• 1.大連醫(yī)科大學(xué)附屬第一醫(yī)院普外二科(遼寧大連 116011);;
  • 2.大連醫(yī)科大學(xué)附屬第一醫(yī)院中心實(shí)驗(yàn)室(遼寧大連 116011);

目的 探討K-ras突變多肽致敏樹突狀細(xì)胞(DC)對(duì)細(xì)胞趨化因子CCL19、CCL22和細(xì)胞骨架蛋白fascin-1表達(dá)的影響。方法 聯(lián)合應(yīng)用重組人粒-巨噬細(xì)胞集落刺激因子和白細(xì)胞介素(IL)-4誘導(dǎo)培養(yǎng)外周血DC，收集培養(yǎng)7 d后的DC并分為未負(fù)載組（加入RPMI 1640培養(yǎng)液50 μg/ml)和K-ras負(fù)載組（加入K-ras突變多肽50 μg/ml）。流式細(xì)胞儀測(cè)定負(fù)載前、后DC表面標(biāo)志CD1a、CD80及CD86； 掃描電鏡和透射電鏡觀察負(fù)載K-ras突變多肽后的DC形態(tài)結(jié)構(gòu)； ELISA法檢測(cè)2組DC培養(yǎng)上清液中IL-12、CCL19和CCL22的表達(dá)； Western blot法測(cè)定2組DC骨架蛋白fascin-1的表達(dá)。結(jié)果 ①K-ras突變多肽負(fù)載DC后，CD1a、CD80及CD86表面分子表達(dá)率明顯高于負(fù)載前(P lt;0.01)。②掃描電鏡下見負(fù)載后的DC呈花瓣?duì)睢渲油黄穑?透射電鏡下見負(fù)載后的DC形態(tài)非常不規(guī)則，樹枝狀或毛刺狀的突起明顯增多。③K-ras負(fù)載組負(fù)載后不同時(shí)相(6、12、24及48 h）的IL-12、CCL19及CCL22的表達(dá)水平明顯高于未負(fù)載組(P lt;0.01)。④K-ras負(fù)載組fascin-1蛋白的表達(dá)水平也高于未負(fù)載組(P lt;0.01)。
結(jié)論 K-ras突變多肽能夠促進(jìn)DC成熟，并使細(xì)胞趨化因子和骨架蛋白表達(dá)水平增加，能夠增強(qiáng)DC游走遷移。

引用本文： 江良縣,譚廣,王忠裕,竇春鵬,單路娟. K-ras突變多肽致敏樹突狀細(xì)胞對(duì)CCL19、CCL22和fascin-1表達(dá)的影響. 中國(guó)普外基礎(chǔ)與臨床雜志, 2010, 17(9): 917-921. doi: 復(fù)制

1. Santegoets SJ, van den Eertwegh AJ, van de Loosdrecht AA, et al. Human dendritic cell line models for DC differentiation and clinical DC vaccination studies [J]. J Leukoc Biol, 2008; 84(6): 13641373.
2. Kim HS, Choo YS, Koo T, et al. Enhancement of antitumor immunity of dendritic cells pulsed with heattreated tumor lysate in murine pancreatic cancer [J]. Immunol Lett, 2006; 103(2): 142148.
3. Xia Y, Dai J, Lu PR, et al. Distinct effect of CD40 and TNFsignaling on the chemokine/chemokine receptor expression and function of the human monocytederived dendritic cells [J]. Cell Mole Immunol, 2008; 5(2)： 4656.
4. Ishimoto H, Yanagihara K, Araki N, et al. Singlecell observation of phagocytosis by human blood dendritic cells [J]. Jpn J Infect Dis, 2008; 61(4): 294297.
5. Koido S, Hara E, Homma S, et al. Cancer vaccine by fusions of dendritic and cancer cells [J].Clin Dev Immunol, 2009; 2009: ID657369. doi: 10.1155/2009/657369.
6. Huang CF, Monie A, Weng WH, et al. DNA vaccines for cervical cancer [J]. Am J Transl Res, 2010; 2(1): 7587.
7. 薛剛， 曹永寬， 王培紅， 等. 腫瘤細(xì)胞裂解物致敏的樹突狀細(xì)胞對(duì)結(jié)腸癌LoVo細(xì)胞的殺傷作用 [J]. 中國(guó)普外基礎(chǔ)與臨床雜志， 2009； 16(1)： 4851.
