• 1.昆明醫(yī)學(xué)院第二附屬醫(yī)院肝膽二科(昆明 650101);;
  • 2.昆明醫(yī)學(xué)院(昆明 650031);

目的  探討血管生長(zhǎng)抑素基因聯(lián)合生長(zhǎng)抑素在體外對(duì)人胰腺癌細(xì)胞BXPC-3和血管內(nèi)皮細(xì)胞ECV-304增殖的抑制作用。方法  重組pcDNA3/angio質(zhì)粒轉(zhuǎn)染人胰腺癌細(xì)胞BXPC-3，利用RT-PCR和Western blot檢測(cè)其表達(dá)及分泌血管生長(zhǎng)抑素的情況； MTT法和流式細(xì)胞儀檢測(cè)生長(zhǎng)抑素和血管生長(zhǎng)抑素對(duì)BXPC-3和ECV-304細(xì)胞增殖的影響。結(jié)果  轉(zhuǎn)染后的BXPC-3細(xì)胞表達(dá)并分泌血管生長(zhǎng)抑素。一定濃度（≥10 μg/ml）的生長(zhǎng)抑素對(duì)BXPC-3細(xì)胞的增殖有明顯抑制作用（P＜0.01），并在一定濃度范圍內(nèi)呈劑量依賴(lài)性，同時(shí)能誘導(dǎo)BXPC-3細(xì)胞凋亡（P＜0.01）； 但其對(duì)ECV-304細(xì)胞的增殖無(wú)明顯影響（P gt;0.05）。血管生長(zhǎng)抑素對(duì)ECV-304細(xì)胞的增殖有明顯抑制作用（P＜0.01），并能誘導(dǎo)其凋亡（P＜0.01）； 但對(duì)BXPC-3細(xì)胞的增殖無(wú)明顯影響（P gt;0.05）。血管生長(zhǎng)抑素聯(lián)合生長(zhǎng)抑素對(duì)BXPC-3及ECV-304細(xì)胞的增殖均有明顯抑制作用（P＜0.01），也可誘導(dǎo)其凋亡（P＜0.01），但無(wú)協(xié)同增強(qiáng)效應(yīng)。結(jié)論  ①生長(zhǎng)抑素主要通過(guò)直接抑制胰腺癌細(xì)胞增殖、促進(jìn)其凋亡而發(fā)揮抗胰腺癌作用； 在體外其對(duì)血管內(nèi)皮細(xì)胞無(wú)抑制作用。②血管生長(zhǎng)抑素主要通過(guò)特異性地抑制胰腺癌血管內(nèi)皮細(xì)胞的增殖并誘導(dǎo)其凋亡，從而抑制新生毛細(xì)血管的生成，致胰腺癌細(xì)胞壞死和腫瘤消退。

引用本文： 朱紅,郭永章,楊鴻煒,鄒浩,王琨,黃松泉. 血管生長(zhǎng)抑素基因聯(lián)合生長(zhǎng)抑素治療人胰腺癌的體外研究. 中國(guó)普外基礎(chǔ)與臨床雜志, 2009, 16(6): 460-465. doi: 復(fù)制

1. Chi SL, Pizzo SV. Angiostatin is directly cytotoxic to tumor cells at low extracellular pH: a mechanism dependent on cell surface-associated ATP synthase[Ｊ]. Cancer Res, 2006; 66(2): 875-882.
2. 張延齡. 胰腺癌的基因治療研究 [Ｊ]. 臨床外科雜志, 2004; 12(4): 242-243.
3. Tirone TA, Wang XP, Templeton NS, et al. Cell-specific cytotoxicity of human pancreatic adenocarcinoma cells using rat insulin promoter thymidine kinase-directed gene therapy [Ｊ]. World J Surg, 2004; 28(8): 826-833.
4. Mkinen K, Loimas S, Wahlfors J, et al. Evaluation of herpes simplex thymidine kinase mediated gene therapy in experimental pancreatic cancer [Ｊ]. J Gene Med, 2000; 2(5): 361-367.
5. Carrió M, Visa J, Cascante A, et al. Intratumoral activation of cyclophosphamide by retroviral transfer of the cytochrome P450 2B1 in a pancreatic tumor model. Combination with the HSVtk/GCV system [Ｊ]. J Gene Med, 2002; 4(2):141-149.
