胰腺癌細胞在二維和三維培養系統中的生長特性研究△_《中國普外基礎與臨床雜志》

作者：

徐強 , 胡亞 , 廖泉 ,  趙玉沛

中國醫學科學院北京協和醫院基本外科（北京 100730）;

關鍵詞：

胰腺癌多細胞球樣體三維培養 Ⅰ型膠原細胞外基質膠生長曲線細胞周期

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目的探討胰腺癌細胞在二維平面培養和三維培養（Ⅰ型膠原和細胞外基質膠）中的生長特性。方法將3株胰腺癌細胞株SW1990、PCT和ASPC1分別采用上述3種培養方式進行培養，觀察細胞生長形態。采用CCK8法測定細胞生長曲線，采用乙醇固定碘化丙錠染色法檢測細胞周期分布。結果細胞在二維平面培養系統中呈單層貼壁生長; 在Ⅰ型膠原和細胞外基質膠中細胞形成多細胞球樣體（multicellular spheroid，MCS），其生長速度較二維平面培養中的細胞慢。在二維平面培養中生長的SW1990、PCT和ASPC1細胞的S期細胞的比例分別為(29.6±3.0)%、(33.6±2.1)%和(33.1±1.8)%，明顯高于在Ⅰ型膠原中培養4 d和8 d形成的MCS的S期細胞比例〔(18.2±5.1)%、(14.5±3.2)%和(24.7±2.6)%〕，P lt;0.05，而G2/M期的細胞比例差異沒有統計學意義（P gt;0.05）。在Ⅰ型膠原中培養4 d和8 d的SW1990和PCT細胞以及培養8 d的ASPC1細胞的G0/G1期細胞比例明顯高于其在二維平面培養中的細胞比例（P lt;0.05）。在Ⅰ型膠原中培養4 d的ASPC1細胞和SW1990細胞的S期細胞比例明顯高于其培養8 d的細胞比例（P lt;0.05）。結論不同的培養方式、培養介質對胰腺癌細胞的生長具有較大的影響。MCS三維培養系統能更好地模擬體內腫瘤細胞的生長狀況。

引用本文： 徐強,胡亞,廖泉,趙玉沛. 胰腺癌細胞在二維和三維培養系統中的生長特性研究△. 中國普外基礎與臨床雜志, 2011, 18(1): 33-38. doi: 復制

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14.	徐強, 廖泉, 趙玉沛. 多細胞球樣體的制備及其在腫瘤治療研究中的應用 [J]. 基礎醫學與臨床, 2010, 30(3): 320324.
15.	Santini MT, Rainaldi G, Indovina PL. Multicellular tumour spheroids in radiation biology [J]. Int J Radiat Biol, 1999, 75(7): 787799.
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19.	潘玉先, 鄭振華. 人大隱靜脈內皮細胞種植人工血管的實驗研究 [J]. 中國普外基礎與臨床雜志, 2000, 7(1): 1315.
20.	施森, 何延政, 劉勇, 等. 外周血來源內皮祖細胞三維血管新生模型的建立及其特性分析 [J]. 中國普外基礎與臨床雜志, 2009, 16(2): 119123.
21.	Gen M, Castro Kreder N, Bartenvan Rijbroek A, et al. Enhancement of effects of irradiation by gemcitabine in a glioblastoma cell line and cell line spheroids [J]. J Cancer Res Clin Oncol, 2004, 130(1): 4551.
22.	Muir CP, Adams MA, Graham CH. Nitric oxide attenuates resistance to doxorubicin in threedimensional aggregates of human breast carcinoma cells [J]. Breast Cancer Res Treat, 2006, 96(2): 169176.
23.	Chua YJ, Zalcberg JR. Pancreatic cancer-is the wall crumbling? [J]. Ann Oncol, 2008, 19(7): 12241230.
24.	Vulfovich M, RochaLima C. Novel advances in pancreatic cancer treatment [J]. Expert Rev Anticancer Ther, 2008, 8(6): 9931002.
25.	Boeck S, Heinemann V. The role of secondline chemotherapy after gemcitabine failure in patients with advanced pancreatic cancer [J]. Future Oncol, 2008, 4(1): 4150.
26.	Tempero M, Plunkett W, Ruiz Van Haperen V, et al. Randomized Phase Ⅱ comparison of doseintense gemcitabine: thirtyminute infusion and fixed dose rate infusion in patients with pancreatic adenocarcinoma [J]. J Clin Oncol, 2003, 21(18): 34023408.
27.	Louvet C, Labianca R, Hammel P, et al. Gemcitabine in combination with oxaliplatin compared with gemcitabine alone in locally advanced or metastatic pancreatic cancer: results of a GERCOR and GISCAD phase Ⅲ trial [J]. J Clin Oncol, 2005, 23(15): 35093516.

