related gene and protein expressions of death receptor pathway in breast cancer MCF-7 cells[J]. Journal of Jilin University Medicine Edition, 2016, 42(03): 419-423.

参考文献

[1] Tait SW,Ichim G,Green DR. Die another way-non-apoptotic mechanisms of cell death [J].J Cell Sci,2014,127(Pt 10):2135-2144.

[2] Chen YJ,Chi CW,Su WC,et al.Lapatinib induces autophagic cell death and inhibits growth of human hepatocellular carcinoma [J].Oncotarget,2014,5(13):4845-4854.

[3] Bai L,Wang S.Targeting apoptosis pathways for new cancer therapeutics [J].Annu Rev Med,2014,65:139-155.

[4] Lin Y,Wang X,Jin H.EGFR-TKI resistance in NSCLC patients:mechanisms and strategies [J].Am J Cancer Res,2014,4(5):411-435.

[5] Bae JH,Shim JH,Cho YS.Chemical regulation of signaling pathways to programmed necrosis [J].Arch Pharm Res,2014,37(6):689-697.

[6] Khaider NG,Lane D,Matte I,et al.Targeted ovarian cancer treatment:the TRAILs of resistance [J].Am J Cancer Res,2012,2(1):75-92.

[7] Chunhacha P,Chanvorachote P.Roles of caveolin-1 on anoikis resistance in non small cell lung cancer [J].Int J Physiol Pathophysiol Pharmacol,2012,4(3):149-155.

[8] Vanden Berghe T,Linkermann A,Jouan-Lanhouet S,et al.Regulated necrosis:the expanding network of non-apoptotic cell death pathways [J].Nat Rev Mol Cell Biol,2014,15(2):135-147.

[9] Jonckheere N,Vincent A,van Seuningen I.Of autophagy and in vivo pancreatic carcinogenesis:the p53 status matters [J]. Clin Res Hepatol Gastroenterol,2014,38(4):423-425.

[10] Bitto A,Lerner CA,Nacarelli T,et al.P62/SQSTM1 at the interface of aging,autophagy,and disease [J].Age (Dordr),2014,36(3):9626.

[11] Sanli T,Steinberg GR,Singh G,et al.AMP-activated protein kinase (AMPK) beyond metabolism:a novel genomic stress sensor participating in the DNA damage response pathway [J].Cancer Biol Ther,2014,15(2):156-169.

[12] 关锋,龚平生,王志成,等.肿瘤坏死因子相关凋亡配体在肿瘤放射治疗中的作用[J].中华放射医学与防护杂志,2014,34(3):231-234.

[13] Shiloh R,Bialik S,Kimchi A.The DAPK family:a structure-function analysis [J].Apoptosis,2014,19(2):286-297.

[14] Suman S,Das TP,Reddy R,et al.The pro-apoptotic role of autophagy in breast cancer [J].Br J Cancer,2014,111(2):309-317.

[15] Ettinger DS,Akerley W,Borghaei H,et al.National comprehensive cancer network:Non-small cell lung cancer,version 2.2013 [J].J Natl Compr Canc Netw,2013,11(3):645-653.

[16] Hsu CL,Chen JH,Chen KY,et al.Advanced non-small cell lung cancer in the elderly:The impact of age and comorbidities on treatment modalities and patient prognosis [J].J Geriatr Oncol,2015,6(4):38-45.

[17] 张卫芳,孙燕,张伟杰,等.TM4SF1在人乳腺癌MCF-7和MDA-MB-231中的表达及其对细胞增殖、迁移能力的影响[J].郑州大学学报:医学版,2016,51(2):240-244.

pEgr1-TRAIL重组质粒联合电离辐射对乳腺癌MCF-7细胞死亡受体通路相关基因和蛋白表达的影响

Effects of pEgr1-TRAIL recombinant plasmid combined with ionizing radiation on related gene and protein expressions of death receptor pathway in breast cancer MCF-7 cells

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