腺苷酸依赖的蛋白激酶在子宫腺肌症中的表达及其作用
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摘要
目的通过检测腺苷酸依赖的蛋白激酶(AMPK)在子宫腺肌症患者在位子宫内膜组织中的表达和定位,探讨AMPK在子宫腺肌症发病中的作用。方法免疫组织化学染色法检测子宫腺肌症患者在位子宫内膜组织(20例)和对照组正常子宫内膜(20例)中AMPK的表达情况; Real-time PCR检测子宫腺肌症患者在位子宫内膜组织和对照组正常子宫内膜中AMPK、M2型丙酮酸激酶(PKM2) mRNA表达水平; Western blotting检测子宫腺肌症患者在位子宫内膜组织和对照组正常子宫内膜中AMPK、PKM2蛋白表达水平。结果 AMPK主要在子宫腺肌症患者在位子宫内膜组织和对照组正常子宫内膜中的腺上皮细胞质中表达,免疫组化结果显示子宫腺肌症患者在位子宫内膜腺上皮细胞中AMPK表达在分泌期和增殖期均高于正常子宫内膜(P <0. 001),而对照组、实验组相应的增殖期和分泌期中AMPK表达差异无统计学意义(P均> 0. 05)。子宫腺肌症患者在位内膜组织中AMPK、PKM2mRNA表达水平和蛋白表达水平均高于正常子宫内膜,差异有统计学意义(P <0. 001),而对照组、实验组相应的增殖期和分泌期中AMPK、PKM2 mRNA表达水平和蛋白表达水平差异无统计学意义(P>0. 05)。结论 AMPK可能通过促进糖酵解代谢在子宫腺肌症的发生发展中发挥作用。
Objective To explore the role of adenosine monophosphate dependent protein kinase( AMPK) in the pathogenesis of adenomyosis by investigating its expression and localization in the eutopic endometrial tissues in adenomyosis.Methods The localizations of AMPK in eutopic adenomyosis endometrium( 20 cases) and normal endometrium( 20 cases) were detected with immunohistochemistry. The mRNA and protein expressions of AMPK and M2 pyruvate kinase( PKM2) were determined with Real-time PCR and Western blotting,respectively. Results AMPK was mainly expressed in the glandular epithelial cytoplasm in normal endometrium and eutopic adenomyosis endometrium. Immunohistochemistry showed that AMPK expression was higher in eutopic adenomyosis endometrium than in normal endometrium either in the secretory or proliferative phases( P < 0. 001),while there were no statistical differences in the expression of AMPK in the proliferative phase and secretory phase of each group( all P > 0. 05). The mRNA and protein expressions of AMPK and PKM2 were higher in eutopic adenomyosis endometrium than in normal endometrium( P <0.001),while there were no statistical differences in the mRNA and protein expressions of adenosine monophosphate and PKM2in the proliferative phase and secretory phase of each group( all P > 0. 05). Conclusion AMPK promotes glycolysis and plays an important role in the genesis and development of adenomyosis.
Objective To explore the role of adenosine monophosphate dependent protein kinase( AMPK) in the pathogenesis of adenomyosis by investigating its expression and localization in the eutopic endometrial tissues in adenomyosis.Methods The localizations of AMPK in eutopic adenomyosis endometrium( 20 cases) and normal endometrium( 20 cases) were detected with immunohistochemistry. The mRNA and protein expressions of AMPK and M2 pyruvate kinase( PKM2) were determined with Real-time PCR and Western blotting,respectively. Results AMPK was mainly expressed in the glandular epithelial cytoplasm in normal endometrium and eutopic adenomyosis endometrium. Immunohistochemistry showed that AMPK expression was higher in eutopic adenomyosis endometrium than in normal endometrium either in the secretory or proliferative phases( P < 0. 001),while there were no statistical differences in the expression of AMPK in the proliferative phase and secretory phase of each group( all P > 0. 05). The mRNA and protein expressions of AMPK and PKM2 were higher in eutopic adenomyosis endometrium than in normal endometrium( P <0.001),while there were no statistical differences in the mRNA and protein expressions of adenosine monophosphate and PKM2in the proliferative phase and secretory phase of each group( all P > 0. 05). Conclusion AMPK promotes glycolysis and plays an important role in the genesis and development of adenomyosis.