6. Shen GH, Ghazizadeh M, Kawanami O, et al. Prognostic significance of vascular endothelial growth factor expression in human ovarian carcinoma[Ｊ]. Br J Cancer, 2000; 83(2):196-203.
7. Ikeda N, Nakajima Y, Sho M, et al. The association of K-ras gene mutation and vascular endothelial growth factor gene expression in pancreatic carcinoma[Ｊ]. Cancer, 2001; 92(3): 488-499.
8. 李繼坤， 鄭淼， 李遠(yuǎn)芳， 等. p53及VEGF表達(dá)與結(jié)、直腸癌血道轉(zhuǎn)移發(fā)生的關(guān)系 [Ｊ]. 中國(guó)普外基礎(chǔ)與臨床雜志， 2002； 9（2）： 112-114.
9. Benjamin LE, Keshet E. Conditional switching of vascular endothelial growth factor (VEGF) expression in tumors: induction of endothelial cell shedding and regression of hemangioblastoma-like vessels by VEGF withdrawal [Ｊ]. Proc Natl Acad Sci USA, 1997; 94(16): 8761-8766.
10. Gourley M, Williamson JS. Angiogenesis: new targets for the development of anticancer chemotherapies [Ｊ]. Curr Pharm Des, 2000; 6(4): 417-439.
11. Niedergethmann M, Hildenbrand R, Wostbrock B, et al. High expression of vascular endothelial growth factor predicts early recurrence and poor prognosis after curative resection for ductal adenocarcinoma of the pancreas [Ｊ]. Pancreas, 2002; 25(2): 122-129.
12. 劉江偉， 李開(kāi)宗， 竇科峰. 環(huán)氧化酶-2和血管內(nèi)皮生長(zhǎng)因子在胰腺癌組織中的表達(dá)及其相關(guān)性研究 [Ｊ]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2003; 10（6）: 539-541.
13. Galaup A, Opolon P, Bouquet C, et al. Combined effects of docetaxel and angiostatin gene therapy in prostate tumor model [Ｊ]. Mol Ther, 2003; 7(6): 731-740.
14. 何長(zhǎng)生， 全竹富. 生長(zhǎng)抑素抗腫瘤機(jī)制及其對(duì)胰腺癌的治療展望 [Ｊ]. 腸外與腸內(nèi)營(yíng)養(yǎng)， 2006; 13(6): 373-376.
15. Kouroumalis E, Skordilis P, Thermos K, et al. Treatment of hepatocellular carcinoma with octreotide: a randomised controlled study[Ｊ]. Gut, 1998; 42(3): 442-447.
16. Casini Raggi C, Calabrò A, Renzi D, et al. Quantitative evaluation of somatostatin receptor subtype 2 expression in sporadic colorectal tumor and in the corresponding normal mucosa [Ｊ]. Clin Cancer Res, 2002; 8(2): 419-427.
17. 華赟鵬， 賴(lài)佳明， 梁力建， 等. 奧曲肽下調(diào)cMet的表達(dá)抑制肝細(xì)胞癌的生長(zhǎng) [Ｊ]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2006; 13（2）: 158-162.
18. 張燕玲， 張正. 生長(zhǎng)抑素類(lèi)似物對(duì)肝癌細(xì)胞凋亡及c-myc蛋白表達(dá)的影響 [Ｊ]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2003; 10（1）： 25-27.
19. Radulovic S, Comaru-Schally AM, Milovanovic S, et al. Somatostatin analogue RC-160 and LH-RH antagonist SB-75 inhibit growth of MIA PaCa-2 human pancreatic cancer xenografts in nude mice [Ｊ]. Pancreas， 1993; 8(1): 88-97.
20. 劉正人， 吳高松， 杜志勇， 等. 奧曲肽對(duì)胰腺癌細(xì)胞轉(zhuǎn)染SSTR2后的作用 [Ｊ]. 中國(guó)普通外科雜志， 2004； 13(6)： 424-427.
21. Patel YC, Panetta R, Escher E, et al. Expression of multiple somatostatin receptor genes in AtT-20 cells. Evidence for a novel somatostatin-28 selective receptor subtype [Ｊ]. J Biol Chem， 1994; 269(2): 1506-1509.
22. 張浩, 李昱驥, 周建平, 等. 表皮生長(zhǎng)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(guó)普外基礎(chǔ)與臨床雜志, 2008; 15(3): 180-184.