8. 譚廣， 任金帥， 王忠裕， 等. TNFα增強(qiáng)樹突狀細(xì)胞誘導(dǎo)的抗胰腺癌免疫應(yīng)答的實(shí)驗(yàn)研究 [J]. 中國(guó)普外基礎(chǔ)與臨床雜志， 2006； 13(6)： 689692.
9. Lee DH. Dendritic cellsbased vaccine and immune monitoring for hepatocellular carcinoma [J]. Korean J Physiol Pharmacol, 2010; 14(1): 1114.
10. Srivastava PK. Therapeutic cancer vaccines [J]. Curr Opin Immunol, 2006; 18(2): 201205.
11. Kikuchi T. Genetically modified dendritic cells for therapeutic immunity [J]. Tohoku J Exp Med, 2006; 208(1): 18.
12. Recktenwald CV, Kellner R, Lichtenfels R, et al. Altered detoxification status and increased resistance to oxidative stress by Kras transformation [J]. Cancer Res, 2008; 68(24): 1008610093.
13. Shen YM, Yang XC, Yang C, et al. Enhanced therapeutic effects for human pancreatic cancer by application Kras and IGFIR antisense oligodeoxynucleotides [J]. World J Gastroenterol, 2008; 14(33): 51765185.
14. Lee SH, Lee SJ, Jung YS, et al. Blocking of p53Snail binding, promoted by oncogenic KRas, recovers p53 expression and function [J]. Neoplasia, 2009; 11(1): 2231.
15. 楊波， 何楊， 孫冬林， 等. Kras(12Val)樹突狀細(xì)胞疫苗誘導(dǎo)抗胰腺癌免疫反應(yīng) [J]. 蘇州大學(xué)學(xué)報(bào)（醫(yī)學(xué)版）, 2007； 27（5）： 691700.
16. Wang YX, Gao L, Ji ZZ. Inhibitary effects of antisense oligonucleotide specific to Kras point mutation on the target gene expression in human pancreatic carcinoma cells [J]. Chin Med J (Engl), 2007; 120(16): 14481450.
17. Biragyn A, Ruffini PA, Coscia M, et al. Chemokine receptormediated delivery directs selftumor antigen efficiently into the class Ⅱ processing pathway in vitro and induces protective immunity in vivo [J]. Blood, 2004; 104(7): 19611969.
18. Bertho N, Adamski H, Toujas L, et al. Efficient migration of dendritic cells toward lymph node chemokines and induction of T(H)1 responses require maturation stimulus and apoptotic cell interaction [J]. Blood, 2005; 106(5): 17341741.
19. Trinchieri G. Interleukin12 and the regulation of innate resistance and adaptive immunity [J]. Nat Rev Immunol, 2003; 3(2): 133146.
20. Brivio F, Lissoni P, Rovelli F, et al. Effects of IL2 preoperative immunotherapy on surgeryinduced changes in angiogenic regulation and its prevention of VEGF increase and IL12 decline [J]. Hepatogastroenterology, 2002; 49(44): 385387.
21. Pietil TE, Veckman V, Lehtonen A, et al. Multiple NFkappaB and IFN regulatory factor family transcription factors regulate CCL19 gene expression in human monocytederived dendritic cells [J]. J Immunol, 2007; 178(1): 253261.
22. Nakano H, Lin KL, Yanagita M, et al. Bloodderived inflammatory dendritic cells in lymph nodes stimulate acute TH1 immune responses [J]. Nat Immunol, 2009; 10(4): 394402.
23. Britschgi MR, Link A, Lissandrin TK, et al. Dynamic modulation of CCR7 expression and function on naive T lymphocytes in vivo [J]. J Immunol, 2008; 181(11): 76817688.
24. 李立, 張升寧, 冉江華, 等. 受體來源未成熟樹突狀細(xì)胞誘導(dǎo)大鼠肝移植免疫低反應(yīng)性的實(shí)驗(yàn)研究 [J]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2009; 16(1): 3238.
25. Yamashita U, Kuroda E. Regulation of macrophagederived chemokine (MDC, CCL22) production [J]. Crit Rev Immunol, 2002; 22(2): 105114.