引文
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[15]Possik E,Ajisebutu A,Manteghi S,et al. FLCN andAM PK confer resistance to hyperosmotic stress via re-modeling of glycogen stores[J]. PLoS Genet,2015,11(10):e1005520. doi:10. 1371/journal. pgen. 1005520.
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[18]Faubert B,Boily G,Izreig S,et al. AMPK is a negativeregulator of the Warburg effect and suppresses tumorgrow th in vivo[J]. Cell M etab,2013,17(1):113-124.
[19] Wiese EK,Hitosugi T. Tyrosine Kinase Signaling inCancer M etabolism:PKM2 Paradox in the WarburgEffect[J]. Front Cell Dev Biol,2018,6:79. doi:10.3389/fcell. 2018. 00079.
[20]Zheng X,Zhou Y,Chen W,et al. Ginsenoside 20(S)-Rg3 prevents PKM2-targeting mir-324-5p from H19sponging to antagonize the Warburg Effect in ovariancancer cells[J]. Cell Physiol Biochem,2018,51(3):1340-1353.
[21]Zhou L,Jiang S,Fu Q,et al. FASN,Erb B2-mediatedglycolysis is required for breast cancer cell migration[J]. Oncol Rep,2016,35(5):2715-2722.
[22]Yan XL,Zhang XB,Ao R,et al. Effects of shRNA-mediated silencing of PKM2 gene on aerobic glycolysis,cell migration,cell invasion,and apoptosis in colorectalcancer cells[J]. J Cell Biochem,2017,118(12):4792-4803.
[23]Chakraborty PK,Mustafi SB,Xiong X,et al. MICU1drives glycolysis and chemoresistance in ovarian cancer[J]. Nat Commun,2017,8:14634. doi:10. 1038/ncomms14634.
[24]Jin L,Chun J,Pan C,et al. Phosphorylation-mediatedactivation of LDHA promotes cancer cell invasion andtumour metastasis[J]. Oncogene,2017,36(27):3797-3806.
[25]Li L,Zhu L,Hao B,et al. i NOS-derived nitric oxidepromotes glycolysis by inducing pyruvate kinase M2 nu-clear translocation in ovarian cancer[J]. Oncotarget,2017,8(20):33047-33063.
[2]Pontis A,D'Alterio MN,Pirarba S,et al. Adenomyosis:a systematic review of medical treatment[J]. GynecolEndocrinol,2016,32(9):696-700.
[3]熊小琴,布占红,朱虹,等.部分与全部子宫内膜电切术治疗子宫腺肌症临床疗效观察[J].中国研究型医院,2018,5(5):41-43.XIONG Xiaoqin,BU Zhanhong,ZHU Hong,et al. Clin-ical efficacy of partial and total endometrial resection foradenomyosis[J]. Journal of Chinese Research Hospitals,2018,5(5):41-43.
[4] Vannuccini S,Luisi S,Tosti C,et al. Role of medicaltherapy in the management of uterine adenomyosis[J].Fertil Steril,2018,109(3):398-405.
[5] Vannuccini S,Tosti C,Carmona F,et al. Pathogenesisof adenomyosis:an update on molecular mechanisms[J]. Reprod Biomed Online,2017,35(5):592-601.
[6]Jeon SM. Regulation and function of AMPK in physiolo-gy and diseases[J]. Exp M ol M ed,2016,48(7):e245.doi:10. 1038/emm. 2016. 81.
[7] Villanueva-Paz M,Cotán D,Garrido-Maraver J,et al.AM PK regulation of cell grow th,apoptosis,autophagy,and bioenergetics[J]. Exp Suppl,2016,107:45-71.doi:10. 1007/978-3-319-43589-3_3.
[8]Kim N,Lee JO,Lee HJ,et al. AMPK,a metabolic sen-sor,is involved in isoeugenol-induced glucose uptake inmuscle cells[J]. J Endocrinol,2016,228(2):105-114.