23. 張群華， 倪泉興， 沈兆中， 等. 生長(zhǎng)抑素對(duì)胰腺癌生長(zhǎng)抑制和凋亡作用的實(shí)驗(yàn)研究 [Ｊ]. 外科理論與實(shí)踐， 2002； 7（5）：352-355.
24. Zalatnai A, Szegedi Z, Bocsi J. Flow cytometric evidence of apoptosis in human pancreatic cancer xenografts treated with Sandostatin (octreotide) [Ｊ]. Anticancer Res， 2000; 20(3A): 1663-1666.
25. Qin RY, Fang RL, Gupta MK, et al. Alteration of somatostatin receptor subtype 2 gene expression in pancreatic tumor angiogenesis [Ｊ]. World J Gastroenterol， 2004; 10(1): 132-135.
26. Kumar M, Liu ZR, Thapa L, et al. Antiangiogenic effect of somatostatin receptor subtype 2 on pancreatic cancer cell line: Inhibition of vascular endothelial growth factor and matrix metalloproteinase-2 expression in vitro [Ｊ]. World J Gastroenterol， 2004; 10(3): 393-399.
27. Ren SG, Ezzat S, Melmed S, et al. Somatostatin analog induces insulin-like growth factor binding protein-1 (IGFBP-1) expression in human hepatoma cells [Ｊ]. Endocrinology, 1992; 131(5): 2479-2481.
  1. 1. Chi SL, Pizzo SV. Angiostatin is directly cytotoxic to tumor cells at low extracellular pH: a mechanism dependent on cell surface-associated ATP synthase[Ｊ]. Cancer Res, 2006; 66(2): 875-882.
  2. 2. 張延齡. 胰腺癌的基因治療研究 [Ｊ]. 臨床外科雜志, 2004; 12(4): 242-243.
  3. 3. Tirone TA, Wang XP, Templeton NS, et al. Cell-specific cytotoxicity of human pancreatic adenocarcinoma cells using rat insulin promoter thymidine kinase-directed gene therapy [Ｊ]. World J Surg, 2004; 28(8): 826-833.
  4. 4. Mkinen K, Loimas S, Wahlfors J, et al. Evaluation of herpes simplex thymidine kinase mediated gene therapy in experimental pancreatic cancer [Ｊ]. J Gene Med, 2000; 2(5): 361-367.
  5. 5. Carrió M, Visa J, Cascante A, et al. Intratumoral activation of cyclophosphamide by retroviral transfer of the cytochrome P450 2B1 in a pancreatic tumor model. Combination with the HSVtk/GCV system [Ｊ]. J Gene Med, 2002; 4(2):141-149.
  6. 6. Shen GH, Ghazizadeh M, Kawanami O, et al. Prognostic significance of vascular endothelial growth factor expression in human ovarian carcinoma[Ｊ]. Br J Cancer, 2000; 83(2):196-203.
  7. 7. Ikeda N, Nakajima Y, Sho M, et al. The association of K-ras gene mutation and vascular endothelial growth factor gene expression in pancreatic carcinoma[Ｊ]. Cancer, 2001; 92(3): 488-499.
  8. 8. 李繼坤， 鄭淼， 李遠(yuǎn)芳， 等. p53及VEGF表達(dá)與結(jié)、直腸癌血道轉(zhuǎn)移發(fā)生的關(guān)系 [Ｊ]. 中國(guó)普外基礎(chǔ)與臨床雜志， 2002； 9（2）： 112-114.
  9. 9. Benjamin LE, Keshet E. Conditional switching of vascular endothelial growth factor (VEGF) expression in tumors: induction of endothelial cell shedding and regression of hemangioblastoma-like vessels by VEGF withdrawal [Ｊ]. Proc Natl Acad Sci USA, 1997; 94(16): 8761-8766.
  10. 10. Gourley M, Williamson JS. Angiogenesis: new targets for the development of anticancer chemotherapies [Ｊ]. Curr Pharm Des, 2000; 6(4): 417-439.
  11. 11. Niedergethmann M, Hildenbrand R, Wostbrock B, et al. High expression of vascular endothelial growth factor predicts early recurrence and poor prognosis after curative resection for ductal adenocarcinoma of the pancreas [Ｊ]. Pancreas, 2002; 25(2): 122-129.
