• 1 南方醫(yī)科大學(xué)南方醫(yī)院內(nèi)分泌代謝科（廣州，510515）;2 吉安市中心人民醫(yī)院內(nèi)分泌科;

【摘要】　目的　 觀察晚期糖基化終產(chǎn)物（advanced glycosylation end prodrcts，AGE）對人結(jié)腸癌細(xì)胞株SW-480增殖的影響，并探討其可能機(jī)制。　方法　 不同濃度AGE干預(yù)SW-480細(xì)胞，噻唑藍(lán)（MTT）法比較各組細(xì)胞活力，流式細(xì)胞術(shù)觀察AGE對SW-480細(xì)胞周期的影響，蛋白質(zhì)印跡法觀察AGE對SW-480細(xì)胞CyclinD1表達(dá)的影響，端粒重復(fù)序列擴(kuò)增法（telomeric repeat amplification protocol，TRAP）銀染法觀察AGE對SW-480細(xì)胞端粒酶活性的影響。MTT測細(xì)胞活力的檢測設(shè)置空白對照組、100 μg/mL小牛血清白蛋白（bovine serum albumin，BSA）組及50、100、500 μg/mL AGE組，其余檢測只設(shè)置100 μg/mL BSA組和100 μg/mL AGE組。　結(jié)果　 MTT結(jié)果示AGE促進(jìn)SW-480細(xì)胞的增殖，且呈濃度依賴性。100 μg/mL BSA組與100 μg/mL AGE組72 h后的細(xì)胞G0/G1期所占百分比分別為56.02%±0.58%、51.93%±1.01%，差異有統(tǒng)計學(xué)意義（P lt;0.05）。蛋白質(zhì)印跡法示100 μg/mL AGE組72 h后CyclinD1的表達(dá)較100 μg/mL BSA組增加，差異有統(tǒng)計學(xué)意義（P lt;0.05）。TRAP銀染法檢測示100 μg/mL AGE干預(yù)SW-480細(xì)胞72 h后可以增加端粒酶活性（P lt;0.05）。　結(jié)論　 AGE可促進(jìn)人結(jié)腸癌細(xì)胞SW-480生長，呈劑量依賴性。其作用機(jī)制可能與AGE上調(diào)CyclinD1的表達(dá)加速G1/S期轉(zhuǎn)換及增加端粒酶活性有關(guān)。
【Abstract】　Objective　 To observe the effects of advanced glycosylation end products (AGE) on proliferation of SW-480 cells and study the possible mechanism.　Methods　 Various concentrations of AGE were designed to have impact on SW-480 cells. Proliferation of SW-480 cells was assessed by thiazolyl blue tetrazolium bromide (MTT) assay; The impact of AGE on the cell cycle of SW-480 cells was analyzed by flow cytometry (FCM); the influence of AGE on expression of CyclinD1 was checked by Western blotting; and the impact of AGE on telomerase activity was examined by telomeric repeat amplification proctol (TRAP) sliver staining. For the MTT assay, blank control group, 100 μg/mL bovine serum albumin (BSA) group, 50, 100 and 500 μg/mL AGE groups were designed, while for other examinations, there were only 100 μg/mL BSA group and 100 μg/mL AGE group.　Results　 MTT result showed that AGE increased the proliferation of SW-480 cells in a dose-dependent mode. The proportion of the cells at G0/G1 stage of the 100 μg/mL BSA group and the 100 μg/mL AGE experimental group were (56.02±0.58)% and (51.93±1.01)% respectively after 72 hours, with a significant difference (P lt;0.05); western blotting showed that the expression of CyclinD1 in the 100 μg/mL AGE group was significantly higher than that in the 100 μg/mL BSA group after 72 hours; TRAP silver staining demonstrated that telomerase activity increased significantly after treated with 100 μg/mL AGE for 72 hours.　Conclusions　 AGE can promote the growth of SW-480 cells in a dose-dependent mode. Its mechanism is mainly by up-regulating the expression of CyclinD1 to shorten G0/G1 and increasing the telomerase activity significantly.

引用本文： 周小智,薛耀明,關(guān)美萍,朱波. 晚期糖基化終產(chǎn)物對人結(jié)腸癌細(xì)胞SW-480增殖的影響及其機(jī)制研究. 華西醫(yī)學(xué), 2011, 26(6): 837-841. doi: 復(fù)制

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2. 　Inoue M, Iwasaki M, Otani T, et al. Diabetes mellitus and the risk of cancer: results from a large-scale population-based cohort study in Japan[J]. Arch Intern Med, 2006, 166(17): 1871-1877.
3. 　Vigneri P, Frasca F,Sciacca L, et al. Diabetes and cancer[J].Encocrine-Related Cancer, 2009,16(4): 1103-1123.
4. 　Edward F, Tierney MPH, Linda S. Population-based estimates of mortality associated with diabetes: use of a death certificate check box in North Dakota[J]. Am J Public Health, 2001, 91(1): 84-92.
