劉宇 1 , 張浩 2 , 楊野 1 , 王欣 1 , 李鐵民 1 , 郭仁宣 2
  • 1.中國醫(yī)科大學(xué)附屬第一醫(yī)院干診科（沈陽 110001）;;
  • 2.中國醫(yī)科大學(xué)附屬第一醫(yī)院普外科（沈陽 110001）;

目的  觀察表皮生長因子（EGF）對胰腺癌細(xì)胞增殖、黏附及侵襲力的影響及其對核因子-κB（NF-κB）和細(xì)胞基質(zhì)金屬蛋白酶（MMPs）表達(dá)的影響，探討EGF促進(jìn)胰腺癌侵襲的相關(guān)機(jī)理。
方法  通過細(xì)胞增殖、黏附及侵襲實(shí)驗(yàn)，觀察在EGF影響下胰腺癌細(xì)胞黏附、增殖及侵襲能力變化; 通過RT-PCR、MMPs明膠酶譜分析、Western blot及EMSA，檢測胰腺癌細(xì)胞的MMP-2和MMP-9 mRNA及其蛋白表達(dá)以及NF-κB活性變化; 并觀察NF-κB抑制物吡咯烷二硫代氨基甲酸鹽（PDTC）對EGF誘導(dǎo)的胰腺癌細(xì)胞NF-κB活性、MMP-9 mRNA及其蛋白表達(dá)以及腫瘤細(xì)胞侵襲力的變化。
結(jié)果  EGF能夠增強(qiáng)胰腺癌細(xì)胞的侵襲能力，具有劑量依賴性（P＜0.05）。EGF對胰腺癌細(xì)胞的黏附力及增殖力無明顯影響。EGF處理后，腫瘤細(xì)胞的MMP-9蛋白和mRNA表達(dá)明顯升高，EGF濃度為50 ng/ml時(shí)，腫瘤細(xì)胞的MMP-9表達(dá)最強(qiáng)，但對MMP-2蛋白和mRNA的表達(dá)則無影響。隨著EGF濃度的增加，NF-κB活性增強(qiáng)，具有濃度依賴性（P＜0.05）。與EGF單獨(dú)處理相比，經(jīng)PDTC和EGF共同處理后的胰腺癌細(xì)胞NF-κB活性與MMP-9蛋白和mRNA表達(dá)均降低; PDTC和EGF共同處理后的胰腺癌細(xì)胞侵襲力較EGF單獨(dú)處理和對照均減弱（P＜0.05）。
結(jié)論  EGF通過激活NF-κB的活性，誘導(dǎo)MMP-9表達(dá)，促進(jìn)胰腺癌細(xì)胞的侵襲，而這一過程可以被PDTC抑制。

引用本文： 劉宇,張浩,楊野,王欣,李鐵民,郭仁宣. 表皮生長因子介導(dǎo)的NF-κB促進(jìn)胰腺癌細(xì)胞MMP-9表達(dá)及侵襲的實(shí)驗(yàn)研究. 中國普外基礎(chǔ)與臨床雜志, 2009, 16(8): 603-608. doi: 復(fù)制

1. Farrow B, Sugiyama Y, Chen A, et al. Inflammatory mechanisms contributing to pancreatic cancer development [Ｊ]. Ann Surg, 2004; 239(6): 763-771.
2. 張浩, 李昱驥, 周建平, 等. 表皮生長因子促進(jìn)胰腺癌細(xì)胞侵襲和轉(zhuǎn)移的實(shí)驗(yàn)研究 [Ｊ]. 中國普外基礎(chǔ)與臨床雜志, 2008; 15(3): 180-184.
3. Felx M, Guyot MC, Isler M, et al. Endothelin-1 (ET-1) promotes MMP-2 and MMP-9 induction involving the transcription factor NF-κB in human osteosarcoma [Ｊ]. Clin Sci (Lond), 2006; 110(6): 645-654.
4. Katoh M, Katoh M. Transcriptional mechanisms of WNT5A based on NF-κB, Hedgehog, TGFβ, and Notch signaling cascades [Ｊ]. Int J Mol Med, 2009; 23(6): 763-769.
5. 陸穎影, 靖大道, 王興鵬. 表皮生長因子受體與胰腺癌的關(guān)系及以其為靶向治療的研究進(jìn)展 [Ｊ]. 胃腸病學(xué), 2007; 12(1): 53-55.