26. Gobert M, Treilleux I, BendrissVermare N, et al. Regulatory T cells recruited through CCL22/CCR4 are selectively activated in lymphoid infiltrates surrounding primary breast tumors and lead to an adverse clinical outcome [J]. Cancer Res, 2009; 69(5): 20002009.
27. Mailloux AW, Young MR. NKdependent increases in CCL22 secretion selectively recruits regulatory T cells to the tumor microenvironment [J]. J Immunol, 2009; 182(5): 27532765.
28. AlAlwan MM, Rowden G, Lee TD, et al. Fascin is involved in the antigen presentation activity of mature dendritic cells [J]. J Immunol, 2001; 166(1): 338345.
29. Geyeregger R, Zeyda M, Bauer W, et al. Liver X receptors regulate dendritic cell phenotype and function through blocked induction of the actinbundling protein fascin [J]. Blood, 2007; 109(10): 42884295.
30. Zanet J, Payre F, Plaza S. Fascin for cell migration in Drosophila [J]. Fly (Austin), 2009; 3(4): 281282.
31. Hashimoto Y, Loftis DW, Adams JC. Fascin1 promoter activity is regulated by CREB and the aryl hydrocarbon receptor in human carcinoma cells [J]. PLoS One, 2009; 4(4): e5130.
  1. 1. Santegoets SJ, van den Eertwegh AJ, van de Loosdrecht AA, et al. Human dendritic cell line models for DC differentiation and clinical DC vaccination studies [J]. J Leukoc Biol, 2008; 84(6): 13641373.
  2. 2. Kim HS, Choo YS, Koo T, et al. Enhancement of antitumor immunity of dendritic cells pulsed with heattreated tumor lysate in murine pancreatic cancer [J]. Immunol Lett, 2006; 103(2): 142148.
  3. 3. Xia Y, Dai J, Lu PR, et al. Distinct effect of CD40 and TNFsignaling on the chemokine/chemokine receptor expression and function of the human monocytederived dendritic cells [J]. Cell Mole Immunol, 2008; 5(2)： 4656.
  4. 4. Ishimoto H, Yanagihara K, Araki N, et al. Singlecell observation of phagocytosis by human blood dendritic cells [J]. Jpn J Infect Dis, 2008; 61(4): 294297.
  5. 5. Koido S, Hara E, Homma S, et al. Cancer vaccine by fusions of dendritic and cancer cells [J].Clin Dev Immunol, 2009; 2009: ID657369. doi: 10.1155/2009/657369.
  6. 6. Huang CF, Monie A, Weng WH, et al. DNA vaccines for cervical cancer [J]. Am J Transl Res, 2010; 2(1): 7587.
  7. 7. 薛剛， 曹永寬， 王培紅， 等. 腫瘤細(xì)胞裂解物致敏的樹突狀細(xì)胞對(duì)結(jié)腸癌LoVo細(xì)胞的殺傷作用 [J]. 中國(guó)普外基礎(chǔ)與臨床雜志， 2009； 16(1)： 4851.
  8. 8. 譚廣， 任金帥， 王忠裕， 等. TNFα增強(qiáng)樹突狀細(xì)胞誘導(dǎo)的抗胰腺癌免疫應(yīng)答的實(shí)驗(yàn)研究 [J]. 中國(guó)普外基礎(chǔ)與臨床雜志， 2006； 13(6)： 689692.
  9. 9. Lee DH. Dendritic cellsbased vaccine and immune monitoring for hepatocellular carcinoma [J]. Korean J Physiol Pharmacol, 2010; 14(1): 1114.
  10. 10. Srivastava PK. Therapeutic cancer vaccines [J]. Curr Opin Immunol, 2006; 18(2): 201205.
  11. 11. Kikuchi T. Genetically modified dendritic cells for therapeutic immunity [J]. Tohoku J Exp Med, 2006; 208(1): 18.
  12. 12. Recktenwald CV, Kellner R, Lichtenfels R, et al. Altered detoxification status and increased resistance to oxidative stress by Kras transformation [J]. Cancer Res, 2008; 68(24): 1008610093.
  13. 13. Shen YM, Yang XC, Yang C, et al. Enhanced therapeutic effects for human pancreatic cancer by application Kras and IGFIR antisense oligodeoxynucleotides [J]. World J Gastroenterol, 2008; 14(33): 51765185.