5. 　Abe R, Shimizu T, Sugawara H, et al. Regulation of human melanoma growth and metastasis by AGE-AGE receptor interactions[J]. J Invest Dermatol, 2004, 122(2): 461-467.
6. 　Abe R, Yamagishi S. AGE-RAGE system and carcinogenesis[J]. Curr Pharm Des, 2008, 14(10): 940-945.
7. 　王愛萍, 胡水清, 湯哲, 等. 糖尿病與結(jié)直腸癌患病危險關(guān)系的調(diào)查分析[J]. 世界華人消化雜志, 2007, 15(1): 88-91.
8. 　Jullumstro E, Kollind M, Lydersen S, et al. Diabetes mellitus and outcomes of colorectal cancer[J]. Acta Oncol, 2009, 48(3): 361-367.
9. 　Berster JM, Goke B. Type 2 diabetes mellitus as risk factor for colorectal cancer[J]. Arch Physiol Biochem, 2008, 114(1): 84-98.
10. 　Larsson SC, Orsini N, Wolk A. Diabetes mellitus and risk of colorectal cancer: a meta-analysis[J]. J Natl Cancer Inst, 2005, 97(22): 1679-1687.
11. 　Yamagishi S, Nakamura K, Inoue H, et al. Possible participation of advanced glycation end products in the pathogenesis of colorectal cancer in diabetic patients[J]. Med Hypotheses, 2005, 64(6): 1208-1210.
12. 　王瑞良, 蘇青, 等.RAGE及其配體的臨床意義研究進(jìn)展[J]. 國際內(nèi)分泌代謝雜志, 2006, 26(3): 160-162.
13. 　Miki S, Kasayama S, Miki Y, et al. Expression of receptors for advanced glycosylation end products on renal cell carcinoma cells in vitro[J]. Biochem Biophys Res Commun, 1993, 196(2): 984-989.
14. 　Yamamoto Y, Yamagishi S, Hsu CC, et al. Advanced glycation end products-receptor interactions stimulate the growth of human pancreatic cancer cells through the induction of platelet-derived growth factor-B[J]. Biochem Biophys Res Commun, 1996, 222(3): 700-705.
15. 　Fuentes MK, Nigavekar SS, Arumugam T, et al. RAGE activation by S100P in colon cancer stimulates growth, migration and cell signaling pathway[J]. Dis Colon Rectum 2007, 50(8): 1230-1240.
16. 　Kuniyasu H, Chihara Y, Takahashi T. Co-expression of receptor for advanced glycation end products and the ligand amphoterin associates closely with metastasis of colorectal cancer [J]. Oncol Rep, 2003, 10(2): 445-448.
17. 　Ishiguro H, Nakaigawa N, Miyoshi Y, et al. Receptor for advanced glycation end products (RAGE) and its ligand, amphoterin are overexpressed and associated with prostate cancer development [J]. Prostate, 2005, 64(1): 92-100.
18. 　Guh JY, Huang JS, Chen HC, et al. Advanced glycation end product-induced proliferation in NRK-49F cells is dependent on the JAK2/STAT5 pathway and cyclin D1[J]. Am J Kidney Dis, 2001, 38(5): 1096-1104.
19. 　Kim JY, Park HK, Yoon JS, et al. Advanced glycation end product (AGE)-induced proliferation of HEL cells via receptor for AGE-related signal pathways[J]. Int J Oncol, 2008, 33(3): 493-501.
20. 　Zill H, Gunther R, Erbersdobler HF, et al. RAGE expression and AGE-induced MAP kinase activation in Caco-2 cells[J]. Biochem Biophys Res Commun, 2001, 288(5): 1108-1111.
21. 　Kuniyasu H, Chihara Y, Kondo H.Differential effects between amphoterin and advanced glycation end products on colon cancer cells[J]. Int J Cancer, 2003, 104(6): 722-727.
22. 　Boldrini L, Faviana P, Gisfredi S, et al. Evaluation of telomerase in the development and progression of colon cancer [J]. Int J Mol Med, 2002, 10(5): 589-592.
23. 　Holt SE, Wright WE, Shay JW. Multiple pathways for the regulation of telomerase activity [J]. Eur J Cancer, 1997, 33(5): 761-766.
  1. 1. 　Stattin P, Bjor O, Ferrari P, et al. Prospective study of hyperglycemia and cancer risk[J]. Diabetes Care, 2007, 30(3): 561-567.
  2. 2. 　Inoue M, Iwasaki M, Otani T, et al. Diabetes mellitus and the risk of cancer: results from a large-scale population-based cohort study in Japan[J]. Arch Intern Med, 2006, 166(17): 1871-1877.
