神经示踪研究背景cankao

1、全反式维甲酸对正常宫颈上皮及宫颈癌 HeLa 细胞的影响探究 ATRA 作用下宫颈癌HeLa 细胞的 Cx43、Cx26 的表达及细胞侵袭、转移能力及 ATRA 对正常宫颈上皮的影响宫颈癌宫颈癌是在全球女性中居于前位的常见肿瘤，恶性程度高，许多患者在发现时已处于晚期。1papilloma，HPV）2 目前公认的研究结果表明，某些型的人瘤（human是宫颈癌的主要致病。HPV 与宫颈上皮的储备细胞（cervicalcells，CERCs：基底细胞，具有分化潜能）上的 HPV 受体结合，通过泛素thelial化降解 P53 和 Rb 蛋白2,3 HDACi 4-5，激活端粒酶活性4 HDACi 6

2、，下调细胞 RAR2 和E-cadherin 等黏附分子的表达，促进上皮间质转化（thelial-mesenchymal transition，EMT）。5,6,7 HDACi 7,8,9 继而引起 CERCs 的分化成熟受阻，出现分化与凋亡的双重并出现永生化生长趋势，CERCs 进入异常细胞周期，发生异常增生和化生、瘤变、致恶性转化。8,9 HDACi10,11 临床病理从宫颈上皮异常增生和化生发展到位癌、浸润癌甚至播散转移。颈上皮内瘤变（CIN）然后发展为原1, 等. 山东省 1970-2005 年宫颈癌率变化趋势 J.中国卫生统计, 2009, 26 (4): 683.Shaikh F,

3、 Sanchi P, Rawal R: Molecular screening of compounds to the predicted Protein-Proteineraction site of Rb1-E7 with p53-E6 in HPV J. Bioinformation, 2012, 8:670-612.Maher DM, Bell MC, ODonnell EA, Gupta BK, Jaggi M, Chauhan SC: Curcuminsuppresses human papillomaoncoproteins, restores p53, Rb, and PTPN

4、13proteins and inhibits benzoapyrene-induced upregulation of HPV E7 J. Molecular carcinogenesis, 2011,50:47-57.4 Mighty KK, Laimins LA: The role of human papillomaRecent results in cancer research Fortschritte der Krebsforsces in oncegenesis J.g Progres dans lesrechercher le cancer, 2014, 193:135-14

5、8.5 De-Castro Arce J, Gockel-Krzikalla E, Rosl F: Retinoic acid receptor beta silenhuman papilloma-18 oncogene expresby induction of de novo methylationand heterochromatinization of the viral control region J. The .lournal of biological chemistry, 2007, 282:28520-28529.6 Sun LL, Cao DY,JX, Li H, Zho

6、u XR, Song ZQ, Cheng XM, Chen J, Shen K:Population-based case-control study on DAPKl, RAR-beta2 and MGMT methylation in liquid-based cytology J. Archives of gynecology and obstetrics, 2012,285:1433-1439.7 Zannoni GF, Prisco MCP Vellone VG, De Stefano I, Scambia G, Gallo D:Changeshe expresof oestroge

7、n receptors and E-cadherin as molecularmarkers of progres patientswith 58:265-275.from normalthelium to invasive cancerinelderlyrsquamouscellcarcinoma J. Histopathology, 2011,8 Sun Y, Tokar EJ, Waalkes MP: Overabundance of puive cancer stem cells in human skin keratinocyte cells malignantly transfor

8、med by arsenic J. Toxicologicalscien: an offil journal of the Society of Toxicology, 2012, 125:20-29.9 Mighty KK, Laimins LA: p63 is nesary for the activation of humanpapillomalate viral functions uponthelial differentiation J. Journal ofvirology, 2011, 85:8863-8869.诱导分化治疗与全反式维甲酸（all-trans retinoic

9、acid, ATRA）全反式维甲酸（ATRA）是维生素 A 在内代谢生成的高活性衍生物，对胚胎发育过细胞分化机体生长发育有重要作用。10 HeLa/MMC 1 ATRA 等 RA 类物质在上皮细胞生长发育与分化中也扮演着重要角色，其缺乏可导致上皮分化异常。RA 类物质的作用受维甲酸受体 RARs/RXRs 的调节, 其中RAR2 的编码被认为是重要的抑癌。ATRA 诱导分化的治疗方法的研究较为深入，特别在白血病的治疗上表现突出。11,12,13 白血病细胞分化治疗，HeLa/MMC 2,3 对实体瘤的分化研究也有较多如，结肠癌、胃癌、宫颈癌。已有研究证明，ATRA 可有效抑制宫颈细胞 HeLa

