叔丁基双氧水诱导HEI-OC1细胞株凋亡与STATs mRNA表达的相关性分析

Analysis of the Relevance between t-BHP Induced Apoptosis and STATs mRNA Expression in HEI-OC1 Cells

曾梓豪;罗璇;姚裕恒;王军义;

1:广东药科大学公共卫生学院

摘要
目的通过分析氧化应激损伤的凋亡HEI-OC1细胞各信号转导及转录激活子(signal transducers and activators of transcription,STATs)的表达,探讨酪氨酸激酶-信号转导及转录激活子(janus kinase-signal transducers and activators of transcription,JAK-STATs)信号通路在毛细胞凋亡中的生物学作用。方法采用不同浓度(0、20、40、60 μM)叔丁基过氧化氢(tert-butyl hydroperoxide,t-BHP)染毒构建HEI-OC1细胞株凋亡模型(0μM浓度为对照组),Annexin V-FITC/PI法检测20、40、60 μM浓度t-BHP染毒组细胞凋亡水平,实时荧光定量PCR检测STATs mRNA表达水平。结果对照组及20、40、60 μM t-BHP染毒组的细胞凋亡率分别为3.9%、7.4%、32.0%、91.2%;与对照组相比,各浓度T-BHP染毒组STAT1mRNA、STAT3mRNA、STAT5mRNA、STAT6 mRNA的表达水平均显著性下降(分别为F=5 534.302,P<0.01;F=146.038,P<0.01;F=685.929,P<0.01;F=516.11,P<0.01),各浓度t-BHP染毒组间差异无统计学意义(P>0.05);与对照组相比各浓度t-BHP染毒组STAT2mRNA的表达水平均有显著变化(F=1 259.148,P<0.01),20 μM染毒组STAT2 mRNA的表达水平明显下降(P<0.01),但40、60 μM染毒组均明显升高(P<0.01);STAT4 mRNA在HEI-OC1细胞株中未见表达。结论 JAK-STAT2信号通路具有促进毛细胞氧化应急损伤凋亡的生物学作用,JAK-STAT1/STAT3/STAT5/STAT6信号通路有抑制毛细胞氧化应急损伤凋亡的生物学作用。
关键词
氧化应激;HEI-OC1细胞株;凋亡;JAK-STAT信号通路;信号传导及转录激活因子
基金项目(Foundation):
广东省科技计划项目(2014A0202126)
作者
曾梓豪;罗璇;姚裕恒;王军义;
参考文献

1 Gul F,Muderris T,Yalciner G,et al.A comprehensive study of oxidative stress in sudden hearing loss[J].Eur Arch Otorhinolaryngol,2017,274:1301.

2 Song EA.Docosahexaenoic acid induces oxidative DNA damage and apoptosis,and enhances the chemosensitivity of cancer cells[J].International Journal of Molecular Sciences,2016,17:E1257.

3 Motaghinejad M,Motevalian M,Babalouei F,et al.Possible involvement of CREB/BDNF signaling pathway in neuroprotective effects of topiramate against methylphenidate induced apoptosis,oxidative stress and inflammation in isolated hippocampus of rats:molecular,biochemical and histological evidences[J].Brain Res Bull,2017,132:82.

4 Dalg9CT,Kaymaz BT,zkan MC,et al.Investigating the role of JAK/STAT pathway on dasatinib-induced apoptosis for CML cell model K562[J].Clin Lymphoma Myeloma Leuk,2015,15(Suppl):S161.

5 Groner B.Jak Stat signaling and cancer:opportunities,benefits and side effects of targeted inhibition[J].Mol Cell Endocrinol,2017,451:1.

6 Meier JA.Toward a new STATe:the role of STATs in mitochondrial function[J].Semin Immunol,2014,26:20.

7 Zundler S.Integrating immunologic signaling networks:the JAK/STAT pathway in colitis and colitis-associated cancer[J].Vaccines,2016,4:E5.

8 Al Zaid Siddiquee K.STAT3as a target for inducing apoptosis in solid and hematological tumors[J].Cell Research,2008,18:254.

9 Liszewski W,Naym DG,Biskup E.Psoralen with ultraviolet A-induced apoptosis of cutaneous lymphoma cell lines is augmented by type I interferons via the JAK1-STAT1pathway[J].Photodermatol Photoimmunol Photomed,2017,33:164.

10 Arzt L,Halbwedl I,Gogg-Kamerer M.Signal transducer and activator of transcription 1(STAT1)knock-down induces apoptosis in malignant pleural mesothelioma[J].Pathology Oncology Research,2017,23:595.

11 Chowdhury FZ.STAT2:a shape-shifting anti-viral super STAT[J].JJAK-STAT,2013,2:e23633.

12 Shodeinde A,Ginjupalli K,Lewis HD,et al.STAT3inhibition induces apoptosis in cancer cells independent of STAT1or STAT2[J].Journal of Molecular Biochemistry,2013,2:18.

13 Scarzello AJ,Romero-Weaver AL,Maher SG,et a1.A mutation in the SH2 domain of STAT2 prolongs tyrosine phosphorylation of STAT1and promotes type I IFN-induced apoptosis[J].Mol Biol Cell,2007,18:2455.

14 Romero-Weaver AL,Wang HW,Steen HC,et al.Resistance to IFN-alpha-induced apoptosis is linked to a loss of STAT2[J].Molecular Cancer Research,2010,8:80.

15 Al Zaid Siddiquee K.STAT3as a target for inducing apoptosis in solid and hematological tumors[J].Cell Research,2008,18:254.

16 Geiger JL,Grandis JR.The STAT3pathway as a therapeutic target in head and neck cancer:barriers and innovations[J].Oral Oncol,2016,56:84.

17 Maddur MS,Miossec P,Kaveri SV.Th17cells:biology,pathogenesis of autoimmune and inflammatory diseases,and therapeutic strategies[J].The American Journal of Pathology,2012,181:8.

18 Varikuti S,Oghumu S,Natarajan G,et al.STAT4is required for the generation of Th1and Th2,but not Th17immune responses during monophosphoryl lipid A adjuvant activity[J].International Immunology,2016,28:565.

19 Mütze J,Roth J,Gerstberger R.Nuclear translocation of the transcription factor STAT5in the rat brain after systemic leptin administration[J].Neuroscience Letters,2007,417:286.

20 Buitenhuis M,Coffer PJ,Koenderman L.Signal transducer and activator of transcription 5(STAT5)[J].Int J Biochem Cell Biol,2004,36:2120.

21 Gu L,Dagvadorj A,Lutz J,et al.Transcription factor Stat3stimulates metastatic behavior of human prostate cancer cells in vivo,whereas Stat5bhas a preferential role in the promotion of prostate cancer cell viability and tumor growth[J].The American Journal of Pathology,2010,176:1959.

22 Goenka S.Transcriptional regulation by STAT6[J].Immunologic Research,2011,50:87.

23 Mottok A,Renn C,Willenbrock K,et al.Somatic hypermutation of SOCS1in lymphocyte-predominant Hodgkin lymphoma is accompanied by high JAK2expression and activation of STAT6[J].Blood,2007,110:3387.

24 秦琴,龙洪清,彭冰,等.信号转导子与转录活化子6(STAT 6)在人结肠癌细胞HT-29(SAT6high)、Caco2(STAT 6null)细胞凋亡中的作用[J].南昌大学学报:医学版,2011,25:1.