1. Abbott A. Cell culture: biolog’s new dimension [J]. Nature, 2003, 424(6951): 870872.
2. Matsuura T. Bioreactors for 3dimensional highdensity culture of human cells [J]. Hum Cell, 2006, 19(1): 1116.
3. Del Duca D, Werbowetski T, Del Maestro RF. Spheroid preparation from hanging drops: characterization of a model of brain tumor invasion [J]. J Neurooncol, 2004, 67(3): 295303.
4. Lee GY, Kenny PA, Lee EH, et al. Threedimensional culture models of normal and malignant breast epithelial cells [J]. Nat Methods, 2007, 4(4): 359365.
5. Lin RZ, Chang HY. Recent advances in threedimensional multicellular spheroid culture for biomedical research [J]. Biotechnol J, 2008, 3(910): 11721184.
6. 周濤, 胡亞, 廖泉, 等. 2種體外腫瘤細胞藥敏檢測方法的比較研究 [J]. 中國普外基礎與臨床雜志, 2008, 15(11): 825828.
7. 周濤, 胡亞, 廖泉, 等. 胰腺癌多細胞團簇的化療藥物敏感試驗 [J]. 中華胰腺病雜志, 2008, 8(6): 361364.
8. Mellor HR, Ferguson DJ, Callaghan R. A model of quiescent tumor microregions for evaluating multicellular resistance to chemotherapeutic drugs [J]. Br J Cancer, 2005, 93(3): 302309.
9. KunzSchughart LA, Freyer JP, Hofstaedter F, et al. The use of 3D cultures for highthroughput screening: the multicellular spheroid model [J]. J Biomol Screen, 2004, 9(4): 273285.
10. MuellerKlieser W. Tumor biology and experimental therapeutics [J]. Crit Rev Oncol Hematol, 2000, 36(23): 123139.
11. Friedrich J, Ebner R, KunzSchughart LA. Experimental antitumor therapy in 3D: spheroids-old hat or new challenge? [J]. Int J Radiat Biol, 2007, 83(1112): 849871.
12. 周方正, 伍鋼. 細胞偽足與腫瘤轉移研究進展 [J]. 臨床腫瘤學雜志, 2007, 12(1): 7779.
13. 張秀真, 吳澤志. 細胞偽足形成與微絲骨架研究進展 [J]. 國外醫學臨床生物化學與檢驗學分冊, 2005, 26(4): 213216.
14. 徐強, 廖泉, 趙玉沛. 多細胞球樣體的制備及其在腫瘤治療研究中的應用 [J]. 基礎醫學與臨床, 2010, 30(3): 320324.
15. Santini MT, Rainaldi G, Indovina PL. Multicellular tumour spheroids in radiation biology [J]. Int J Radiat Biol, 1999, 75(7): 787799.
16. Sminia P, Acker H, Eikesdal HP, et al. Oxygenation and response to irradiation of organotypic multicellular spheroids of human glioma [J]. Anticancer Res, 2003, 23(2B): 14611466.
17. Acker H, Carlsson J, MuellerKlieser W, et al. Comparative pO2 measurements in cell spheroids cultured with different techniques [J]. Br J Cancer, 1987, 56(3): 325327.
18. DomínguezMuoz JE著, 趙玉沛主譯. 臨床胰腺病學 [M]. 天津: 天津科技翻譯出版公司, 2007: 292293.
19. 潘玉先, 鄭振華. 人大隱靜脈內皮細胞種植人工血管的實驗研究 [J]. 中國普外基礎與臨床雜志, 2000, 7(1): 1315.
20. 施森, 何延政, 劉勇, 等. 外周血來源內皮祖細胞三維血管新生模型的建立及其特性分析 [J]. 中國普外基礎與臨床雜志, 2009, 16(2): 119123.
21. Gen M, Castro Kreder N, Bartenvan Rijbroek A, et al. Enhancement of effects of irradiation by gemcitabine in a glioblastoma cell line and cell line spheroids [J]. J Cancer Res Clin Oncol, 2004, 130(1): 4551.
22. Muir CP, Adams MA, Graham CH. Nitric oxide attenuates resistance to doxorubicin in threedimensional aggregates of human breast carcinoma cells [J]. Breast Cancer Res Treat, 2006, 96(2): 169176.
23. Chua YJ, Zalcberg JR. Pancreatic cancer-is the wall crumbling? [J]. Ann Oncol, 2008, 19(7): 12241230.
24. Vulfovich M, RochaLima C. Novel advances in pancreatic cancer treatment [J]. Expert Rev Anticancer Ther, 2008, 8(6): 9931002.
25. Boeck S, Heinemann V. The role of secondline chemotherapy after gemcitabine failure in patients with advanced pancreatic cancer [J]. Future Oncol, 2008, 4(1): 4150.
26. Tempero M, Plunkett W, Ruiz Van Haperen V, et al. Randomized Phase Ⅱ comparison of doseintense gemcitabine: thirtyminute infusion and fixed dose rate infusion in patients with pancreatic adenocarcinoma [J]. J Clin Oncol, 2003, 21(18): 34023408.
27. Louvet C, Labianca R, Hammel P, et al. Gemcitabine in combination with oxaliplatin compared with gemcitabine alone in locally advanced or metastatic pancreatic cancer: results of a GERCOR and GISCAD phase Ⅲ trial [J]. J Clin Oncol, 2005, 23(15): 35093516.

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胰腺癌細胞在二維和三維培養系統中的生長特性研究△

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《中國普外基礎與臨床雜志》

胰腺癌細胞在二維和三維培養系統中的生長特性研究△

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