10、 增殖并可诱导其分化，并可提高宫颈癌耐药细胞株 HeLa/MMC 对化疗的敏感性。14 HeLa/MMC 6 对 HeLa/MMC 的侵袭、转移也有抑制作用15 HeLa/MMC 诱导分化治疗可通过药物诱导肿瘤细胞的幼稚状态而向正常分化方向发展，是较为理想的控制肿瘤的治疗方法。但其缺点也较为明显：其毒性作用16藻蓝 8，及导致维甲酸综合症与耐药反应使其在临床的应用受到极大的限制。在正常与异常并存的上应用具性的药物势必对其二者有不同的影响。10 Stavrids MP, Collins BJ, Storey KG. Retinoic acid orchestrates fibroblast gr

11、owth factor signaling to drive embrynonic stem cell differentiation J. Development, 2010, 137(6):881-890.11 主鸿鹄,漆, 等. 维甲酸和砷剂联合诱导急性早幼粒细胞白血病中 PML-RAR 表达上调不影响预 后 J. 中国实验血液学杂志 , 2013,21(4):872-878.12, 等. 全反式维甲酸诱导 NB4 细胞分化过泛素-蛋白酶体系统的表达变化J. 福建医学学报, 2012,46(5):297-300.13物治疗. 白血病细胞的分化治疗C.抗肿瘤药物研究新进展与肿瘤个性化药集,

12、 201314 Shiola G, Ki K. Retinoids and theirgeges in liver functions anddiseases J. J Gastroenterol Hepatol, 2013, 28(suppl 1):33-37.15 胡海燕,. 全反式维甲酸对宫颈癌耐药细胞株 HeLa/MMC 侵袭、转移能力的影响及其分子机制研究J. 中国, 2015, 12(6):18-21.16 Siddikuzzaman, Grace VM. Inhibition of metasicg cancer in C57BL/6micebyliomeencapsulated

13、alltransretinoicacid(ATRA)J.Immunopharmacol, 2012,14(4):570-579.间隙连接蛋白（Connexin，Cx）间隙连接（GJ） 是细胞间的直接通讯方式。两细胞间有连接子（Connexon）存在，连接子由两组 6 聚体间隙连接蛋白（Connexin，Cx）组成。17 四 1 Cx 是细胞间隙连接的基本组成。间隙连接允许信息分子（1kD）交换，能够加速相连或的细胞快速、可逆地对外界信号做出协同反应。间隙连接介导细胞间隙连接通讯（GJIC），其在胚胎发育、细胞分化和增殖、稳定内环境、以及协调多细胞间活动方面具有关键作用。在多种外界条件刺激或病理

14、条件下，Cx 的表达及分部均有改变，一度被认为是抑癌基因。很多剂可以降低 Cx 的表达或抑制 GJ 生成。18,19,20,21 复 12,13,14,15Cx 的过表达可降低细胞对物质的敏感性，抑癌物质则可提高 Cx 表达及 GJIC 功能。22,23 复 16,17在肿瘤组织或细胞中也有发现 GJIC 的缺失或者 Cx 的表达下调。24,25,26,27 复 18,19,20,21因此，肿瘤细胞内过表达的连接子蛋白可引起癌细胞生长缓慢、转移及侵袭能力降低、细胞高分化趋势等变化。25,28,29 复 19,24,25 然而现有研究表明，在某些肿瘤细胞中，Cx 的表达与正常细胞水平接近，甚至能

15、促进肿瘤细胞的侵袭、内渗、外渗和转移等过程。30,31,32复 26,27,28较早的研究结果认为Cx 通过由正常细胞经GJ 进入肿瘤细胞的特殊小分子实现肿瘤抑制作用，被称为 GJIC 依赖机理（GJIC-dependent）。但是越来越多的研究者发现许多在肿瘤细胞内过表达的 Cx 并未定位在细胞膜上形成间隙连接，却能起到抑制肿瘤的作用，提示 GJIC非依赖机理的存在。28,33,34 复 24,30,31 其在于 Cx 通过调控下游分子起到抑制肿瘤发生的作用。已有研究结果者为 Cx43，Cx32。35,36,37,38,39,40 复 32,34,35,36,37,38 因此有理由认为，Cx