6. 陳宗靜, 童賽雄. 表皮生長因子受體家族在胰腺癌與壺腹癌中表達(dá)情況的研究現(xiàn)狀 [Ｊ]. 肝膽胰外科雜志, 2004; 16(1): 76-77.
7. 陸澄, 曾水林, 張郢華, 等. 胰腺癌組織中P21(WAF1)、轉(zhuǎn)化生長因子α、表皮生長因子受體表達(dá)與神經(jīng)浸潤轉(zhuǎn)移的關(guān)系 [Ｊ]. 南通大學(xué)學(xué)報(bào)（醫(yī)學(xué)版）, 2006; 26(1): 31-32.
8. Festuccia C, Angelucci A, Gravina GL, et al. Epidermal growth factor modulates prostate cancer cell invasiveness regulating urokinase-type plasminogen activator activity. EGF-receptor inhibition may prevent tumor cell dissemination [Ｊ]. Thromb Haemost, 2005; 93(5): 964-975.
9. Banerjee S, Sengupta K, Saxena NK, et al. Epidermal gr-owth factor induces WISP-2/CCN5 expression in estrogen receptor-positive breast tumor cells through multiple molecular cross-talks [Ｊ]. Mol Cancer Res, 2005; 3(3): 151-162.
10. Nakatsuji Y, Nishio Y, Tani N, et al. Epidermal growth factor enhances invasive activity of BeWo choriocarcinoma cells by inducing 2 integrin expression [Ｊ]. Endocr J, 2003; 50(6): 703-714.
11. Nagai S, Nakamura M, Yanai K, et al. Gli1 contributes to the invasiveness of pancreatic cancer through matrix metalloproteinase-9 activation [Ｊ]. Cancer Sci, 2008; 99(7): 1377-1384.
12. Uchima Y, Sawada T, Hirakawa K. Action of antiproteases on pancreatic cancer cells [Ｊ]. JOP， 2007; 8(4 Suppl): 479-487.
13. Ellenrieder V, Hendler SF, Ruhland C, et al. TGF-β-induced invasiveness of pancreatic cancer cells is mediated by matrix metalloproteinase-2 and the urokinase plasminogen activator system [Ｊ]. Int J Cancer, 2001; 93(2): 204-211.
14. 肖敏, 陳婕, 余曉云, 等. Angiopoietin-2, MMP-9和TIMP-1在胰腺癌中的表達(dá)及其與胰腺癌侵襲轉(zhuǎn)移的關(guān)系 [Ｊ]. 臨床消化病雜志, 2006; 18(4): 216-218.
15. Tamahashi U, Kumagai J, Takizawa T, et al. Expression and intracellular localization of matrix metalloproteinases in intraductal papillary mucinous neoplasms of the pancreas [Ｊ]. Virchows Arch, 2008; 453(1): 79-87.
16. 張克君, 李德春, 高焱明, 等. NF-κB、 MMP-9與肝細(xì)胞肝癌浸潤轉(zhuǎn)移的實(shí)驗(yàn)研究 [Ｊ]. 中華肝膽外科雜志, 2006; 12(10): 691-694.
17. 張東濤, 楊力, 郭新寧, 等. 核因子-κB和MMP-9與胃癌侵襲轉(zhuǎn)移的關(guān)系 [Ｊ]. 腫瘤防治雜志， 2004; 11(6): 566-568.
18. Rhee JW, Lee KW, Sohn,WJ, et al. Regulation of matrix metalloproteinase-9 gene expression and cell migration by NF-κB in response to CpG-oligodeoxynucleotides in RAW 264.7 cells [Ｊ]. Mol Immunol, 2007; 44(6): 1393-1400.
19. Sliva D, English D, Lyons D, et al. Protein kinase C induces motility of breast cancers by upregulating secretion of urokinase-type plasminogen activator through activation of AP-1 and NF-κB [Ｊ]. Biochem Biophys Res Commun, 2002; 290(1): 552-557.
20. Alaniz L, García M, Cabrera P, et al. Modulation of matrix metalloproteinase-9 activity by hyaluronan is dependent on NF-κB activity in lymphoma cell lines with dissimilar invasive behavior [Ｊ]. Biochem Biophys Res Commun, 2004; 324(2): 736-743.