  3. 3. 　Vigneri P, Frasca F,Sciacca L, et al. Diabetes and cancer[J].Encocrine-Related Cancer, 2009,16(4): 1103-1123.
  4. 4. 　Edward F, Tierney MPH, Linda S. Population-based estimates of mortality associated with diabetes: use of a death certificate check box in North Dakota[J]. Am J Public Health, 2001, 91(1): 84-92.
  5. 5. 　Abe R, Shimizu T, Sugawara H, et al. Regulation of human melanoma growth and metastasis by AGE-AGE receptor interactions[J]. J Invest Dermatol, 2004, 122(2): 461-467.
  6. 6. 　Abe R, Yamagishi S. AGE-RAGE system and carcinogenesis[J]. Curr Pharm Des, 2008, 14(10): 940-945.
  7. 7. 　王愛萍, 胡水清, 湯哲, 等. 糖尿病與結(jié)直腸癌患病危險關(guān)系的調(diào)查分析[J]. 世界華人消化雜志, 2007, 15(1): 88-91.
  8. 8. 　Jullumstro E, Kollind M, Lydersen S, et al. Diabetes mellitus and outcomes of colorectal cancer[J]. Acta Oncol, 2009, 48(3): 361-367.
  9. 9. 　Berster JM, Goke B. Type 2 diabetes mellitus as risk factor for colorectal cancer[J]. Arch Physiol Biochem, 2008, 114(1): 84-98.
  10. 10. 　Larsson SC, Orsini N, Wolk A. Diabetes mellitus and risk of colorectal cancer: a meta-analysis[J]. J Natl Cancer Inst, 2005, 97(22): 1679-1687.
  11. 11. 　Yamagishi S, Nakamura K, Inoue H, et al. Possible participation of advanced glycation end products in the pathogenesis of colorectal cancer in diabetic patients[J]. Med Hypotheses, 2005, 64(6): 1208-1210.
  12. 12. 　王瑞良, 蘇青, 等.RAGE及其配體的臨床意義研究進(jìn)展[J]. 國際內(nèi)分泌代謝雜志, 2006, 26(3): 160-162.
  13. 13. 　Miki S, Kasayama S, Miki Y, et al. Expression of receptors for advanced glycosylation end products on renal cell carcinoma cells in vitro[J]. Biochem Biophys Res Commun, 1993, 196(2): 984-989.
  14. 14. 　Yamamoto Y, Yamagishi S, Hsu CC, et al. Advanced glycation end products-receptor interactions stimulate the growth of human pancreatic cancer cells through the induction of platelet-derived growth factor-B[J]. Biochem Biophys Res Commun, 1996, 222(3): 700-705.
  15. 15. 　Fuentes MK, Nigavekar SS, Arumugam T, et al. RAGE activation by S100P in colon cancer stimulates growth, migration and cell signaling pathway[J]. Dis Colon Rectum 2007, 50(8): 1230-1240.
  16. 16. 　Kuniyasu H, Chihara Y, Takahashi T. Co-expression of receptor for advanced glycation end products and the ligand amphoterin associates closely with metastasis of colorectal cancer [J]. Oncol Rep, 2003, 10(2): 445-448.
  17. 17. 　Ishiguro H, Nakaigawa N, Miyoshi Y, et al. Receptor for advanced glycation end products (RAGE) and its ligand, amphoterin are overexpressed and associated with prostate cancer development [J]. Prostate, 2005, 64(1): 92-100.
  18. 18. 　Guh JY, Huang JS, Chen HC, et al. Advanced glycation end product-induced proliferation in NRK-49F cells is dependent on the JAK2/STAT5 pathway and cyclin D1[J]. Am J Kidney Dis, 2001, 38(5): 1096-1104.
  19. 19. 　Kim JY, Park HK, Yoon JS, et al. Advanced glycation end product (AGE)-induced proliferation of HEL cells via receptor for AGE-related signal pathways[J]. Int J Oncol, 2008, 33(3): 493-501.
  20. 20. 　Zill H, Gunther R, Erbersdobler HF, et al. RAGE expression and AGE-induced MAP kinase activation in Caco-2 cells[J]. Biochem Biophys Res Commun, 2001, 288(5): 1108-1111.
  21. 21. 　Kuniyasu H, Chihara Y, Kondo H.Differential effects between amphoterin and advanced glycation end products on colon cancer cells[J]. Int J Cancer, 2003, 104(6): 722-727.
  22. 22. 　Boldrini L, Faviana P, Gisfredi S, et al. Evaluation of telomerase in the development and progression of colon cancer [J]. Int J Mol Med, 2002, 10(5): 589-592.
  23. 23. 　Holt SE, Wright WE, Shay JW. Multiple pathways for the regulation of telomerase activity [J]. Eur J Cancer, 1997, 33(5): 761-766.