16、 在肿瘤发生、发展与转移中的作用因 Cx 种类、组织种类不同而不同，而且与其联系紧密。正常人宫颈组织中存在 Cx26、Cx43 的表达，而宫颈癌细胞系 HeLa 中存在多种 Cx 的表达缺失，且 Cx 表达水平与宫颈癌的恶性程度存在负相关关系。KING419等在研究宫颈癌前病变区域 Cx43 表达时首次发现不典型增生细胞是去间隙连接与宫颈上皮细胞恶性行为有关。华4210等研究进一步表明 Cx43 表达与宫颈癌密切相关。Hagemann43 李16等研究发现 Cx43 变化与宫颈不典型增生程度改变相一致，且其表达先于宫颈的病理改变等研究发现 Cx26 与 Cx43 在不同分化程度的浸润44癌中表

17、达蛋白的程度并不一致，但都与宫颈上皮的异常分化增生有密切关系，提示二者在对宫颈癌细胞生长作用上的机制并不相同。参考文献1, 等. 山东省 1970-2005 年宫颈癌率变化趋势 J.中国卫生统计, 2009, 26 (4): 683.Shaikh F, Sanchi P, Rawal R: Molecular screening of compounds to the predicted Protein-Proteineraction site of Rb1-E7 with p53-E6 in HPV J. Bioinformation, 2012, 8:670-612.Maher DM, B

18、ell MC, ODonnell EA, Gupta BK, Jaggi M, Chauhan SC: Curcuminsuppresses human papillomaoncoproteins, restores p53, Rb, and PTPN13proteins and inhibits benzoapyrene-induced upregulation of HPV E7 J. Molecular carcinogenesis, 2011,50:47-57.4 Mighty KK, Laimins LA: The role of human papillomaRecent resu

19、lts in cancer research Fortschritte der Krebsforsces in oncegenesis J.g Progres dans lesrechercher le cancer, 2014, 193:135-148.5 De-Castro Arce J, Gockel-Krzikalla E, Rosl F: Retinoic acid receptor beta silenhuman papilloma-18 oncogene expresby induction of de novo methylationand heterochromatiniza

20、tion of the viral control region J. The .lournal of biological chemistry, 2007, 282:28520-28529.6 Sun LL, Cao DY,JX, Li H, Zhou XR, Song ZQ, Cheng XM, Chen J, Shen K:Population-based case-control study on DAPKl, RAR-beta2 and MGMT methylation in liquid-based cytology J. Archives of gynecology and ob

21、stetrics, 2012,285:1433-1439.7 Zannoni GF, Prisco MCP Vellone VG, De Stefano I, Scambia G, Gallo D:Changeshe expresof oestrogen receptors and E-cadherin as molecularmarkers of progres patientswith 58:265-275.from normalthelium to invasive cancerinelderlyrsquamouscellcarcinoma J. Histopathology, 2011

22、,8 Sun Y, Tokar EJ, Waalkes MP: Overabundance of puive cancer stem cells in human skin keratinocyte cells malignantly transformed by arsenic J. Toxicologicalscien: an offil journal of the Society of Toxicology, 2012, 125:20-29.9 Mighty KK, Laimins LA: p63 is nesary for the activation of humanpapillo

23、malate viral functions uponthelial differentiation J. Journal ofvirology, 2011, 85:8863-8869.10 Stavrids MP, Collins BJ, Storey KG. Retinoic acid orchestrates fibroblast growth factor signaling to drive embrynonic stem cell differentiation J. Development, 2010, 137(6):881-890.11 主鸿鹄,漆, 等. 维甲酸和砷剂联合诱导

24、急性早幼粒细胞白血病中PML-RAR 表达上调不影响预后J.中国实验血液学杂志, 2013,21(4):872-878.12, 等. 全反式维甲酸诱导 NB4 细胞分化过泛素-蛋白酶体系统的表达变化J. 福建医学学报, 2012,46(5):297-300.13物治疗. 白血病细胞的分化治疗C.抗肿瘤药物研究新进展与肿瘤个性化药集, 201314 Shiola G, Ki K. Retinoids and theirgeges in liver functions anddiseases J. J Gastroenterol Hepatol, 2013, 28(suppl 1):33-37.1

25、5 胡海燕,. 全反式维甲酸对宫颈癌耐药细胞株 HeLa/MMC 侵袭、转移能力的影响及其分子机制研究J. 中国, 2015, 12(6):18-21.16 Siddikuzzaman, Grace VM. Inhibition of metasicg cancer in C57BL/6mice by liome encapsulated all trans retinoic acid (ATRA)J.Immunopharmacol, 2012,14(4):570-579.17 White TW, Bruzzone R, Paul DL. The connexin family ofercel