21. Zhang H, Ma G, Dong M, et al. Epidermal growth factor promotes invasiveness of pancreatic cancer cells through NF-κB-mediated proteinase productions [Ｊ]. Pancreas, 2006; 32(1): 101-109.
22. 張克君, 高焱明, 楊金鏞, 等. IκB-α基因轉(zhuǎn)染HCC9204細(xì)胞對NF-κB和MMP-9表達(dá)的影響 [Ｊ]. 中國普外基礎(chǔ)與臨床雜志, 2007; 14(2): 185-187.
23. Watabe T, Yoshida K, Shindoh M, et al. The Ets-1 and Ets-2 transcription factors activate the promoters for invasion-associated urokinase and collagenase genes in response to epidermal growth factor [Ｊ]. Int J Cancer, 1998; 77(1): 128-137.
24. Hahne JC, Okuducu AF, Sahin A, et al. The transcription factor ETS-1: its role in tumour development and strategies for its inhibition [Ｊ]. Mini Rev Med Chem, 2008; 8(11): 1095-1105.
25. Kim S, Choi JH, Kim JB, et al. Berberine suppresses TNF-α-induced MMP-9 and cell invasion through inhibition of AP-1 activity in MDA-MB-231 human breast cancer cells [Ｊ]. Molecules, 2008; 13(12): 2975-2985.
  1. 1. Farrow B, Sugiyama Y, Chen A, et al. Inflammatory mechanisms contributing to pancreatic cancer development [Ｊ]. Ann Surg, 2004; 239(6): 763-771.
  2. 2. 張浩, 李昱驥, 周建平, 等. 表皮生長因子促進(jìn)胰腺癌細(xì)胞侵襲和轉(zhuǎn)移的實(shí)驗(yàn)研究 [Ｊ]. 中國普外基礎(chǔ)與臨床雜志, 2008; 15(3): 180-184.
  3. 3. Felx M, Guyot MC, Isler M, et al. Endothelin-1 (ET-1) promotes MMP-2 and MMP-9 induction involving the transcription factor NF-κB in human osteosarcoma [Ｊ]. Clin Sci (Lond), 2006; 110(6): 645-654.
  4. 4. Katoh M, Katoh M. Transcriptional mechanisms of WNT5A based on NF-κB, Hedgehog, TGFβ, and Notch signaling cascades [Ｊ]. Int J Mol Med, 2009; 23(6): 763-769.
  5. 5. 陸穎影, 靖大道, 王興鵬. 表皮生長因子受體與胰腺癌的關(guān)系及以其為靶向治療的研究進(jìn)展 [Ｊ]. 胃腸病學(xué), 2007; 12(1): 53-55.
  6. 6. 陳宗靜, 童賽雄. 表皮生長因子受體家族在胰腺癌與壺腹癌中表達(dá)情況的研究現(xiàn)狀 [Ｊ]. 肝膽胰外科雜志, 2004; 16(1): 76-77.
  7. 7. 陸澄, 曾水林, 張郢華, 等. 胰腺癌組織中P21(WAF1)、轉(zhuǎn)化生長因子α、表皮生長因子受體表達(dá)與神經(jīng)浸潤轉(zhuǎn)移的關(guān)系 [Ｊ]. 南通大學(xué)學(xué)報(bào)（醫(yī)學(xué)版）, 2006; 26(1): 31-32.
  8. 8. Festuccia C, Angelucci A, Gravina GL, et al. Epidermal growth factor modulates prostate cancer cell invasiveness regulating urokinase-type plasminogen activator activity. EGF-receptor inhibition may prevent tumor cell dissemination [Ｊ]. Thromb Haemost, 2005; 93(5): 964-975.
  9. 9. Banerjee S, Sengupta K, Saxena NK, et al. Epidermal gr-owth factor induces WISP-2/CCN5 expression in estrogen receptor-positive breast tumor cells through multiple molecular cross-talks [Ｊ]. Mol Cancer Res, 2005; 3(3): 151-162.
  10. 10. Nakatsuji Y, Nishio Y, Tani N, et al. Epidermal growth factor enhances invasive activity of BeWo choriocarcinoma cells by inducing 2 integrin expression [Ｊ]. Endocr J, 2003; 50(6): 703-714.