  12. 12. 劉江偉， 李開(kāi)宗， 竇科峰. 環(huán)氧化酶-2和血管內(nèi)皮生長(zhǎng)因子在胰腺癌組織中的表達(dá)及其相關(guān)性研究 [Ｊ]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2003; 10（6）: 539-541.
  13. 13. Galaup A, Opolon P, Bouquet C, et al. Combined effects of docetaxel and angiostatin gene therapy in prostate tumor model [Ｊ]. Mol Ther, 2003; 7(6): 731-740.
  14. 14. 何長(zhǎng)生， 全竹富. 生長(zhǎng)抑素抗腫瘤機(jī)制及其對(duì)胰腺癌的治療展望 [Ｊ]. 腸外與腸內(nèi)營(yíng)養(yǎng)， 2006; 13(6): 373-376.
  15. 15. Kouroumalis E, Skordilis P, Thermos K, et al. Treatment of hepatocellular carcinoma with octreotide: a randomised controlled study[Ｊ]. Gut, 1998; 42(3): 442-447.
  16. 16. Casini Raggi C, Calabrò A, Renzi D, et al. Quantitative evaluation of somatostatin receptor subtype 2 expression in sporadic colorectal tumor and in the corresponding normal mucosa [Ｊ]. Clin Cancer Res, 2002; 8(2): 419-427.
  17. 17. 華赟鵬， 賴(lài)佳明， 梁力建， 等. 奧曲肽下調(diào)cMet的表達(dá)抑制肝細(xì)胞癌的生長(zhǎng) [Ｊ]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2006; 13（2）: 158-162.
  18. 18. 張燕玲， 張正. 生長(zhǎng)抑素類(lèi)似物對(duì)肝癌細(xì)胞凋亡及c-myc蛋白表達(dá)的影響 [Ｊ]. 中國(guó)普外基礎(chǔ)與臨床雜志, 2003; 10（1）： 25-27.
  19. 19. Radulovic S, Comaru-Schally AM, Milovanovic S, et al. Somatostatin analogue RC-160 and LH-RH antagonist SB-75 inhibit growth of MIA PaCa-2 human pancreatic cancer xenografts in nude mice [Ｊ]. Pancreas， 1993; 8(1): 88-97.
  20. 20. 劉正人， 吳高松， 杜志勇， 等. 奧曲肽對(duì)胰腺癌細(xì)胞轉(zhuǎn)染SSTR2后的作用 [Ｊ]. 中國(guó)普通外科雜志， 2004； 13(6)： 424-427.
  21. 21. Patel YC, Panetta R, Escher E, et al. Expression of multiple somatostatin receptor genes in AtT-20 cells. Evidence for a novel somatostatin-28 selective receptor subtype [Ｊ]. J Biol Chem， 1994; 269(2): 1506-1509.
  22. 22. 張浩, 李昱驥, 周建平, 等. 表皮生長(zhǎng)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(guó)普外基礎(chǔ)與臨床雜志, 2008; 15(3): 180-184.
  23. 23. 張群華， 倪泉興， 沈兆中， 等. 生長(zhǎng)抑素對(duì)胰腺癌生長(zhǎng)抑制和凋亡作用的實(shí)驗(yàn)研究 [Ｊ]. 外科理論與實(shí)踐， 2002； 7（5）：352-355.
  24. 24. Zalatnai A, Szegedi Z, Bocsi J. Flow cytometric evidence of apoptosis in human pancreatic cancer xenografts treated with Sandostatin (octreotide) [Ｊ]. Anticancer Res， 2000; 20(3A): 1663-1666.
  25. 25. Qin RY, Fang RL, Gupta MK, et al. Alteration of somatostatin receptor subtype 2 gene expression in pancreatic tumor angiogenesis [Ｊ]. World J Gastroenterol， 2004; 10(1): 132-135.
  26. 26. Kumar M, Liu ZR, Thapa L, et al. Antiangiogenic effect of somatostatin receptor subtype 2 on pancreatic cancer cell line: Inhibition of vascular endothelial growth factor and matrix metalloproteinase-2 expression in vitro [Ｊ]. World J Gastroenterol， 2004; 10(3): 393-399.
  27. 27. Ren SG, Ezzat S, Melmed S, et al. Somatostatin analog induces insulin-like growth factor binding protein-1 (IGFBP-1) expression in human hepatoma cells [Ｊ]. Endocrinology, 1992; 131(5): 2479-2481.