26、lular channelforming proteins.Kidney. 1995:48, 1148-1157.18 Budunova I, Williams G. Cell culture assays for chemicals with tumor-promotingor tumor-inhibiting activity based on the modulation of J. Cell biology and toxicology, 1994, 10(2): 71-116.19 Fiorini C, Gilleron J, Carette D. et al. Accelerate

27、dercellular communicationernalization of functionalmembrane proteins (connexin 43, N-cadherin and ZO-1) within endocytic vacuoles:an early event oT carcinogenicity J. Biochimica et Biophysica Acta(BBA)-Biomembranes, 2008, 1778(1): 56-67.20 Banerjee D, Das S, Molina SA. et al. Investigation of the re

28、ciprocal relationshipbetn the expresof two gap junction connexin proteins, connexin46 andconnexin43 J. Journal of Biological Chemistry, 2011, 286(27): 24519.21 Choi SJ, Choung YH. Preventive Effect of Green Tea Extracts on the Inhibitionof ConnexinExpresInduced by Carcinogen H202 J. Korean Journal o

29、fOtorhinolaryngology-Head and Neck Surgery, 2008, 51(4): 355-362.22 Wang X, Ramirez A, Budunova I. Overexpresof connexin26he basalkeratinocytes redusensitivity to tumor promoter TPA J. Experimentaldermatology, 2010, 19(7):633-640. 23King TJ, Bertram JS. Connexins ass for cancer chemoprevention andch

30、emotherapy J. Biochimica et Biophysica Acta (BBA)-Biomembranes, 2005, 1719(1-2): 146-160.24 Mesnil M, Crespin S, Avanzo JL. et al. Defective gap functionalercellularcommunicationhe carcinogenic pros J. Biochimica et BiophysicaActa(BBA)-Biomembranes, 2005,1719(1-2): 125-145.Naus CC, Laird DW. Implica

31、tions and challenges of connexin connections to cancer J.Nature Reviews Cancer, 2010, 10(6): 435-441.Yamasaki H, Naus CCG. COMMENTARY: Role of connexin genes in growth control J. Carcinogenesis, 1996, 17(6): 1199.Cronier L, Crespin S, Strale PO. et al. Gap junctions and cancer: new functions for an

32、oldstory J. Antioxidants & redox signaling, 2009, 11(2): 323-338.28 McLachlan E, Shao Q, Wang H. et al. Connexinas tumor suppressors inthree-dimenal mammary cellanoids by regulating differentiation and angiogenesis J.Cancer research, 2006, 66(20): 9886.29 Li Z, Zhou Z, Welch DR. et al. Expressing co

33、nnexin 43 in breast cancer cells redutheirmetastasis togs J. Clinical and Experimental Metastasis, 2008, 25(8): 893-901.30 Dobrowolski R, Sasse P, Schrickel JW. et al. The conditional connexin43G138R mousemutant represents a new mof hereditary oculodentodigital dysplasia in humans J. Humanmolecular

34、genetics, 2008, 17(4): 539.31 Kawasaki Y, Omori Y, Li Q. et al. Cytoplasmic accumulation of connexin32 expands cancerstem cell population in human HuH7 hepatoma cells by enhancing iternational Journal of Cancer, 2011, 128(1): 51-62.frenewal J.32 Elzarrad MK, Haroon A, Willecke K. et al. Connexin-43

35、upregulationicrometastases andtumor vasculature and its rolemedicine, 2008, 6(1): 20.umor cell attaent to pulmonary endothelium J. BMCJiang JX, Gu S. Gap junction-and hemichannel-independent actions of connexins J.Biochimica et Biophysica Acta (BBA)-Biomembranes, 2005, 171 I (2): 208-214.Zhang Y W,

36、Kaneda M, Morita, I. The gap junction-independent tumor-suppressing effect of connexin 43 J. Journal of Biological Chemistry, 2003, 278(45): 44852.35 Gellhaus A, Dong X, Propson S. et al. Connexin43Biological Chemistry, 2004, 279(35): 36931.eracts with NOV J. Journal of36 Leithe, E, Kjense, Sirnes S. et al. Ubiquitylation of the gap junction protein connexin-43signals its trafficking from early endosomes to lysosomes in a proTsg101 J. Journal of cell science, 2009, 122(21): 3883.s mediated by Hrs an