  11. 11. Nagai S, Nakamura M, Yanai K, et al. Gli1 contributes to the invasiveness of pancreatic cancer through matrix metalloproteinase-9 activation [Ｊ]. Cancer Sci, 2008; 99(7): 1377-1384.
  12. 12. Uchima Y, Sawada T, Hirakawa K. Action of antiproteases on pancreatic cancer cells [Ｊ]. JOP， 2007; 8(4 Suppl): 479-487.
  13. 13. Ellenrieder V, Hendler SF, Ruhland C, et al. TGF-β-induced invasiveness of pancreatic cancer cells is mediated by matrix metalloproteinase-2 and the urokinase plasminogen activator system [Ｊ]. Int J Cancer, 2001; 93(2): 204-211.
  14. 14. 肖敏, 陳婕, 余曉云, 等. Angiopoietin-2, MMP-9和TIMP-1在胰腺癌中的表達(dá)及其與胰腺癌侵襲轉(zhuǎn)移的關(guān)系 [Ｊ]. 臨床消化病雜志, 2006; 18(4): 216-218.
  15. 15. Tamahashi U, Kumagai J, Takizawa T, et al. Expression and intracellular localization of matrix metalloproteinases in intraductal papillary mucinous neoplasms of the pancreas [Ｊ]. Virchows Arch, 2008; 453(1): 79-87.
  16. 16. 張克君, 李德春, 高焱明, 等. NF-κB、 MMP-9與肝細(xì)胞肝癌浸潤轉(zhuǎn)移的實(shí)驗(yàn)研究 [Ｊ]. 中華肝膽外科雜志, 2006; 12(10): 691-694.
  17. 17. 張東濤, 楊力, 郭新寧, 等. 核因子-κB和MMP-9與胃癌侵襲轉(zhuǎn)移的關(guān)系 [Ｊ]. 腫瘤防治雜志， 2004; 11(6): 566-568.
  18. 18. Rhee JW, Lee KW, Sohn,WJ, et al. Regulation of matrix metalloproteinase-9 gene expression and cell migration by NF-κB in response to CpG-oligodeoxynucleotides in RAW 264.7 cells [Ｊ]. Mol Immunol, 2007; 44(6): 1393-1400.
  19. 19. Sliva D, English D, Lyons D, et al. Protein kinase C induces motility of breast cancers by upregulating secretion of urokinase-type plasminogen activator through activation of AP-1 and NF-κB [Ｊ]. Biochem Biophys Res Commun, 2002; 290(1): 552-557.
  20. 20. Alaniz L, García M, Cabrera P, et al. Modulation of matrix metalloproteinase-9 activity by hyaluronan is dependent on NF-κB activity in lymphoma cell lines with dissimilar invasive behavior [Ｊ]. Biochem Biophys Res Commun, 2004; 324(2): 736-743.
  21. 21. Zhang H, Ma G, Dong M, et al. Epidermal growth factor promotes invasiveness of pancreatic cancer cells through NF-κB-mediated proteinase productions [Ｊ]. Pancreas, 2006; 32(1): 101-109.
  22. 22. 張克君, 高焱明, 楊金鏞, 等. IκB-α基因轉(zhuǎn)染HCC9204細(xì)胞對NF-κB和MMP-9表達(dá)的影響 [Ｊ]. 中國普外基礎(chǔ)與臨床雜志, 2007; 14(2): 185-187.
  23. 23. Watabe T, Yoshida K, Shindoh M, et al. The Ets-1 and Ets-2 transcription factors activate the promoters for invasion-associated urokinase and collagenase genes in response to epidermal growth factor [Ｊ]. Int J Cancer, 1998; 77(1): 128-137.
  24. 24. Hahne JC, Okuducu AF, Sahin A, et al. The transcription factor ETS-1: its role in tumour development and strategies for its inhibition [Ｊ]. Mini Rev Med Chem, 2008; 8(11): 1095-1105.
  25. 25. Kim S, Choi JH, Kim JB, et al. Berberine suppresses TNF-α-induced MMP-9 and cell invasion through inhibition of AP-1 activity in MDA-MB-231 human breast cancer cells [Ｊ]. Molecules, 2008; 13(12): 2975-2985.