[9]Huang X,Li X,Xie X,et al. High expressions of LDHAand AM PK as prognostic biomarkers for breast cancer[J]. Breast,2016,30:39-46. doi:10. 1016/j. breast.2016. 08. 014.
[10] Younes G,Tulandi T. Effects of adenomyosis on invitro fertilization treatment outcomes:a meta-analysis[J]. Fertil Steril,2017,108(3):483-490.
[11]Dueholm M. Uterine adenomyosis and infertility,reviewof reproductive outcome after in vitro fertilization andsurgery[J]. Acta Obstet Gynecol Scand,2017,96(6):715-726.
[12]Kishi Y,Shimada K,Fujii T,et al. Phenotypic charac-terization of adenomyosis occurring at the inner and out-er myometrium[J]. PLoS One, 2017, 12(12):e0189522. doi:10. 1371/journal. pone. 0189522.
[13]Ha J,Guan KL,Kim J. AMPK and autophagy in glu-cose/glycogen metabolism[J]. M ol Aspects M ed,2015,46:46-62. doi:10. 1016/j. mam. 2015. 08. 002.
[14]Cokorinos EC,Delmore J,Reyes AR,et al. Activationof skeletal muscle AM PK promotes glucose disposal andglucose low ering in non-human primates and mice[J].Cell Metab,2017,25(5):1147-1159. e10. doi:10.1016/j. cmet. 2017. 04. 010.
[15]Possik E,Ajisebutu A,Manteghi S,et al. FLCN andAM PK confer resistance to hyperosmotic stress via re-modeling of glycogen stores[J]. PLoS Genet,2015,11(10):e1005520. doi:10. 1371/journal. pgen. 1005520.
[16]San-Millán I,Brooks GA. Reexamining cancer metabo-lism:lactate production for carcinogenesis could be thepurpose and explanation of the Warburg Effect[J].Carcinogenesis,2017,38(2):119-133.
[17]Tekade RK,Sun X. The Warburg effect and glucose-de-rived cancer theranostics[J]. Drug Discov Today,2017,22(11):1637-1653.
[18]Faubert B,Boily G,Izreig S,et al. AMPK is a negativeregulator of the Warburg effect and suppresses tumorgrow th in vivo[J]. Cell M etab,2013,17(1):113-124.
[19] Wiese EK,Hitosugi T. Tyrosine Kinase Signaling inCancer M etabolism:PKM2 Paradox in the WarburgEffect[J]. Front Cell Dev Biol,2018,6:79. doi:10.3389/fcell. 2018. 00079.
[20]Zheng X,Zhou Y,Chen W,et al. Ginsenoside 20(S)-Rg3 prevents PKM2-targeting mir-324-5p from H19sponging to antagonize the Warburg Effect in ovariancancer cells[J]. Cell Physiol Biochem,2018,51(3):1340-1353.
[21]Zhou L,Jiang S,Fu Q,et al. FASN,Erb B2-mediatedglycolysis is required for breast cancer cell migration[J]. Oncol Rep,2016,35(5):2715-2722.
[22]Yan XL,Zhang XB,Ao R,et al. Effects of shRNA-mediated silencing of PKM2 gene on aerobic glycolysis,cell migration,cell invasion,and apoptosis in colorectalcancer cells[J]. J Cell Biochem,2017,118(12):4792-4803.
[23]Chakraborty PK,Mustafi SB,Xiong X,et al. MICU1drives glycolysis and chemoresistance in ovarian cancer[J]. Nat Commun,2017,8:14634. doi:10. 1038/ncomms14634.
[24]Jin L,Chun J,Pan C,et al. Phosphorylation-mediatedactivation of LDHA promotes cancer cell invasion andtumour metastasis[J]. Oncogene,2017,36(27):3797-3806.
[25]Li L,Zhu L,Hao B,et al. i NOS-derived nitric oxidepromotes glycolysis by inducing pyruvate kinase M2 nu-clear translocation in ovarian cancer[J]. Oncotarget,2017,8(20):33047-33063.