• 1.大連醫(yī)科大學附屬第一醫(yī)院普外二科(遼寧大連 116011);;
  • 2.大連醫(yī)科大學附屬第一醫(yī)院中心實驗室(遼寧大連 116011);

目的 探討K-ras突變多肽致敏樹突狀細胞(DC)對細胞趨化因子CCL19、CCL22和細胞骨架蛋白fascin-1表達的影響。方法 聯合應用重組人粒-巨噬細胞集落刺激因子和白細胞介素(IL)-4誘導培養(yǎng)外周血DC，收集培養(yǎng)7 d后的DC并分為未負載組（加入RPMI 1640培養(yǎng)液50 μg/ml)和K-ras負載組（加入K-ras突變多肽50 μg/ml）。流式細胞儀測定負載前、后DC表面標志CD1a、CD80及CD86； 掃描電鏡和透射電鏡觀察負載K-ras突變多肽后的DC形態(tài)結構； ELISA法檢測2組DC培養(yǎng)上清液中IL-12、CCL19和CCL22的表達； Western blot法測定2組DC骨架蛋白fascin-1的表達。結果 ①K-ras突變多肽負載DC后，CD1a、CD80及CD86表面分子表達率明顯高于負載前(P lt;0.01)。②掃描電鏡下見負載后的DC呈花瓣狀、樹枝樣突起； 透射電鏡下見負載后的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負載組負載后不同時相(6、12、24及48 h）的IL-12、CCL19及CCL22的表達水平明顯高于未負載組(P lt;0.01)。④K-ras負載組fascin-1蛋白的表達水平也高于未負載組(P lt;0.01)。
結論 K-ras突變多肽能夠促進DC成熟，并使細胞趨化因子和骨架蛋白表達水平增加，能夠增強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胞對CCL19、CCL22和fascin-1表達的影響. 中國普外基礎與臨床雜志, 2010, 17(9): 917-921. doi: 復制

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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.
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7. 薛剛， 曹永寬， 王培紅， 等. 腫瘤細胞裂解物致敏的樹突狀細胞對結腸癌LoVo細胞的殺傷作用 [J]. 中國普外基礎與臨床雜志， 2009； 16(1)： 4851.
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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)樹突狀細胞疫苗誘導抗胰腺癌免疫反應 [J]. 蘇州大學學報（醫(yī)學版）, 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胞誘導大鼠肝移植免疫低反應性的實驗研究 [J]. 中國普外基礎與臨床雜志, 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. 腫瘤細胞裂解物致敏的樹突狀細胞對結腸癌LoVo細胞的殺傷作用 [J]. 中國普外基礎與臨床雜志， 2009； 16(1)： 4851.
  8. 8. 譚廣， 任金帥， 王忠裕， 等. TNFα增強樹突狀細胞誘導的抗胰腺癌免疫應答的實驗研究 [J]. 中國普外基礎與臨床雜志， 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)樹突狀細胞疫苗誘導抗胰腺癌免疫反應 [J]. 蘇州大學學報（醫(yī)學版）, 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胞誘導大鼠肝移植免疫低反應性的實驗研究 [J]. 中國普外基礎與臨床雜志, 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.