  14. 14. Lee SH, Lee SJ, Jung YS, et al. Blocking of p53Snail binding, promoted by oncogenic KRas, recovers p53 expression and function [J]. Neoplasia, 2009; 11(1): 2231.
  15. 15. 楊波， 何楊， 孫冬林， 等. Kras(12Val)樹突狀細(xì)胞疫苗誘導(dǎo)抗胰腺癌免疫反應(yīng) [J]. 蘇州大學(xué)學(xué)報(bào)（醫(yī)學(xué)版）, 2007； 27（5）： 691700.
  16. 16. Wang YX, Gao L, Ji ZZ. Inhibitary effects of antisense oligonucleotide specific to Kras point mutation on the target gene expression in human pancreatic carcinoma cells [J]. Chin Med J (Engl), 2007; 120(16): 14481450.
  17. 17. Biragyn A, Ruffini PA, Coscia M, et al. Chemokine receptormediated delivery directs selftumor antigen efficiently into the class Ⅱ processing pathway in vitro and induces protective immunity in vivo [J]. Blood, 2004; 104(7): 19611969.
  18. 18. Bertho N, Adamski H, Toujas L, et al. Efficient migration of dendritic cells toward lymph node chemokines and induction of T(H)1 responses require maturation stimulus and apoptotic cell interaction [J]. Blood, 2005; 106(5): 17341741.
  19. 19. Trinchieri G. Interleukin12 and the regulation of innate resistance and adaptive immunity [J]. Nat Rev Immunol, 2003; 3(2): 133146.
  20. 20. Brivio F, Lissoni P, Rovelli F, et al. Effects of IL2 preoperative immunotherapy on surgeryinduced changes in angiogenic regulation and its prevention of VEGF increase and IL12 decline [J]. Hepatogastroenterology, 2002; 49(44): 385387.
  21. 21. Pietil TE, Veckman V, Lehtonen A, et al. Multiple NFkappaB and IFN regulatory factor family transcription factors regulate CCL19 gene expression in human monocytederived dendritic cells [J]. J Immunol, 2007; 178(1): 253261.
  22. 22. Nakano H, Lin KL, Yanagita M, et al. Bloodderived inflammatory dendritic cells in lymph nodes stimulate acute TH1 immune responses [J]. Nat Immunol, 2009; 10(4): 394402.
  23. 23. Britschgi MR, Link A, Lissandrin TK, et al. Dynamic modulation of CCR7 expression and function on naive T lymphocytes in vivo [J]. J Immunol, 2008; 181(11): 76817688.
  24. 24. 李立, 張升寧, 冉江華, 等. 受體來源未成熟樹突狀細(xì)胞誘導(dǎo)大鼠肝移植免疫低反應(yīng)性的實(shí)驗(yàn)研究 [J]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2009; 16(1): 3238.
  25. 25. Yamashita U, Kuroda E. Regulation of macrophagederived chemokine (MDC, CCL22) production [J]. Crit Rev Immunol, 2002; 22(2): 105114.
  26. 26. Gobert M, Treilleux I, BendrissVermare N, et al. Regulatory T cells recruited through CCL22/CCR4 are selectively activated in lymphoid infiltrates surrounding primary breast tumors and lead to an adverse clinical outcome [J]. Cancer Res, 2009; 69(5): 20002009.
  27. 27. Mailloux AW, Young MR. NKdependent increases in CCL22 secretion selectively recruits regulatory T cells to the tumor microenvironment [J]. J Immunol, 2009; 182(5): 27532765.
  28. 28. AlAlwan MM, Rowden G, Lee TD, et al. Fascin is involved in the antigen presentation activity of mature dendritic cells [J]. J Immunol, 2001; 166(1): 338345.
  29. 29. Geyeregger R, Zeyda M, Bauer W, et al. Liver X receptors regulate dendritic cell phenotype and function through blocked induction of the actinbundling protein fascin [J]. Blood, 2007; 109(10): 42884295.
  30. 30. Zanet J, Payre F, Plaza S. Fascin for cell migration in Drosophila [J]. Fly (Austin), 2009; 3(4): 281282.
  31. 31. Hashimoto Y, Loftis DW, Adams JC. Fascin1 promoter activity is regulated by CREB and the aryl hydrocarbon receptor in human carcinoma cells [J]. PLoS One, 2009; 4(4): e5130.